Tropical Biomedicine

Epidemiology, clinical features and transmission of re-emerging arboviral infection chikungunya Balamurugan Shanmugaraj, Ashwini Malla, Sathishkumar Ramalingam Asian Pacific Journal of Tropical Biomedicine 2019 9(4):135-139 A number of re-emerging and emerging infectious diseases including chikungunya, West Nile, yellow fever, Zika, dengue, Japanese encephalitis, and others have increased in recent years, which threaten the public health across the globe. Chikungunya is a neglected re-emerging arboviral infection caused by chikungunya virus. Arboviral infections such as chikungunya, Zika and dengue have similar epidemiology, transmission cycles and clinical symptoms, which makes it difficult to diagnose these three infections. Moreover, there is no commercial vaccine or licensed therapy available for chikungunya infection, thus causing severe burden worldwide. Vector control may reduce the disease risk; however, this remains a challenge due to many factors including, but not limited to, evolution of insecticide resistance in mosquitoes, gaps in vector control tools, urbanization, environmental and demographic changes. Effective integrated vector control strategies and surveillance measures along with affordable vaccine development or anti-viral therapy are essential to control the infection. In this review, we discuss the epidemiology of mosquito-borne infection chikungunya which has re-emerged as an international concern in recent decades.

Keladi candik (Alocasia longiloba Miq.) petiole extracts promote wound healing in a full thickness excision wound model in rats Nurul Hazirah Che Hamzah, Arifullah Mohammed, KNS Sirajudeen, Mohd Asnizam Asari, Zulhazman Hamzah, Ibrahim Khalivulla Shaik Asian Pacific Journal of Tropical Biomedicine 2019 9(4):140-149 Objective: To investigate the wound-healing effect of Alocasia longiloba (A. longiloba) petiole extract on wounds in rats. Methods: Twenty-two male Sprague-dawley rats were randomly assigned to receive 10% solcoseryl gel, phosphate buffer saline, 50% ethanol, 95% ethanol and hexane extracts of A. longiloba at 1.5%, 3% and 6% doses, respectively. A full thicknesses wound (6 mm) was created on the dorsal of the rat; and all rats were applied with the extract solutions, 10% solcoseryl gel and phosphate buffer saline once a day topically until day 12. The wound was photographed on day 1, 6 and 12, and the percentage of wound contraction was calculated. On day 12, rats were sacrificed and histological examination of granulation tissue was carried out using haematoxylin & eosin and Masson’s Trichrome stain to determine the wound healing effect. Results: In this study, 6% of 50% and 95% ethanol extracts of A. longiloba showed 82.50% and 82.32% wound contraction, respectively, and were comparable with 10% solcoseryl gel (82.30%). Meanwhile, phosphate buffer saline treated group showed the lowest wound contraction (69.86%). Histological assessment of wound treated with 6% of 95% ethanol extract of A. longiloba showed distinct epidermal and dermal layer, higher proliferation of fibroblast and more angiogenesis with collagen compared to other wound treated groups. Conclusions: A. longiloba petiole extracts have a wound healing potential and 6% of 95% ethanol extract of A. longiloba is more effective. Further studies are required to understand the wound healing mechanism of action of the extract.

Falcaria vulgaris extract attenuates diabetes–induced kidney injury in rats Jalili Cyrus, Roshankhah Shiva, Salahshoor Mohammad Reza Asian Pacific Journal of Tropical Biomedicine 2019 9(4):150-157 Objective: To assess the effects of Falcaria vulgaris (F. vulgaris) as an antioxidant on damage to kidney of diabetic rats. Methods: Diabetic rats were established via streptozotocin (60 mg/kg). Various doses of F. vulgaris extracts (50, 100 and 150 mg/kg) and streptozotocin + F. vulgaris extracts were administered via intraperitoneal (i.p) injection to 48 rats (n=8 per group) for 28 d. Subsequently, ferric ion reducing antioxidant power (FRAP) of renal tissue, thiobarbituric acid reactive species, blood glucose concentrations, insulin, nitrite oxide, the weight of animals, glomeruli characteristics and kidney function were evaluated. Results: Compared with the control rats, diabetic rats showed significant increase in malondialdehyde, blood urea nitrogen, creatinine, blood glucose, nitrite oxide contents in renal tissues, and glomerular diameter. Furthermore, tissue FRAP level, body weight, number of glomeruli and plasma insulin were markedly reduced in diabetic rats when compared with the control group (P < 0.05). However, in all F. vulgaris and F. vulgaris + streptozotocin groups, malondialdehyde level, blood urea nitrogen, creatinine, glomerular diameter, nitrite oxide, and glucose levels were decreased significantly; meanwhile, tissue FRAP level, body weight, glomeruli number and insulin serum level were increased, compared to the control diabetic group (P < 0.05). Conclusions: F. vulgaris extract alleviates renal damage in diabetic rats.

Antioxidant compounds and capacities of Gac (Momordica cochinchinensis Spreng) fruits Ali Abdulqader, Faisal Ali, Amin Ismail, Norhaizan Mohd Esa Asian Pacific Journal of Tropical Biomedicine 2019 9(4):158-167 Objective: To identify and determine the composition of antioxidant compounds, and to evaluate the antioxidant abilities of Gac fruit parts (peel, pulp, seed and aril) grown in Malaysia. Methods: LC-MS/MS was used for identification of antioxidant compounds and UV-Vis for estimation of the contents of phenolics, flavonoids, and carotenoids. Lycopene and β -carotene were quantified using high-performance liquid chromatography. DPPH (2, 2-diphenyl-1- picrylhydrazyl) and ferric reducing antioxidant power assays were employed to evaluate antioxidant capacities. Results: Phytochemicals were found amongst all the fruit parts. Notably, significant amounts of carotenoids [(107.4 ± 4.5), (85.7 ± 4.4), (110.6 ± 2.1) mg/100 g dry weight (DW)], and relatively high levels of both phenolics [(27.3 ± 1.7), (28.9 ± 2.4), (30.8 ± 2.7) mg/100 g DW] and flavonoids [(38.1 ± 2.2), (8.8 ± 1.3), (24.5 ± 3.3) mg/100 g DW] were found in the fruit’s peel, pulp and aril, respectively. Seed part also showed a relatively high level of flavonoids [(18.1 ± 2.3) mg/100 g DW]. Lycopene and β -carotene were found to be significantly high (P < 0.05) in aril [(579.3 ± 22.7) and (621.0 ± 35.0) μg/g DW], followed by peel [(51.0 ± 7.5) and (210.0 ± 12.5) μg/g DW] and pulp [(37.6 ± 10.9) and (205.6 ± 22.1) μg/g DW)]. Antioxidant assays revealed that aril possessed the highest scavenging activity (IC50 = 865 μg/mL), while the peel possessed the highest ferric reducing power of 140 pmol FeSO4/μg. Conclusions: The current results demonstrate that Gac fruit grown in Malaysia is a rich source of phytochemicals, especially carotenoids, and possesses antioxidant activities. Thus, such findings suggest Gac fruit as a source of an antioxidant plant.

GC–MS analysis and anti–mosquito activities of Juniperus virginiana essential oil against Anopheles stephensi (Diptera: Culicidae) Azar Tahghighi, Naseh Maleki-Ravasan, Navid Dinparast Djadid, Hamzeh Alipour, Raziyeh Ahmadvand, Fateh Karimian, Saeed Yousefinejad Asian Pacific Journal of Tropical Biomedicine 2019 9(4):168-175 Objective: To investigate phytochemicals present in the essential oil from aerial parts of eastern red cedar, Juniperus virginiana (J. virginiana) L. (Cupressaceae) and to determine its killing and repellent activities against larvae, pupae, and adults of the Asian malaria mosquito, Anopheles stephensi (Diptera: Culicidae). Methods: J. virginiana essential oil was extracted by hydrodistillation, and its chemical composition was determined by gas chromatography-mass spectrometry. Seven different logarithmic concentrations of J. virginiana essential oils were used in larvicidal and pupicidal assays. J. virginiana essential oils-impregnated bed nets were applied in a designed animal module to test excito-repellent activity against adult mosquitoes. Results: Fourteen constituents corresponding to 99.98% of J. virginiana essential oils were identified. Five main components were terpinen-4-ol (25.21%), camphor (19.89%), E-3-hexen- 1-ol (13.30%), γ-terpinene (7.86%), and l-menthone (2.27%). The LC50 and LC90 values against larvae of the Anopheles stephensi were 11.693 and 66.140 ppm and for pupae were 9.640 and 40.976 ppm, respectively. In excito-repellency assay, J. virginiana essential oils-impregnated bed nets provided an average of 54.63% protection for guinea pig and 45.37% mortality for the mosquitoes. Conclusions: Four monoterpenes and one leaf alcohol were identified by gas chromatography-mass spectrometry. J. virginiana essential oils showed potent larvicidal, pupicidal, adulticidal, and repellent activities against Anopheles stephensi at acceptable concentrations. Evaluation of bioactivity of identified chemicals (alone or in combination) will provide new eco-friendly substances for mosquito-management programs.

Antimalarial activities of butanol and ethylacetate fractions of Combretum nigricans leaf Enegide Chinedu, Peter A Akah, Dabum L Jacob, Ifeoma A Onah, Chimere Y Ukegbu, Chukwuma K Chukwuemeka Asian Pacific Journal of Tropical Biomedicine 2019 9(4):176-180 Objective: To evaluate the antimalarial activity of the ethylacetate and butanol fractions of Combretum nigricans (C. nigricans) leaf extract in mice. Methods: C. nigricans solvent (butanol and ethylacetate) fractions were screened for their phytochemical constituents using standard procedures illustrated by Harborne and Evans. The Peters’ 4-day suppressive test against early malaria infection, Rane’s curative test against established malaria and prophylactic test for residual activity were employed for evaluating the antimalarial potential in mice. Results: The phytochemical screening revealed the presence of alkaloids, terpenoids, saponins, and flavonoids in both fractions at different intensity. Both fractions exhibited significant antimalarial activity in all test models (P<0.05). The ethylacetate fraction of C. nigricans had better chemosuppressive and curative effects compared to the butanol fraction, which however, elicited a better chemoprophylactic effect. The chemosuppressive effect of C. nigricans ethylacetate fraction (200-800 mg/kg) was 77.6%, 69.1% and 86.1%; curative effect was 62.3%, 71.3% and 72.4%; while the chemoprophylactic activity was 32.1%, 48.6% and 61.2% respectively. C. nigricans butanol fraction (200-800 mg/kg) had 40.3%, 54.1% and 69.1% chemosuppression; 26.2%, 36.9% and 34.5% curative effect; and 48.4%, 70.0% and 87.4% chemoprophylaxis. Conclusions: Both solvent fractions of C. nigricans possess antimalarial activity, and may be useful at different stages of malaria therapy.

Lungs

Endobronchial metastasis: The challenge continues Neeraj Gupta Lung India 2019 36(3):181-182

New combined assessment of chronic obstructive pulmonary disease: Utilization, pitfalls, and association with spirometry Ruchi Dua, Ranjeeta Kumari, Vivek Yadav, Mayur Ranjan, Subodh Kumar, Mayank Mishra, Suryakant Tripathi Lung India 2019 36(3):183-187 Introduction: Classification of chronic obstructive pulmonary (COPD) disease has changed from being solely based on spirometric variables to combined assessment including symptom scores and history of exacerbations/ hospitalizations. There is both lack of awareness regarding change in its assessment as well as underutilization due to time constraints and seeming complexity. Moreover, treatment of COPD needs to be tailored according to the new combined assessment. Aims: Current study was planned to look at current stratification of patients according to new revised combined assessment (Global Initiative for Chronic Obstructive Lung Disease GOLD 2017) in comparison to old(GOLD 2011) as well as its incorporation in clinical practice. Co-relation between revised combined assessment and spirometric staging was also assessed. Methods: 418 consecutive COPD patients were enrolled, their dyspnea scores in terms of modified medical research council scale (mMRC), preceding history of hospitalization/ exacerbation over preceding one year and spirometric variables were recorded. Their stratification according to old and new classification recorded. Their past treatment records were reviewed and combined assessment if done recorded. Results: Substantial shift of categories is seen from C and D respectively to stage A and B on applying the new classification compared to old i.e more severe to less severe. Secondly, revised combined assessment is still highly underutilized. Revised combined assessment has positive co-relation with spirometry and post bronchodilator forced expiratory volume in 1 second(FEV1). Conclusions: Management of substantial number of stable COPD patients may need to be stepped down in accordance with revised combined assessment. There is a need to disseminate information regarding change in COPD classification and stress on its incorporation in our day-to day clinical practice. Revised combined assessment has positive co-relation with spirometry, stressing its utility even in peripheral centers without spirometry facilities.

Exacerbation of chronic obstructive pulmonary diseases as a risk factor of the skeletal muscle dysfunction Vitalii Poberezhets, Yuriy Mostovoy, Hanna Demchuk Lung India 2019 36(3):188-192 Background: Exacerbation of Chronic Obstructive Pulmonary Disease (COPD) contributes to increased systemic inflammation, oxidative stress, hypoxia, hypercapnia and other risk factors of the skeletal muscle dysfunction. Aims and Objectives: This study aimed to determine whether the frequency of the COPD exacerbations promotes deterioration of the skeletal muscle dysfunction in patients with COPD and to figure out which characteristics of the muscles will reflect this changes. Materials and Methods: We examined 98 male COPD patients, mean age 60.4±11.2 years, GOLD groups B, C, D. To assess the degree of skeletal muscle dysfunction we used hand-grip strength, 6-minute walk test, bioelectrical impedance analysis. Quality of life was evaluated using St. George's respiratory questionnaire (SGRQ). Results: Increase in the frequency of COPD exacerbations was associated with the decrease in the 6-minute walk test distance (r= -0.214, P = 0.034) and with the rise of sarcopenia according to the results of bioelectrical impedance analysis – lower fat-free mass index (r = -0.201, P = 0.047). Frequency of COPD exacerbations had a weak correlation with the degree of the activity limitation component of SGRQ (r = 0.436, P < 0.001). Conclusions: COPD exacerbation plays an important role in progression of the skeletal muscle dysfunction. It decreases endurance of the skeletal muscles, diminishing their size, which manifests itself in reduced exercise tolerance. Increase in the frequency of the COPD exacerbations also worsens all components of the life quality.

Nonadherence to treatment and quality of life among patients with chronic obstructive pulmonary disease Shorouk Mohsen, Fadia Zaki Hanafy, Aya Ahmed Fathy, Abdel-Hady El-Gilany Lung India 2019 36(3):193-198 Objectives: Chronic obstructive pulmonary disease (COPD) is one of the most common respiratory diseases. Assessment of medication nonadherence and quality of life (QOL) is important in such chronic diseases. The aim of the study was to describe treatment nonadherence and QOL of COPD patients and their associated factors. Materials and Methods: A cross-sectional study was performed in Mansoura Chest Hospital on 133 already diagnosed inpatient cases of COPD from March to August 2017. Data were collected using two validated questionnaires, Morisky Medication Adherence Scale to assess medication adherence and St George's Respiratory Questionnaire for COPD patients (SGRQ-c) to assess QOL. Results: About 45% of studied group were nonadherent to prescribed medications. Logistic regression analysis showed that duration of disease is the only independent risk factor for nonadherence. The scores on SGRQ-c indicated sever QOL affection in COPD. SGRQ-c scores show significant impairment with the presence of comorbidities, hospital admission, and frequency of exacerbations. Conclusions: Nonadherence to treatment was affected mainly by the duration of disease. QOL impairment in COPD is influenced by broad range of factors suggesting the importance of QOL assessment in those patients. No association was found between nonadherence to treatment and QOL.

Flexible bronchoscopy-guided vocal cord biopsy under conscious sedation: An option to surgical biopsy in special situations Ravindra M Mehta, Abhinav Singla, Pavankumar Biraris, Sheelu Srinivas, Lakshmipriya Srinivasan Lung India 2019 36(3):199-201 Introduction: Vocal cord (VC) biopsies are usually performed with rigid laryngoscopy under general anesthesia (GA). In patients with comorbidities such as cardiovascular or pulmonary disease, the risk of GA is high, with morbidity and potential mortality. We describe VC biopsy (VCB) in such a high-risk cohort, done safely and successfully with flexible bronchoscopy-VCB (FB-VCB) and conscious sedation. Methods: FB-VCB was done in two groups of patients – the first at high risk for GA due to comorbidities and the second group where VC lesions were found incidentally in the course of FB done for other reasons. FB was done with local anesthesia and conscious sedation, and the VC lesion was identified. Flexible forceps were introduced through the working channel of the bronchoscope, and the lesions were carefully sampled. Results: FB-VCB was performed in 15 patients (14 males and 1 female) with a mean age of 60.7 ± 12.1 years. Of these 15 patients, 6 patients were poor candidates for GA due to comorbidities and 9 patients had incidental VC lesions found during bronchoscopy. A diagnosis was made in 14/15 (93.3%) patients. Complications included a mild ooze and hoarseness of voice for a few days, which did not require any intervention. Conclusion: We report one of the few series of patients with VC biopsies done with FB (FB-VCB) under conscious sedation, without significant complications. It is safe and useful in high-risk patients who are poor candidates for GA, with a good diagnostic yield.

A single-center experience of pediatric foreign-body aspiration: A retrospective 4-year case series Mustafa Erman Dorterler, Osman Hakan Kocaman, Tansel Gunendi, Mehmet Emin Boleken Lung India 2019 36(3):202-206 Introduction: Foreign body aspirations (FBA) in children are serious life-threatening clinical conditions that require immediate intervention. In this study, it was aimed to retrospectively investigate the demographic features, clinical diagnosis and treatment methods of children admitted to our clinic due to FBA. Materials and Methods: The study included 86 children aged <16 years, diagnosed with tracheobronchial foreign body aspiration (FBA) between January 2013 and December 2017. All patients with two-way chest radiography were examined for foreign body aspiration diagnosis. In case of suspicion of diagnosis, low-dose multi-slice chest CT was taken. In cases of FBA diagnosis, rigid bronchoscopy was performed under sevoflurane and propofol anaesthesia supported by controlled ventilation. Evaluation was made of the patient demographic characteristics, type and localization of the foreign body removed with bronchoscopy and operation-related complications. Results: The mean age of the patients with FBA diagnosis was 3.17 years and 55.8% (n = 48) of the patients were male. The most commonly aspirated foreign body was nuts (peanut and hazelnut) (70%) and the most common finding on the chest radiographs was obstructive emphysema, determined on 51% of the patients. Bronchoscopy revealed that the foreign body was in the right main bronchus and left main bronchus in 41%. Conclusion: The main treatment for FBA is prevention. However, in patients applied with bronchoscopy for FBA, controlled ventilation and appropriate general anesthesia should be generally used. Early bronchoscopic intervention with safe anesthesia and controlled ventilation support will improve the success rates in FBA cases.

Patient characteristics and outcomes of a home mechanical ventilation program in a developing country Narongkorn Saiphoklang, Apichart Kanitsap, Pitchayapa Ruchiwit, Pattarin Pirompanich, Thiti Sricharoenchai, Christopher B Cooper Lung India 2019 36(3):207-211 Background: There are limited data on home mechanical ventilation (HMV) in developing countries. This study aimed to describe the patient characteristics, feasibility, and outcomes of an HMV program at a university hospital in Thailand. Materials and Methods: Data were collected on all patients who were discharged with HMV between October 2014 and August 2015 at Thammasat University Hospital. Results: Twelve patients (eight men and four women) underwent HMV. They were aged 71.5 ± 17.6 years; mean ± standard deviation. Indications for HMV were 6 neurologic diseases (4 amyotrophic lateral sclerosis, 1 multiple system atrophy, and 1 stroke), 2 chronic obstructive pulmonary disease (COPD), 1 tracheomalacia, and 3 combined neurologic diseases and respiratory diseases (2 stroke and COPD, 1 stroke and tracheomalacia). The duration of follow-up was 799.5 ± 780.5 days. The ratio of family income to cost of HMV usage was 77.2:1 ± 5.5:1. All patients had tracheostomies. Modes of HMV were biphasic positive airway pressure (66.7%), pressure-controlled ventilation (16.7%), pressure-support ventilation (8.3%), and volume-controlled ventilation (8.3%). Complications occurred in ten patients (83.3%), including tracheobronchitis (20 events) and ventilator-associated pneumonia (12 events). Overall mortality was 41.7% (5/12 patients), including two patients who died due to ventilator-associated pneumonia. There were no instances of ventilator malfunction. Conclusions: HMV is feasible for patients with neurological diseases and COPD in a developing country. The relatively high rate of complications indicates the need for more comprehensive clinical services for chronic ventilator-dependent patients in this setting.

Endobronchial metastasis from extrathoracic malignancies: A clinicopathological study of 11 cases Monika Breta, Sudheer Arava, Karan Madan, Ashok Singh, Deepali Jain, Randeep Guleria Lung India 2019 36(3):212-215 Introduction: Endobronchial metastases are uncommon. It accounts for 2-28%. Most common tumors that metastasis to the endobronchial site are breast, renal, endometrial and colon carcinomas. They have to be identified properly because they present poor prognosis and different treatment modalities when compared to that of primary lung carcinomas. Materials and Methods: We studied a total of 11 retrospective cases of Endobronchial metastasis. Detailed clinic pathological and radiological findings were analyzed along with a detailed Immunohistochemical workup and Endobronchial findings. Results: Mean age of presentation was 55 years with female predominance (Male to female ratio was 1:1.2). Most common tumors encountered were carcinoma breast (3), colorectal carcinomas (2), renal cell carcinoma (2) followed by one each from carcinoma endometrium, cervix and thyroid. Common symptom was cough followed by dyspnea. Exophytic endobronchial growth was common with right main bronchus being the common site. Conclusion: Endobronchial metastasis occurs in various types of malignancies. It should always be differentiated from primary lung carcinomas as they carries poor prognosis and different treatment protocol when compared to that of primary lung tumors.

Pneumococcal disease burden from an Indian perspective: Need for its prevention in pulmonology practice Parvaiz A Koul, Sudhir Chaudhari, Ramesh Chokhani, D Christopher, Raja Dhar, Kumar Doshi, A Ghoshal, SK Luhadiya, Ashok Mahashur, Ravindra Mehta, Amita Nene, Md Rahman, Rajesh Swarnakar Lung India 2019 36(3):216-225 Globally, pneumococcal diseases are a significant public health concern. They are preventable and frequently occur among older adults. Major risk factors for the disease are extremes of age, alcohol intake, smoking, air pollution, and comorbid conditions (diabetes, chronic obstructive pulmonary disease, chronic kidney disease, liver disease, and heart disease). Risk factors, coupled with limited disease-burden data and the emergence of antibiotics resistance, are hindering the effective management of the disease in older adults. Various global guidelines recommend pneumococcal vaccines for the prevention of pneumococcal diseases, as they reduce disease burden, hospitalization, and mortality rates among patients with comorbid conditions. Besides being an integral part of childhood immunization, these vaccines are advocated by various Indian healthcare bodies/groups for older and younger adults with certain medical conditions. The article presents an overview of the closed-door discussion by the Indian pulmonary experts on the scientific evidence and clinical practice followed for the prevention of pneumococcal disease in India.

Barber's neck manipulation causing bilateral diaphragmatic paralysis and type-2 respiratory failure Ashish Kumar Prakash, Anand Jaiswal, Sandeep Mittal, Poulomi Chatterjee, Sameer Kotalwar, Bornalli Datta Lung India 2019 36(3):226-228 This is a case report of an unusual cause of bilateral diaphragmatic palsy. A 54-year-old gentleman, presented to us with exertional dyspnea and chest heaviness for the past 6 months which had increased in the last 6 days. Dyspnea increased on lying down. He was diagnosed as pneumonia on the basis of X-ray and chest CT scan, received treatment for the same and responded to the therapy. However, breathlessness and hypercapnia persisted. He had unexplained hypercapnia for which extensive investigations were carried out. Neurological and cardiac assessments were essentially normal. On revisit clinical examination, he was found to have paradoxical diaphragmatic movement with respiration. Ultrasound of chest detected no diaphragmatic movement. Detailed history elicited that patient was fond of neck massage and neck cracking wherein his barber would bend his neck with jerk to either side after a haircut.After considering all possible etiologies; we concluded that it was a case of diaphragm palsy induced by barber neck manipulation, leading to Type-2 respiratory failure. The fact that the vital clues to the diagnosis were elicited by detailed history and thorough examination reinforces that history and clinical examination for doctors shall remain a very important tool for clinical diagnosis.

Biomedical Research

Klotho: An emerging factor in neurodegenerative diseases Gauri V Pathare, Kavita K Shalia Biomedical Research Journal 2019 6(1):1-6 Soluble Klotho protein is present in blood, urine, and cerebrospinal fluid and works as a humoral factor exerting different biological effects. Several animal studies have demonstrated the association of age-related neurodegeneration with Klotho deficiency. Lower Klotho levels have been reported in patients suffering from cognitive impairment, dementia, Alzheimer's disease, Parkinson's disease, multiple sclerosis, and other neurodegenerative diseases. Due to its antiaging properties, Klotho is the obvious choice to be studied as a protective/therapeutic agent in neurobiology. In this review, we have attempted to shed light on the different neurodegenerative diseases affected by deficiency of Klotho and its neuroprotective role against pathogenicity of the disease.

Nitrate stress-induced bioactive sulfated polysaccharides from Chlamydomonas reinhardtii Jyoti Vishwakarma, Vaishnavi Parmar, Sirisha L Vavilala Biomedical Research Journal 2019 6(1):7-16 Sulfated polysaccharides (SPs) are anionic carbohydrate polymers synthesized as extracellular or cell wall components by most of the algae and have potent bioactive properties. In the current study, Chlamydomonas reinhardtii (Cr) cells were attributed to sodium nitrate stress in concentrations such as 5 mM, 10 mM, 20 mM, 30 mM, and a control to determine the productivity and bioactivity of SPs. SPs are extracted by hot water method using 80% ethanol. The percentage yield of SPs increased with an increase in concentration of sodium nitrate as compared to control. Biochemical analysis of the extract showed an increase in carbohydrate content (22%–95%), uronic acid content (23%–60%), and sulfate content from control to 30 mM NaNO3-treated extracts. The amount of reducing and nonreducing sugars was found to be 6.16% and 89.06%, respectively, while the protein content is ~16%. The antioxidant potential of SPs showed increased antioxidant activity with an increase in concentration of NaNO3 stress. The analysis resulted in maximum chelating activity of 83.73% assayed in concentration range of 1–8 μg/ml, total antioxidant activity of 70.36% in concentration 0.05–2μg/ml, and hydroxyl radical scavenging activity of 79.52% in concentration 250–1000 μg/ml; reducing potential was observed with the highest absorbance of 0.87; the 2,2-diphenyl-1-picrylhydrazyl scavenging activity showed the highest activity of 63.61%, while the superoxide scavenging activity was 92% at 0.1–1 μg/ml. Furthermore, Cr-SPs inhibited the growth of Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli bacterial growth as indicated by clear zones that increased in size with an increasing concentration of NaNO3. These results provide opportunities to develop Cr-SPs as natural antioxidant and antibacterial agents.

Molecular docking study for evaluation of neuroprotective potential of sericin against cerebral stroke and exploring its biomaterial properties Khushboo Maurya, Anand Kumar Pandey Biomedical Research Journal 2019 6(1):17-24 Background: Cerebral stroke, the third leading cause of death worldwide results from the improper blood supply to the brain due to occlusions in the brain arteries. This leads to production of free radicals contributed by cyclo-oxygenases (COX), acid sensing ion channels (ASIC) and matrix metalloproteinases (MMPs) causing adverse conditions of inflammation, oxidative stress, and acidosis leading to neuronal death thereby proving these enzymes as potent targets. Sericin, a 38 amino acid long protein found in silk fiber is known for its anti-inflammatory and anti-oxidant property. Aim and Objectives: Inhibition of the above-mentioned targets by silk protein sericin to reduce the pathological features by structural interactions as well as reducing inflammation and oxidative stress due to the natural properties of compound. Methodology: In the present study we studied structural inhibition of effective targets by sericin through molecular docking analysis. Also, the semi crystalline nature of sericin was deduced through in silico XRD spectral analysis. Result: Structural inhibition through molecular docking analysis proved highly efficient inhibition. Also, the in silico XRD spectral analysis proved sericin to be a potential biomaterial for scaffold development. Conclusion: Sericin can not only act as an effective drug against cerebral ischemia but can also be used to develop scaffold to repair damaged brain.

Transforming growth factor beta receptor 2 single-nucleotide polymorphism association with oral cancer and In silico identification of small drug-like molecules as inhibitors to transforming growth factor Beta-2 receptor Shaleen Multani, Hetal Damani Shah, Dhananjaya Saranath Biomedical Research Journal 2019 6(1):25-33 Objective: Oral cancer, in India, constitutes 26% of global oral cancer burden. The major risk factors include tobacco, areca nut, alcohol, and human papillomavirus 16/18; however, only 5%–10% of the high-risk individuals develop oral cancer, indicating the role of genomic variants in susceptibility to oral cancer. Conventional treatment options in oral cancer have resulted in relatively poor prognosis and an unmet need of treatment. In silico analysis, therefore, was performed to identify small drug-like molecules as potential inhibitors of transforming growth factor beta-2 receptor (TGFβRII). Materials and Methods: Seven single-nucleotide polymorphisms (SNPs) were analyzed in 500 histopathologically confirmed oral cancer samples and 500 long-term tobacco users (LTTUs) as controls using allelic discrimination real-time polymerase chain reaction or high-resolution melting analysis. The differential frequencies in oral cancer and LTTUs were calculated using SPSS software (version 19), and odds ratio (OR) to indicate risk to oral cancer using Hutchon.net. structure-based virtual screening of drug-like molecules was performed to identify lead inhibitor molecules to TGFβRII using Schrödinger Suite 2015-4. Results: Heterozygous GC genotype of TGFBR2 rs9843143 demonstrated increased risk ([P = 0.011; OR 1.61 [1.25–2.1]) while CC genotype showed decreased risk (P = 0.005; OR 0.61 [0.44–0.83]) to oral cancer. Increased/decreased risk to oral cancer was not observed for the other SNPs. In silico analysis identified six molecules as inhibitors of TGFβRII kinase domain from 17,723 conformers from Maybridge HitFinder library and 2685 conformers from MEGx AnalytiCon natural product library. Conclusion: SNP rs9843143 (TGFBR2) demonstrated a significant association (P < 0.05) with oral cancer and six potential inhibitors of TGFβRII kinase were identified using in silico analysis.

Metastatic hepatocellular carcinoma manifesting as primary esophageal carcinoma: A rare case report K Jayaprakash Shetty, HL Kishan Prasad, Shubha Bhat, Michelle Mathias, Vijith Shetty Biomedical Research Journal 2019 6(1):34-36 Hepatocellular carcinoma (HCC) metastasizing to the esophagus and mimicking as a primary tumor of the esophagus is extremely rare, being present in <0.4% in some autopsy series. This report describes a case of 70-year-old male with metastatic HCC to esophagogastric junction causing diagnostic dilemma. An endoscopic examination revealed an ulcerative lesion in the lower end of the esophagus. The biopsy specimen obtained from a tumor revealed the pseudoglandular arrangement of tumor cells. Ultrasound abdomen showed liver nodule with biopsy confirming as HCC. Immunohistochemistry (IHC) of the esophageal mass showed positivity for Hep par 1, Glypican-3, Arginase, CA 19-9, CK 19, CDX2, pCEA, SATB2, and Ki-67 having 70% positivity confirming the HCC. Among these IHC panels, all are specific markers of HCC, but CDX2 and SATB2 were aberrantly expressed in our case. He was started on six cycles of chemotherapy (apristar 125 mg, epirubicin 40 mg, oxaliplatin 100 mg, and capecitabine 500 mg). After 8 months of follow-up, he was symptomatically improved. However, later, the patient was lost to follow-up. The accurate pretreatment staging and then providing stage-appropriate treatment is crucial in optimizing esophageal and hepatocellular cancer outcomes. Cases of premortem-diagnosed esophageal metastasis from HCC are extremely rare. Our case was ideal for IHC, which plays an important role in arriving at proper cases. Furthermore, it confirmed and highlighted the rare manifestations of hepatocellular carcinoma.

Thyroid

Histopathological findings of solitary thyroid nodule: An institutional retrospective analysis Harendra Kumar Gautam, Suredrendra Kumar Kanaujia, Vinod Kumar, Devendra Maurya, Shiromani Singh International Journal of Head and Neck Pathology 2018 1(2):37-40 Background: Thyroid swellings are commonly encountered in the surgical practice and accurate preoperative evaluation of thyroid disorder becomes mandatory for the proper management of the patients. Aims and Objectives: The aim of this study was to study the histopathological finding in patients of solitary thyroid nodule at tertiary center of Kanpur region. Materials and Methods: A total of 100 patients with solitary thyroid nodules were thoroughly examined clinically after taking detail history and fine-needle aspiration cytology. All the patients were subjected to surgery after preoperative preparation and anesthesia checkup. Thyroidectomy specimen was evaluated by histopathological examination. Results: Majority of the patients were between the age group of 21 and 40 years. Female:male ratio was about 5.6:1. Swelling in front of the neck was the most common presentation. Most common solitary thyroid swelling was the colloid goiter. The most common surgery performed was hemithyroidectomy. On histopathological examination, a total of 98 out of total 100 patients presented with benign lesions which included 66 patients of colloid goiter, 16 with follicular adenoma, 4 with lymphocytic thyroiditis, and 12 with adenomatous goiter. Two out of total 100 patients had malignant lesions. Among malignant lesions, papillary carcinoma was found in one patient and anaplastic carcinoma in one patient. Conclusion: The most common histopathological findings of solitary thyroid nodule were colloid goiter followed by follicular adenoma, adenomatous goiter and thyroiditis. Thyroid malignancy was very rare histopathological finding.

BRAF status in the variants of papillary thyroid carcinoma Daphne Fonseca, Sudha S Murthy, Ravindranath Tagore, Vishal Rao, Chandrashekar S Rao, K V. V N. Raju, Hemant Kumar Nemade, Sundaram Challa International Journal of Head and Neck Pathology 2018 1(2):41-47 Aim: The aim was to study the BRAF status by immunohistochemistry (IHC) in the variants of papillary carcinoma thyroid and compare it with the clinicopathological parameters. Materials and Methods: All the thyroid carcinomas diagnosed during the period of January 2015–June 2018 were reviewed and classified according to the WHO 2017 criteria. The demographic and clinicopathological features were noted. Microarrays were prepared on 27 cases, including classic and variants of papillary thyroid carcinoma (PTC), poorly differentiated thyroid carcinoma (PDTC), and medullary thyroid carcinoma (MTC). IHC was performed with BRAF V600E by automated staining. The BRAF status was correlated with known prognostic markers. Results: There were 23 PTC, 3 PDTC, and one MTC. The PTC included seven classic, three solid, two each of microcarcinoma, infiltrative and encapsulated follicular variant, tall-cell variant (TCV), oncocytic and one each of diffuse sclerosing, nodular fasciitis-like stroma, and Warthin-like variants. BRAF positivity was seen in 44.44%, including 11 PTC and one PDTC. The positivity was 85.71% in classic and 31.25% in variants. The age (>45 vs. <45 years), gender (male vs. female), number of lesions (unifocal vs. multifocal), type of tumor (PTC vs. other tumors), subtype of PTC (classic PTC vs. variants), invasion (capsular vs. lymphovascular), and aggressive features (extrathyroidal extension vs. lymph nodal involvement) between BRAF positive and negative tumors were not statistically significant (Fisher's exact test at P < 0.05). Conclusion: BRAF status did not show correlation with known prognostic variables in classic as well as variants of PTC.

Head and Neck Pathology

Histopathological findings of solitary thyroid nodule: An institutional retrospective analysis Harendra Kumar Gautam, Suredrendra Kumar Kanaujia, Vinod Kumar, Devendra Maurya, Shiromani Singh International Journal of Head and Neck Pathology 2018 1(2):37-40 Background: Thyroid swellings are commonly encountered in the surgical practice and accurate preoperative evaluation of thyroid disorder becomes mandatory for the proper management of the patients. Aims and Objectives: The aim of this study was to study the histopathological finding in patients of solitary thyroid nodule at tertiary center of Kanpur region. Materials and Methods: A total of 100 patients with solitary thyroid nodules were thoroughly examined clinically after taking detail history and fine-needle aspiration cytology. All the patients were subjected to surgery after preoperative preparation and anesthesia checkup. Thyroidectomy specimen was evaluated by histopathological examination. Results: Majority of the patients were between the age group of 21 and 40 years. Female:male ratio was about 5.6:1. Swelling in front of the neck was the most common presentation. Most common solitary thyroid swelling was the colloid goiter. The most common surgery performed was hemithyroidectomy. On histopathological examination, a total of 98 out of total 100 patients presented with benign lesions which included 66 patients of colloid goiter, 16 with follicular adenoma, 4 with lymphocytic thyroiditis, and 12 with adenomatous goiter. Two out of total 100 patients had malignant lesions. Among malignant lesions, papillary carcinoma was found in one patient and anaplastic carcinoma in one patient. Conclusion: The most common histopathological findings of solitary thyroid nodule were colloid goiter followed by follicular adenoma, adenomatous goiter and thyroiditis. Thyroid malignancy was very rare histopathological finding.

BRAF status in the variants of papillary thyroid carcinoma Daphne Fonseca, Sudha S Murthy, Ravindranath Tagore, Vishal Rao, Chandrashekar S Rao, K V. V N. Raju, Hemant Kumar Nemade, Sundaram Challa International Journal of Head and Neck Pathology 2018 1(2):41-47 Aim: The aim was to study the BRAF status by immunohistochemistry (IHC) in the variants of papillary carcinoma thyroid and compare it with the clinicopathological parameters. Materials and Methods: All the thyroid carcinomas diagnosed during the period of January 2015–June 2018 were reviewed and classified according to the WHO 2017 criteria. The demographic and clinicopathological features were noted. Microarrays were prepared on 27 cases, including classic and variants of papillary thyroid carcinoma (PTC), poorly differentiated thyroid carcinoma (PDTC), and medullary thyroid carcinoma (MTC). IHC was performed with BRAF V600E by automated staining. The BRAF status was correlated with known prognostic markers. Results: There were 23 PTC, 3 PDTC, and one MTC. The PTC included seven classic, three solid, two each of microcarcinoma, infiltrative and encapsulated follicular variant, tall-cell variant (TCV), oncocytic and one each of diffuse sclerosing, nodular fasciitis-like stroma, and Warthin-like variants. BRAF positivity was seen in 44.44%, including 11 PTC and one PDTC. The positivity was 85.71% in classic and 31.25% in variants. The age (>45 vs. <45 years), gender (male vs. female), number of lesions (unifocal vs. multifocal), type of tumor (PTC vs. other tumors), subtype of PTC (classic PTC vs. variants), invasion (capsular vs. lymphovascular), and aggressive features (extrathyroidal extension vs. lymph nodal involvement) between BRAF positive and negative tumors were not statistically significant (Fisher's exact test at P < 0.05). Conclusion: BRAF status did not show correlation with known prognostic variables in classic as well as variants of PTC.

Oral mucosal melanoma: An enigma to the clinician Krishna Sireesha Sundaragiri, Shikha Saxena, Chandni Shekhawat, Bharat Sankhla International Journal of Head and Neck Pathology 2018 1(2):48-51 Oral mucosal melanoma is a rare oral malignancy with well defined clinical presentation of a pigmented brown macular to nodular lesion. The careful histopathological examination along with immunohistochemistry of the biopsy specimen was helpful in identification of this noxious lesion. A clear distinction between management and prognosis of benign gingival lesions and rarer lesions exists. Thus every general dental practitioner should advocate a mandatory histopathological examination of the each and every gingival growth. We present a unique case of oral mucosal melanoma masquerading as a benign gingival growth without any clinical presence of pigmentation in a 58-year-old male patient.

Oral squamous cell carcinoma with myiasis Sakshi Gupta, Arjun Agarwal, Cheena Garg, KY Giri, Archana Chaurasia International Journal of Head and Neck Pathology 2018 1(2):52-54 Myiasis is a rare condition that occurred with advanced ulcerated lesions of the oral cavity caused by the larvae in human tissue that evolve to a parasite and is commonly found in tropical countries. Poor oral hygiene is a primary cause for the growth of maggots leading to the development of myiasis. These cases are reported with chief complaint of pain and irritation in a localized area and diagnosed clinically based on the presence of the maggots. Treatment option is manual removal of maggots to cure the inflammatory process and prevent secondary infections. Here, we present a case of oral squamous cell carcinoma with myiasis diagnosed in a 50-year-old female patient.

A rare case of isolated retro-orbital fungal granuloma Mahmood Dhahir Al-Mendalawi International Journal of Head and Neck Pathology 2018 1(2):55-55

Internal Medicine

Is neutrophil-lymphocyte ratio a novel biomarker for macrovascular and microvascular complications of type 2 diabetes? Walaa H Mohammad, Ahmad B Ahmad, Muhammad H Al-Maghraby, Mohamed Z Abdelrhman, Sanaa Ezzate The Egyptian Journal of Internal Medicine 2019 31(1):1-7 Background/aim The neutrophil-to-lymphocyte (N/L) ratio is a biomarker of inflammation. We aimed to test the hypothesis that macrovascular and microvascular complications of type 2 diabetes mellitus (DM) involve an inflammatory process. Patients and methods This is a cross-sectional study conducted on patients with type 2 DM. Demographic and clinical examination data of eligible patients were recorded. The blood samples for laboratory variables were collected and measured by the standard methods. A standard ultrasound examination was done to measure both carotid artery intima-media thickness (cIMT), and the averages of three results of each side were recorded. Results The current study showed that N/L ratio was an independent predictor of cIMT in patients with type 2 DM (B=0.003, t=8.325, P=0.000), in addition to other conventional cardiovascular risk factors including age, male sex, smoking index, duration of diabetes, estimated glomerular filtration rate, low-density lipoprotein-cholesterol, urinary albumin creatinine ratio, and HBA1c. Moreover, N/L ratio was an independent predictor of albuminuria in patients with type 2 DM (B=0.110, t=3.638, P=0.006), in addition to duration of diabetes, HBA1c, SBP, BMI, and estimated glomerular filtration rate. Conclusion N/L ratio was an independent predictor of cIMT and albuminuria. This supports the hypothesis that diabetic macrovascular and microvascular complications involve an inflammatory process. Therefore, NLR may serve as a cost-effective and readily accessible marker of diabetic vascular complications.

Genetic variants of two-pore calcium channel 2 rs1551305 and its association with type 2 diabetes risk Kareem Essam The Egyptian Journal of Internal Medicine 2019 31(1):8-13 Context The prevalence of diabetes is highest in the Eastern Mediterranean Region, with Egypt leading the region (11% for both sexes) and lowest in the European Region (7% for both sexes). Genome-wide association studies of single-nucleotide polymorphisms (SNPs) have identified a number of variants that are associated with β-cell function and insulin resistance. Two-pore calcium channel 2 (TPCN2) localizes to the lysosome and is a likely receptor for the calcium-mobilizing agent nicotinic acid adenine dinucleotide phosphate. Several studies have indicated that nicotinic acid adenine dinucleotide phosphate may play a role in the insulin signaling of β-cells. Aim of the study The aim of this study was to investigate the association between TPCN2 rs1551305 SNPs and the development of type 2 diabetes. Patients and methods A sample of 158 Egyptian participants was divided into two groups. Group one included 79 type 2 diabetic patients and group two included 79 healthy controls. TPCN2 rs1551305 SNPs were determined by the real-time PCR technique. Results A significant increase in the frequency of G/G genotype in diabetic patients was found. A/A genotype was significantly more frequent in the control group (P=0.001). G allele was also significantly higher in diabetic patients (P=0.008). The G/G genotype showed a 21.37-fold increase in the risk of developing diabetes mellitus. Conclusion The previous findings suggest that TPCN2 rs1551305 SNP is associated with the risk of type 2 diabetes development.

The influence of single-nucleotide polymorphisms of interleukin-1β −511 and +3954 on the susceptibility to Hashimoto's thyroiditis in Egyptian women: immune-endocrine interactions Nearmeen M Rashad, Manar H Soliman, Mayada M Mousa, Azza H Abd El-Fatah The Egyptian Journal of Internal Medicine 2019 31(1):14-21 Background Hashimoto’s thyroiditis (HT) is a T-cell-mediated autoimmune disease. Cytokines play a crucial role in modulating immune responses that affect the balance between maintenance of self-tolerance and initiation of autoimmunity. Thus, we aimed to explore the possible associations of interleukin (IL-1β) −511C/T (rs16944) and IL-1β +3954C/T (rs1143634) gene polymorphisms with susceptibility of HT, and to clarify the impact of these polymorphisms on thyroid function of Egyptian women. Patients and methods Polymorphisms of the IL-1β −511 and IL-1β +3954 genes were assessed in a case–control study comprising 110 HT patients and 90 controls. Genetic variants were genotyped by multiplex PCR. Polymorphisms of the IL-1β were studied by PCR-restriction fragment length polymorphism analysis. Results Our results revealed that the IL-1β −511 CT genotype distribution was significantly higher in HT patients than in controls. With regard to IL-1β +3954 gene polymorphisms, our results showed that there was a nonsignificant difference between the control and HT groups. Women carrying TT and CT genotype of IL-1β −511 had significantly higher values of C-reactive protein, thyroid-stimulating hormone, anti-thyroid peroxidase, and anti-thyroglobulin. As regards the IL-1β serum level, women carrying TT and TC genotype of IL-1β −511 had significantly higher values compared with patients carrying the CC genotype. Moreover, anti-thyroglobulin and free thyroxine were the only independently correlated factors with IL-1β serum level by linear regression analysis. Conclusion CT genotype distribution was significantly higher in patients with HT than in controls with regard to IL-1β −511 (C>T) gene polymorphisms.

The impact of stevioside supplementation on glycemic control and lipid profile in patients with type 2 diabetes: a controlled clinical trial Nearmeen M Rashad, Mona A.E Abdelsamad, Atef M Amer, Mahmoud Z Sitohy, Mayada M Mousa The Egyptian Journal of Internal Medicine 2019 31(1):22-30 Background Stevia is a naturally occurring non-nutritive sweetener that has been reported as sugar substitutes for diabetic patients. We aimed to assess the impact of stevia supplementation on glycemic control in patients with type 2 diabetes mellitus (T2DM). Also, we aimed to examine the association between stevia supplementation and anthropometric measures as well as lipid profile in both obese and diabetic patients. Patients and methods The controlled clinical trial included unrelated 150 participants; 40 patients with T2DM and 60 obese patients and 50 healthy controls. Obese patients were then subdivided into two subgroups according to their fasting blood sugar: nondiabetic (n=30) and 30 patients with T2DM. The participants received stevia (4 mg/kg/body weight) as an alternative to artificial sweetener for 24 weeks. Results Our results found that stevioside supplementation for diabetic patients’ increased the total caloric intake and decreased BMI, waist circumference, waist–hip ratio, and fat mass index, in the obese group. Our results have shown a significant increase of BMI, waist circumference, waist–hip ratio, and fat mass index after 24 weeks of stevia supplementation. In the diabetic group, stevioside for 24 weeks improved the lipid profile and glycemic control, fasting plasma glucose, 2-h plasma glucose, fasting serum insulin, homeostasis model assessment of insulin resistance and hemoglobin A1c (HbA1c), as well as total cholesterol, triglycerides, low density lipoprotein-cholesterol, and high density lipoprotein-cholesterol in all studied intervention groups. Logistic regression test revealed that among clinical and laboratory waist circumference, fasting plasma glucose and HbA1c were independent predictors of response to stevioside. Conclusion Stevioside supplementations for 24 weeks improved cardiometabolic risk in diabetic patients. However, in the obese group, stevioside supplementations increased body weight.

Serum macrophage migration inhibitory factor levels in Hashimoto's thyroiditis MN Ayman Abd Elrahman, Nagwa S Said, Azza Moustafa The Egyptian Journal of Internal Medicine 2019 31(1):31-37 Background Hashimoto’s thyroiditis (HT) is the most common autoimmune thyroid disease. The actual mechanism by which the immunological factors lead to HT has remained obscure. Macrophage migration inhibitory factor (MIF) is recently recognized as a cytokine that has a broad range of inflammatory and immune effects. The aim of this study is to assess the role of MIF in the pathogenesis of HT. Patients and methods One hundred and twenty-nine patients were classified into three groups: (a) overt hypothyroidism, (b) subclinical hypothyroidism, and (c) the control group. A complete history taking, clinical examination, and laboratory investigations were done. Laboratory investigations included thyroid function tests [thyroid-stimulating hormone (TSH), triiodothyronine, and thyroxine], antithyroid antibodies (antithyroid peroxidase antibodies and thyroglobulin antibodies), and MIF level. Results There is highly statistically significant difference between the three groups regarding MIF, the highest level being in group 1>group 2>group 3. In both hypothyroid groups (1 and 2), there are statistically significant positive correlations between MIF with TSH, thyroid peroxidase antibodies, and thyroglobulin antibodies, but there is no statistically significant correlation between MIF with thyroid functions tests and antithyroid antibodies in the control group. Conclusion Serum MIF levels increased in overt and subclinical hypothyroidism and its levels were positively correlated to TSH and thyroid-specific autoantibodies. Serum MIF levels play a role in the pathogenesis of thyroid autoimmune responses in patients with HT. We recommend further larger studies to assess the MIF role in the prognosis of the HT and its treatment by a new strategy.

Salivary versus serum testosterone levels in boys with constitutional delay of growth and puberty Eman H El-Adawy, Enas T El-Khamisy, Ghada El-Said, Amal Seleem The Egyptian Journal of Internal Medicine 2019 31(1):38-42 Background Salivary testosterone (Sal-T) has been established as a noninvasive biomarker in the diagnosis of androgen deficiency in men. However, available data on its utility in adolescent boys with constitutional delay of growth and puberty (CDGP) are not conclusive. Our study was designed to compare salivary versus serum measurements of free testosterone (FT) from the samples collected simultaneously and correlated them with the clinical parameters in boys with CDGP. Patients and methods The study enrolled 25 adolescent boys with CDGP and 20 healthy controls matched for age and sex. Weight, height, BMI, testicular volume, bone age, serum follicle-stimulating hormone (FSH), and luteinizing hormone (LH) were assessed. Simultaneous morning saliva and serum samples were obtained for FT measurements by radioimmunoassay and Sal-T by the enzyme-linked immunosorbent assay technique. Results Adolescent boys with CDGP had significantly lower Sal-T and serum free-T than normal controls (P<0.001). Sal-T positively correlated with BMI, bone age, testicular volume, serum FSH, LH, and FT. With multiple regression analysis BMI, FSH, LH, and serum FT remained independently correlated with Sal-T. Using the receiver operating characteristic curve, Sal-T was found to be more sensitive than serum FT (76 vs. 69%) in the clinical assessment of boys with CDGP. Conclusion Our data shows that Sal-T strongly correlates with FT and provides a sensitive, simple, noninvasive, and diagnostic approach for androgen status in adolescent boys with CDGP.

The relationship between the level of serum-free testosterone and depression in obese adolescent men in Sharkia Governorate MN Ayman Abd-Elrahman, Amira A Fouad, Hossam E Salah The Egyptian Journal of Internal Medicine 2019 31(1):43-51 Background Obesity among children and adolescents is considered as one of the most serious public health concerns; low levels of testosterone have been associated with higher levels of morbidity and may have been associated with depression. Objective To assess the relationship between the level of serum-free testosterone and depression in obese adolescent men in Sharkia Governorate. Patients and methods The present study was conducted on 240 age-matched healthy Egyptian adolescent men, collected from the obesity clinic, Internal Medicine Department, Zagazig University Hospital and five high schools in Sharkia Governorate; all the included participants were subjected to: (a) history taking; (b) physical examination (weight, height, BMI, waist circumference, waist to hip ratio, and Tanner stage using Prader orchidometer); (c) hormonal investigations (free testosterone, estradiol, follicular-stimulating hormone, and luteinizing hormone); (d) psychological assessment for depression using: Mini International Neuropsychiatric Interview for Children and Adolescents and children’s depression inventory. Results There was a significant inverse correlation between free testosterone levels and waist circumference among both control and obese cases (r=−0.31, P=0.031 and r=−0.30, P=0.01, respectively). The free testosterone level was significantly lower in obese cases with depression (2.81±2.32 pg/ml) in comparison with obese cases without depression (3.63±2.65 pg/ml) (P=0.04) and testosterone levels was significantly lower in obese cases having some depressive symptoms (feeling depressed, feeling restless, feeling guilty, eating disorders, sleeping disorders, feeling restless, and suicidality). There were significant inverse correlations (P<0.05) between mean free testosterone level and both mild and moderate depression (r=−0.45, P=0.02 and r=−0.58, P=0.012, respectively), and with the following depressive symptoms; feeling depressed, feeling restless, feeling guilty, eating disorder, sleeping disorders, poor concentration, and suicidality. Conclusion Obese adolescent men in Sharkia Governorate are significantly associated with lower free testosterone levels. Most of the depression symptoms and their degree are inversely correlated with the levels of free testosterone, otherwise the levels of luteinizing hormone, follicular-stimulating hormone, and estradiol showed no correlation with the depression symptoms or their degree. We recommend further larger studies to prove the relationship between the level of serum-free testosterone and depression in obese adolescent men and try to put a management protocol.

Serum β-CrossLaps as a predictor for osteoporosis in postmenopausal women with early diabetic nephropathy Ashraf Mohamed The Egyptian Journal of Internal Medicine 2019 31(1):52-56 Background Osteoporosis is a progressive bone disease with an increased risk of fracture. Objective To estimate bone turnover through an early marker, β-CrossLaps (β-CTx), in postmenopausal women with type 2 diabetes mellitus (DM) and early diabetic nephropathy (DN). Patients and methods A case–control study comprised 80 Egyptian postmenopausal nonsmoker women: 26 women without DM (group 1) as a control group, 28 women had type 2 DM without DN (group 2), and 26 woman were type 2 DM with early DN (group 3). All demographic and clinical data were collected. Laboratory tests including serum calcium, phosphorus, alkaline phosphatase (ALP), bone ALP, β-CTx, glycated hemoglobin, microalbuminuria, serum creatinine, and blood sugar levels were estimated. Results Regarding bone minerals, there was a significant lower serum level of calcium and phosphorus in group 3 compared with groups 1 and 2 (P=0.029 and <0.0001, respectively). Groups 2 and 3 had significantly lower serum levels of total ALP (P=0.011 and 0.006, respectively) and bone ALP (with lowest level in group 3) (P<0.0001) compared with group 1. Groups 2 and 3 had significantly higher serum levels of β-CTx (with highest level in group 3) compared with group 1 (P=0.016 and 0.003, respectively). Dual-energy X-ray absorptiometry scan revealed a significant elevation in osteoporosis percent in group 3 (63.4%) in comparison with group 1 (26.9%) (P=0.001), whereas osteopenia percent was significantly higher in group 2 (P=0.004) compared with group 3. Conclusion In diabetic postmenopausal women, serum β-CTx and microalbuminuria are potentially useful tools for assessing bone resorption state.

Association of serum growth differentiation factor 15 and hepatocellular carcinoma in Egyptian patients Mohamed-Naguib Wifi, Rania A Zayed, Nevine Fouad, Ahmed Y Hassan, Maha A Hussien, Mohamed G Sokar The Egyptian Journal of Internal Medicine 2019 31(1):57-63 Background Liver cirrhosis and hepatocellular carcinoma (HCC) are consequences of chronic hepatitis C virus (HCV) infection. HCC is one of the fastest rising causes of cancer-related mortality. This dismal prognosis is related to late diagnosis with currently available screening methods. The aim of our study was to evaluate the diagnostic power of serum growth differentiation factor (GDF) 15 in HCC detection and its ability to distinct HCC from cirrhosis in chronic hepatitis C Egyptian patients. Patients and methods Ninety participants were included in the study; 30 patients with HCV-cirrhosis, 30 patients with HCV-Cirrhosis and HCC, and 30 gender and age-matched healthy subjects as the control group. The patients were subjected to history taking, clinical examinations, routine laboratory analysis, and α-fetoprotein (AFP) determination. Serum GDF15 was measured using an enzyme-linked immunosorbent assay kit. Results The mean level of GDF15 in HCV-cirrhosis patients was 140.28±128.66 pg/ml, in HCV-HCC patients 154.45±123.74 pg/ml, and in the control group it was 81.19±42.53 pg/ml. Statistically significant difference in GDF15 level was found between HCV-HCC patients and controls, P=0.012, while no statistically significant difference was found on comparing HCV-cirrhosis patients to controls or to HCV-HCC patients, P=0.064 and 0.473, respectively. The cut-off value of GDF15 to discriminate HCV-HCC patients from controls was 122.3 pg/ml with 53.3% sensitivity and 86.7% specificity and an area under the receiver operating characteristic (AUROC) of 0.692. AFP at a cut-off value of 20.85 ng/l was able to discriminate HCV-HCC patients from HCV-cirrhosis patients with 73.3% sensitivity and 73.3% specificity and an AUROC of 0.744. AUROC for combined AFP and GDF15 showed lower performance than AFP alone in discrimination of HCV-HCC from HCV-cirrhosis patients (AUROC=0.642). Conclusion GDF15 is not a potential diagnostic marker for the distinction of HCC from cirrhosis in chronic hepatitis C Egyptian patients.

The role of hepatitis C virus and possible risk factors in development of hepatocellular carcinoma: 400 patients based study Fatema Abu-bakr Abdelmoez, Hala Mostafa Imam, Naglaa kamal Idriss, Lobna Abdel Wahid, Wael Ahmed Abbas, Mohamed Abozaid Ali Abozaid, Hossam Mahmoud Abdelwahab The Egyptian Journal of Internal Medicine 2019 31(1):64-72 Background and aims Hepatocellular carcinoma (HCC) is one of the commonest tumors and considered the fifth most common malignant neoplasm and a major cause of death. Egypt has increased incidence of HCC cases, as Egypt has the highest prevalence of hepatitis C virus (HCV) infection. The aim was to study the epidemiological characteristics of HCC in Assiut, Egypt. Patients and methods A descriptive observational study design was applied for the present study. The studied population was 400 patients with HCC (288 of them were male and 112 were female) who fulfilled the diagnostic criteria for HCC. Data were analyzed for exploring the clinical, etiological, radiological, and tumor characteristics of the studied patients. Results The mean age of the patients was 59.85±9.1 years. Most cases (68%) were from rural areas, 38% of the patients were in agricultural occupation, and 32% of the patients were accidentally discovered to have HCC .The most frequent symptom was abdominal pain (15%). HCV antibody was present in 63% of the patients, HBV infection was recorded in 28%, coinfection was seen in 3%, and no viral infection was present in 6%. Diabetes mellitus was present in 37% and obesity in 24% of the patients. Right lobe of the liver was the most frequent affected lobe (61%), and 69% of the cases had a single lesion. Conclusion HCC incidence had been increasing in the past years in Egypt. The high prevalence of HCV infection in Egypt makes the surveillance strategies important for early detection of HCC in these patients to provide better curative treatment modalities in the early stages.

Angiogenesis

Interleukin-8 release by endothelial colony-forming cells isolated from idiopathic pulmonary fibrosis patients might contribute to their pathogenicity Abstract Introduction Idiopathic pulmonary fibrosis (IPF) is a devastating disease characterized by obliteration of alveolar architecture, resulting in declining lung function and ultimately death. Pathogenic mechanisms involve a concomitant accumulation of scar tissue together with myofibroblasts activation and a strong abnormal vascular remodeling. Endothelial progenitor cells (ECFC subtype) have been investigated in several human lung diseases as a potential actor in IPF. We previously demonstrated that ECFCs are down-regulated in IPF in contrast to healthy controls. We postulated here that ECFCs might behave as a liquid biopsy in IPF patients and that they exert modified vasculogenic properties. Methods and results ECFCs isolated from controls and IPF patients expressed markers of the endothelial lineage and did not differ concerning adhesion, migration, and differentiation in vitro and in vivo. However, senescent and apoptotic states were increased in ECFCs from IPF patients as shown by galactosidase staining, p16 expression, and annexin-V staining. Furthermore, conditioned medium of IPF-ECFCs had increased level of interleukin-8 that induced migration of neutrophils in vitro and in vivo. In addition, an infiltration by neutrophils was shown in IPF lung biopsies and we found in a prospective clinical study that a high level of neutrophils in peripheral blood of IPF patients was associated to a poor prognosis. Conclusion To conclude, our study shows that IPF patients have a senescent ECFC phenotype associated with an increased IL-8 secretion potential that might contribute to lung neutrophils invasion during IPF.

ADAM10 controls the differentiation of the coronary arterial endothelium Abstract The coronary vasculature is crucial for normal heart function, yet much remains to be learned about its development, especially the maturation of coronary arterial endothelium. Here, we show that endothelial inactivation of ADAM10, a key regulator of Notch signaling, leads to defects in coronary arterial differentiation, as evidenced by dysregulated genes related to Notch signaling and arterial identity. Moreover, transcriptome analysis indicated reduced EGFR signaling in A10ΔEC coronary endothelium. Further analysis revealed that A10ΔEC mice have enlarged dysfunctional hearts with abnormal myocardial compaction, and increased expression of venous and immature endothelium markers. These findings provide the first evidence for a potential role for endothelial ADAM10 in cardioprotective homeostatic EGFR signaling and implicate ADAM10/Notch signaling in coronary arterial cell specification, which is vital for normal heart development and function. The ADAM10/Notch signaling pathway thus emerges as a potential therapeutic target for improving the regenerative capacity and maturation of the coronary vasculature.

Interleukin-22 promotes tumor angiogenesis Abstract TH17 cells play important yet complex roles in cancer development and progression. We previously reported that TH17 cells and IL-17 mediate resistance to anti-VEGF therapy by inducing recruitment of immunosuppressive and proangiogenic myeloid cells to the tumor microenvironment. Here, we demonstrate that IL-22, a key effector cytokine expressed by TH17 cells, directly acts on endothelial cells to promote tumor angiogenesis. IL-22 induces endothelial cell proliferation, survival, and chemotaxis in vitro and neovascularization in an ex vivo mouse choroid explant model. Blockade of IL-22, with a neutralizing antibody, significantly inhibits tumor growth associated with reduced microvascular density. No synergistic effect of IL-22 with VEGF was observed. These results identify IL-22 as a potential therapeutic target for blocking tumor angiogenesis.

TSPYL5-mediated inhibition of p53 promotes human endothelial cell function Abstract Testis-specific protein, Y-encoded like (TSPYL) family proteins (TSPYL1-6), which are members of the nucleosome assembly protein superfamily, have been determined to be involved in the regulation of various cellular functions. However, the potential role of TSPYL family proteins in endothelial cells (ECs) has not been determined. Here, we demonstrated that the expression of TSPYL5 is highly enriched in human ECs such as human umbilical vein endothelial cells (HUVECs) and human pluripotent stem cell-differentiated ECs (hPSC-ECs). Importantly, TSPYL5 overexpression was shown to promote EC proliferation and functions, such as migration and tube formation, by downregulating p53 expression. Adriamycin-induced senescence was markedly blocked by TSPYL5 overexpression. In addition, the TSPYL5 depletion-mediated loss of EC functions was blocked by p53 inhibition. Significantly, TSPYL5 overexpression promoted angiogenesis in Matrigel plug and wound repair in a mouse skin wound healing model in vivo. Our results suggest that TSPYL5, a novel angiogenic regulator, plays a key role in maintaining endothelial integrity and function. These findings extend the understanding of TSPYL5-dependent mechanisms underlying the regulation of p53-related functions in ECs.

The regulatory network of miR-141 in the inhibition of angiogenesis Abstract The miR-200 family, consisting of miR-200a/b/c, miR-141, and miR-429, is well known to inhibit epithelial-to-mesenchymal transition (EMT) in cancer invasion and metastasis. Among the miR-200 family members, miR-200a/b/c and miR-429 have been reported to inhibit angiogenesis. However, the role of miR-141 in angiogenesis remains elusive, as contradicting results have been found in different cancer types and tumor models. Particularly, the effect of miR-141 in vascular endothelial cells has not been defined. In this study, we used several in vitro and in vivo models to demonstrate that miR-141 in endothelial cells inhibits angiogenesis. Additional mechanistic studies showed that miR-141 suppresses angiogenesis through multiple targets, including NRP1, GAB1, CXCL12β, TGFβ2, and GATA6, and bioinformatics analysis indicated that miR-141 and its targets comprise a powerful and precise regulatory network to modulate angiogenesis. Taken together, these data not only demonstrate an anti-angiogenic effect of miR-141, further strengthening the critical role of miR-200 family in the process of angiogenesis, but also provides a valuable cancer therapeutic target to control both angiogenesis and EMT, two essential steps in tumor growth and metastasis.

A ribosomal DNA-hosted microRNA regulates zebrafish embryonic angiogenesis Abstract MicroRNAs (miRNAs) are single-stranded small non-coding RNAs, generally 18–25 nucleotides in length, that act as repressors of gene expression. miRNAs are encoded by independent genes or processed from a variety of different RNA species. So far, there is no evidence showing that the ribosomal DNA-hosted microRNA is implicated in vertebrate development. Currently, we found a highly expressed small RNA hosted in ribosomal DNA was predicted as a novel miRNA, named miR-ntu1, in zebrafish endothelial cells by deep sequencing analysis. The miRNA was validated by custom-designed Taqman PCR, Northern Blot, and in silico analysis. Furthermore, we demonstrated that miR-ntu1 played a crucial role in zebrafish angiogenesis via modulation of Notch signaling. Our findings provide a notable case that a miRNA hosted in ribosomal DNA is involved in vertebrate development.

Live imaging of angiogenesis during cutaneous wound healing in adult zebrafish Abstract Angiogenesis, the growth of new blood vessels from pre-existing vessels, is critical for cutaneous wound healing. However, it remains elusive how endothelial cells (ECs) and pericytes (PCs) establish new blood vessels during cutaneous angiogenesis. We set up a live-imaging system to analyze cutaneous angiogenesis in adult zebrafish. First, we characterized basic structures of cutaneous vasculature. In normal skin tissues, ECs and PCs remained dormant to maintain quiescent blood vessels, whereas cutaneous injury immediately induced angiogenesis through the vascular endothelial growth factor signaling pathway. Tortuous and disorganized vessel networks formed within a few weeks after the injury and subsequently normalized through vessel regression in a few months. Analyses of the repair process of injured single blood vessels revealed that severed vessels elongated upon injury and anastomosed with each other. Thereafter, repaired vessels and adjacent uninjured vessels became tortuous by increasing the number of ECs. In parallel, PCs divided and migrated to cover the tortuous blood vessels. ECs sprouted from the PC-covered tortuous vessels, suggesting that EC sprouting does not require PC detachment from the vessel wall. Thus, live imaging of cutaneous angiogenesis in adult zebrafish enables us to clarify how ECs and PCs develop new blood vessels during cutaneous angiogenesis.

AutoTube: a novel software for the automated morphometric analysis of vascular networks in tissues Abstract Due to their involvement in many physiologic and pathologic processes, there is a great interest in identifying new molecular pathways that mediate the formation and function of blood and lymphatic vessels. Vascular research increasingly involves the image-based analysis and quantification of vessel networks in tissue whole-mounts or of tube-like structures formed by cultured endothelial cells in vitro. While both types of experiments deliver important mechanistic insights into (lymph)angiogenic processes, the manual analysis and quantification of such experiments are typically labour-intensive and affected by inter-experimenter variability. To bypass these problems, we developed AutoTube, a new software that quantifies parameters like the area covered by vessels, vessel width, skeleton length and branching or crossing points of vascular networks in tissues and in in vitro assays. AutoTube is freely downloadable, comprises an intuitive graphical user interface and helps to perform otherwise highly time-consuming image analyses in a rapid, automated and reproducible manner. By analysing lymphatic and blood vascular networks in whole-mounts prepared from different tissues or from gene-targeted mice with known vascular abnormalities, we demonstrate the ability of AutoTube to determine vascular parameters in close agreement to the manual analyses and to identify statistically significant differences in vascular morphology in tissues and in vascular networks formed in in vitro assays.

Kruppel-like factor 4 regulates developmental angiogenesis through disruption of the RBP-J–NICD–MAML complex in intron 3 of Dll4 Abstract Angiogenesis is a multistep process that requires highly regulated endothelial cell (EC) behavior. The transcription factor Krüppel-like factor 4 (KLF4) is a critical regulator of several basic EC functions; we have recently shown that KLF4 disturbs pathological (tumor) angiogenesis by mediating the expression of members of VEGF and Notch signaling pathways. Notch signaling is central to orchestration of sprouting angiogenesis but little is known about the upstream regulation of Notch itself. To determine the role of KLF4 in normal (developmental) angiogenesis, we used a mouse retinal angiogenesis model. We found that endothelial-specific overexpression of KLF4 in transgenic mice (EC-K4 Tg) leads to increased vessel density, branching and number of tip cell filopodia as assessed on postnatal day 6 (P6). The hypertrophic vasculature seen with sustained KLF4 overexpression is not stable and undergoes prominent remodeling during P7–P12 resulting in a normal appearing retinal vasculature in adult EC-K4 Tg mice. We find that KLF4 inhibits Delta-like 4 (DLL4) expression in the angiogenic front during retinal vascular development. Furthermore, in an oxygen-induced retinopathy model, overexpression of KLF4 results in decreased vaso-obliteration and neovascular tuft formation that is similar to genetic or pharmacologic DLL4 inhibition. Mechanistically, we show that KLF4 disables the activity of the essential Notch transcriptional activator RBP-J by interfering with binding of co-activators NICD and MAML at intron 3 of the Notch ligand DLL4. In summary, our experimental results demonstrate a regulatory role of KLF4 in developmental angiogenesis through regulation of DLL4 transcription.

Mouse models of Alzheimer's disease cause rarefaction of pial collaterals and increased severity of ischemic stroke Abstract Vascular dysfunction contributes to the progression and severity of Alzheimer's disease (AD). Patients with AD also sustain larger infarctions after ischemic stroke; however, the responsible mechanisms are unknown. Pial collaterals are the primary source of protection in stroke. Unfortunately, natural aging and other vascular risk factors cause a decline in collateral number and diameter (rarefaction) and an increase in stroke severity. Herein, we tested the hypothesis that AD accelerates age-induced collateral rarefaction and examined potential underlying mechanisms. Triple and double transgenic mouse models of AD both sustained collateral rarefaction by 8 months of age, well before the onset of rarefaction caused by aging alone (16 months of age). Rarefaction, which did not progress further at 18 months of age, was accompanied by a twofold increase in infarct volume after MCA occlusion. AD did not induce rarefaction of similarly sized pial arterioles or penetrating arterioles. Rarefaction was minimal and occurred only at 18 months of age in a parenchymal vascular amyloid-beta model of AD. Rarefaction was not associated with amyloid-beta deposition on collaterals or pial arteries, nor was plaque burden or CD11b+ cell density greater in brain underlying the collateral zones versus elsewhere. However, rarefaction was accompanied by increased markers of oxidative stress, inflammation, and aging of collateral endothelial and mural cells. Moreover, rarefaction was lessened by deletion of CX3CR1 and prevented by overexpression of eNOS. These findings demonstrate that mouse models of AD promote rarefaction of pial collaterals and implicate inflammation-induced accelerated aging of collateral wall cells. Strategies that reduce vascular inflammation and/or increase nitric oxide may preserve collateral function.