Clinical Immunology

iPLA 2 Activation Mediates Granular Exocytosis and Corrects Microbicidal Defects in ROS-Deficient and CGD Human Neutrophils Abstract Purpose The ubiquitous calcium–independent phospholipase A2 enzyme (iPLA2) is inhibited by calmodulin binding and known to be responsible for phospholipid remodeling housekeeping functions including granule exocytosis–associated membrane fusion in normal human neutrophils. We evaluate in human neutrophils the iPLA2 secretagogue effects using normal neutrophils, where reactive oxygen species (ROS) generation has been blocked by diphenyleneiodonium, as well as in neutrophils from chronic granulomatous disease (CGD) patients. Methods Neutrophils were pretreated with W7, a calmodulin inhibitor known to activate iPLA2 and exocytosis of granules, and vesicles as well as intra- and extra-microbicidal activity against Staphylococcus aureus and Aspergillus fumigatus were evaluated. Results W7 increases exocytosis of primary, secondary, and tertiary granules and vesicles and improves neutrophil microbicidal activity against S. aureus and A. fumigatus. Conclusions In neutrophils, calmodulin-mediated iPLA2 inhibition controls granule and vesicle exocytosis in the phagosome and in the extracellular microenvironment. Relieving iPLA2 inhibition results in increased exocytosis of primary, secondary, and tertiary granules and secretory vesicles with correction of defective intracellular and extracellular microbicidal activity. In CGD patients presenting ROS defective production, this increase in the non-oxidative killing pathway partially corrects their microbicidal defects.

Different Phenotypic Presentations of X-Linked Lymphoproliferative Disease in Siblings with Identical Mutations

Efficacy and Adverse Events During Janus Kinase Inhibitor Treatment of SAVI Syndrome Abstract Objectives Mutations affecting the TMEM173 gene cause STING-associated vasculopathy with onset in infancy (SAVI). No standard immunosuppressive treatment approach is able to control disease progression in patients with SAVI. We studied the efficacy and safety of targeting type I IFN signaling with the Janus kinase inhibitor, ruxolitinib. Methods We used DNA sequencing to identify mutations in TMEM173 in patients with peripheral blood type I IFN signature. The JAK1/2 inhibitor ruxolitinib was administered on an off-label basis. Results We identified three patients with SAVI presenting with skin involvement and progressive severe interstitial lung disease. Indirect echocardiographic signs of pulmonary hypertension were present in one case. Following treatment with ruxolitinib, we observed improvements of respiratory function including increased forced vital capacity in two patients, with discontinuation of oxygen therapy and resolution of echocardiographic abnormalities in one case. Efficacy was persistent in one patient and only transitory in the other two patients. Clinical control of skin complications was obtained, and one patient discontinued steroid treatment. One patient, who presented with kidney involvement, showed resolution of hematuria. One patient experienced increased recurrence of severe viral respiratory infections. Monitoring of peripheral blood type I interferon signature during ruxolitinib treatment did not show a stable decrease. Conclusions We conclude that targeting type I IFN receptor signaling may represent a promising therapeutic option for a subset of patients with SAVI syndrome and severe lung involvement. However, the occurrence of viral respiratory infection might represent an important cautionary note for the application of such form of treatment.

Clinical and Laboratory Features of 184 Italian Pediatric Patients Affected with Selective IgA Deficiency (SIgAD): a Longitudinal Single-Center Study Abstract Purpose Selective IgA deficiency (SIgAD) is the most common humoral primary immunodeficiency. Long-term follow-up data in large cohort of pediatric patients are scarce. Methods We report on a single-center cohort of 184 pediatric patients affected with selective IgA deficiency and describe the characteristics at diagnosis and during follow-up. Results Respiratory infections were the most common clinical finding leading to the initial diagnosis (62%). Positive family history for antibody deficiencies (selective IgA deficiency, common variable immunodeficiency) led to SIgAD diagnosis in 16% of cases. During follow-up, while the incidence of respiratory infections was not particularly high, gastrointestinal symptoms were reported in 27% of patients. Allergic manifestations were found in 23% at diagnosis and an additional 16% of patients during follow-up, leading to a prevalence of atopy of 39% among SIgAD patients. Autoimmune manifestations, excluding celiac disease, were found in 9% of affected patients during follow-up. Celiac disease was found in a high prevalence (14%). Increase of serum IgA levels to partial deficiency (9%) and normal serum levels for age (4%) was observed during follow-up. A small percentage of patients (2%) progressed to common variable immunodeficiency (CVID). Conclusions In conclusion, this is the first study to describe a large single-center pediatric cohort of patients affected with SIgAD, revealing that overall most patients do well with regard to infections. Many develop CD, at a rate much higher than the general population. A few normalize their IgA levels. A few progress to CVID. Thus, careful follow-up is suggested to diagnose and treat potential complications earlier for avoiding potential morbidities.

Autoinflammation Masquerading as Autoimmunity in an Adult with Heterozygous p.E250K PSTPIP1 Mutation

Successful Lung Transplantation in a Patient with Chronic Granulomatous Disease

Enterovirus-Associated HLH: Addition of Anakinra to IVIG and Corticosteroids

Defining Primary Selective IgM Deficiency

Variable Responses to Tocilizumab in Four Patients with Schnitzler Syndrome

Role of Allogeneic Hematopoietic Stem Cell Transplant for Chronic Granulomatous Disease (CGD): a Report of the United States Immunodeficiency Network Abstract Purpose Chronic granulomatous disease (CGD) is a primary immunodeficiency for which allogeneic hematopoietic stem cell transplant (HSCT) offers potential cure. Direct comparison of HSCT to non-HSCT management in the North American population was performed to identify clinical factors associated with overall survival (OS) and transplant-related survival (TRS). Methods Retrospective review of CGD subjects enrolled in the United States Immunodeficiency Network. Survival was estimated by the Kaplan-Meier method and modeled by proportional hazards regression. Results We identified 507 patients (66% CYBB mutants) diagnosed in 1953–2016. Fifty underwent allogeneic HSCT. Median follow-up was 9.1 years after diagnosis (0–45.8 years). OS was negatively associated with CYBBmutation (HR = 6.25; p = 0.034) and not associated with HSCT (88% v. 85% ± HCT) (HR = 1.26; p = 0.65). Transplant at ≤ 14 years old was associated with improved TRS (93% v. 82% at T + 60 months) (HR = − 4.51; p = 0.035). Patients transplanted before 15 years old had fewer severe infections pre-HSCT (mean 0.95 v. 2.13; p = 0.047). No mortality was reported in patients receiving stem cells from matched siblings. Infection incidence declined post-HSCT in subjects with greater than or equal to four infections pre-HSCT (p = 0.0010). Compared to non-HSCT patients ≥ 15 years old, post-transplant survivors had higher mean performance score (93.2 v. 85.9; p = 0.0039) and lower frequency of disability (11% v. 52%; p = 0.014). Conclusion Allogeneic HSCT was associated with reduced infection incidence and improved functional performance, but not with a change in overall survival. Transplant-related survival was elevated in patients undergoing HSCT before 15 years old. Consider HSCT prior to late adolescence in patients with severely diminished reactive oxygen intermediate synthesis, particularly if a matched sibling is available.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Nuclear Medicine

Preliminary results of biodistribution and dosimetric analysis of [ 68 Ga]Ga-DOTA ZOL : a new zoledronate-based bisphosphonate for PET/CT diagnosis of bone diseases Abstract Objective Pre-clinical studies with gallium-68 zoledronate ([68Ga]Ga-DOTAZOL) have proposed it to be a potent bisphosphonate for PET/CT diagnosis of bone diseases and diagnostic counterpart to [177Lu]Lu-DOTAZOL and [225Ac]Ac-DOTAZOL. This study aims to be the first human biodistribution and dosimetric analysis of [68Ga]Ga-DOTAZOL. Methods Five metastatic skeletal disease patients (mean age: 72 years, M: F; 4:1) were injected with 150–190 MBq (4.05–5.14 mCi) of [68Ga]Ga-DOTAZOL i.v. Biodistribution of [68Ga]Ga-DOTAZOL was studied with PET/CT initial dynamic imaging for 30 min; list mode over abdomen (reconstructed as six images of 300 s) followed by static (skull to mid-thigh) imaging at 45 min and 2.5 h with Siemens Biograph 2 PET/CT camera. Also, blood samples (8 time points) and urine samples (2 time points) were collected over a period of 2.5 h. Total activity (MBq) in source organs was determined using interview fusion software (MEDISO Medical Imaging Systems, Budapest, Hungary). A blood-based method for bone marrow self-dose determination and a trapezoidal method for urinary bladder contents residence time calculation were used. OLINDA/EXM version 2.0 software (Hermes Medical Solutions, Stockholm, Sweden) was used to generate residence times for source organs, organ absorbed doses and effective doses. Results High uptake in skeleton as target organ, kidneys and urinary bladder as organs of excretion and faint uptake in liver, spleen and salivary glands were seen. Qualitative and quantitative analysis supported fast blood clearance, high bone to soft tissue and lesion to normal bone uptake with [68Ga]Ga-DOTAZOL. Urinary bladder with the highest absorbed dose of 0.368 mSv/MBq presented the critical organ, followed by osteogenic cells, kidneys and red marrow receiving doses of 0.040, 0.031 and 0.027 mSv/MBq, respectively. The mean effective dose was found to be 0.0174 mSv/MBq which results in an effective dose of 2.61 mSv from 150 MBq. Conclusions Biodistribution of [68Ga]Ga-DOTAZOL was comparable to [18F]NaF, [99mTc]Tc-MDP and [68Ga]Ga-PSMA-617. With proper hydration and diuresis to reduce urinary bladder and kidney absorbed doses, it has clear advantages over [18F]NaF owing to its onsite, low-cost production and theranostic potential of personalized dosimetry for treatment with [177Lu]Lu-DOTAZOL and [225Ac]Ac-DOTAZOL.

Preliminary clinical assessment of dynamic 18 F-fluorodeoxyglucose positron emission tomography/computed tomography for evaluating lymph node metastasis in patients with lung cancer: a prospective study Abstract Objective We assessed the diagnostic capacity of dynamic fluorine-18-fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) and dual-time-point (DTP) PET/CT to explore the optimal scan timing for nodal staging in lung cancer. Methods Thirty-four patients with lung cancer underwent dynamic and consecutive DTP PET/CT scans. Two readers visually evaluated FDG uptake within each lymph node (LN) and pulmonary artery (metastatic LN: n = 10; nonmetastatic LN: n = 121). For each dynamic and DTP scan, we compared the maximum standardized uptake value (SUVmax) and the retention index of the SUVmax (RI-SUVmax) between metastatic and nonmetastatic LNs. We compared the diagnostic capacity of the dynamic and DTP scans using receiver operating characteristic (ROC) analyses. Results In the visual analyses of LN metastases, a sensitivity of 20.0–60.0% and specificity of 97.5–100.0% were identified for the first to third dynamic scans. The sensitivity of the 1-h early and 2-h delayed scans was 80.0% and 90.0%, respectively, whereas the specificity was 66.9% and 47.9%, respectively. The visual analysis of the dynamic second phase had the highest accuracy. Semiquantitative analyses revealed that the SUVmax was significantly higher for metastatic LNs than for nonmetastatic LNs in the dynamic second and third phases and the 1-h early and 2-h delayed phases (p < 0.05 for all). The RI-SUVmax was higher in metastatic LNs than in nonmetastatic LNs for the dynamic scan (p = 0.004) and the DTP scan (p = 0.002). The ROC analyses showed that SUV2 and SUV3 had higher performances with high specificity, high negative predictive value, and high accuracy than the other parameters. The area under the ROC curve of the RI-SUV-dual-time-point had the highest value (0.794) without any significant differences between the area under the ROC curves for all parameters (p > 0.05 for all). Conclusions Based on the visual and semiquantitative analyses, 18F-FDG dynamic PET/CT exhibited excellent performance with extremely high specificity in the dynamic second phase.

Distribution of LAT1-targeting PET tracer was independent of the tumor blood flow in rat xenograft models of C6 glioma and MIA PaCa-2 Abstract Objective L-type amino acid transporter 1 (LAT1) is strongly expressed on the cell membrane in various types of human cancer cells, while being minimally expressed in normal or inflammatory tissues. Therefore, LAT1-targeting PET tracers have been developed for cancer-specific imaging. The purpose of this study was to study the distribution of two LAT1-targeting PET tracers, L-4-borono-2-18F-fluoro-phenylalanine (18F-FBPA) and L-3-18F-alpha-methyl tyrosine (18F-FAMT), in relation to the tumor blood flow, using rat xenograft models. Methods Rat tumor xenograft models of C6 glioma (n = 4; tumors = 8) and MIA PaCa-2 (pancreatic cancer) (n = 4; tumors = 6) were used. The expressions of LAT1 and CD98hc were evaluated by both immunofluorescence staining and western blot analysis. Dynamic PET was performed after injection of 18F-FAMT or 18F-FBPA (scan duration = 70 min) following 15O-water PET (scan duration = 10 min). The PET data were subjected to kinetic analyses, and the K1, k2, and total distribution volume (Vt) were calculated using the one-tissue compartment model. The accumulation of the LAT1 tracers was expressed in terms of their Vt. Tumor blood flow (TBF) was represented by the K1 value in 15O-water PET. Results LAT1/CD98hc expression was confirmed in both xenografts by immunofluorescence staining. Western blot analysis showed higher functional expression of LAT1 in the C6 glioma cells as compared to the MIA PaCa-2 cells (C6 glioma/MIA PaCa-2 relative expression ratio = 1.70). The Vt values of both 18F-FBPA and 18F-FAMT were significantly higher in the C6 glioma xenografts than in the MIA PaCa-2 xenografts (C6 glioma: 2.27 ± 0.35 and 2.03 ± 0.23, respectively; MIA PaCa-2: 1.28 ± 0.26 and 1.35 ± 0.15, respectively). Meanwhile, there was no significant correlation of the Vt value of either 18F-FBPA or 18F-FAMT with the TBF, in either the C6 glioma or the MIA PaCa-2 xenografts. Conclusions This study revealed that total distribution volumes of the LAT1-targeting PET tracers 18F-FBPA and 18F-FAMT were independent of the tumor blood flow and might reflect the functional expression levels of LAT1 in the C6 glioma and MIA PaCa-2 xenograft models.

Characterization of the binding of tau imaging ligands to melanin-containing cells: putative off-target-binding site Abstract Objective Amyloid-β plaques and neurofibrillary tangles composed of tau protein are the neuropathological hallmarks of Alzheimer's disease. In recent years, marked progress has been made in Alzheimer's disease research using tau ligands for positron emission tomography (PET). However, the issue of off-target binding, that is, the binding of ligands to regions without tau pathology, remains unresolved. Tissues with melanin-containing cells (MCCs) have been suggested as binding targets for tau ligands. In the present study, we characterized the MCC-binding properties of representative tau PET ligands. Methods Autoradiographic studies of [18F]AV-1451 and [18F]THK5351 were conducted using postmortem human midbrain sections. Saturation-binding assays of [18F]AV-1451 and [18F]THK5351 were performed with B16F10 melanoma cells. The blocking effects of 25 compounds against [18F]THK5351 binding to B16F10 cells were used to investigate the relationship between chemical structure and MCC binding. Results Autoradiography demonstrated specific binding of the radioligands in the substantia nigra. [18F]AV-1451 and [18F]THK5351 exhibited saturable binding to melanoma cells ([18F]AV-1451: Kd = 669 ± 196 nM, Bmax = 622 ± 269 pmol/mg protein; [18F]THK5351: Kd = 441 ± 126 nM, Bmax = 559 ± 75.5 pmol/mg protein). In blocking studies with melanoma cells, compounds bearing multiple aromatic rings and an aminopyridine group, including tau ligands such as AV-1451, PBB3, and a lead compound of MK-6240, exhibited the inhibition of [18F]THK5351 binding comparable to self-blocking by THK5351 (> 70% at 10 µM). Conclusions These studies suggest that the binding properties of [18F]AV-1451 and [18F]THK5351 are sufficient to expect highlighting of tissues with a high density of MCCs. The findings of the present study should aid the development of neuroimaging ligands that do not bind to MCC.

Potential utility of bone scan in cranial bone flap osteomyelitis Abstract Objective Currently, the diagnosis of bone flap osteomyelitis (BFO) remains a challenge for medical imaging. The present study aimed to identify predictive scintigraphic patterns of BFO. Methods This retrospective study reviewed planar bone scan of patients with suspected BFO between 2010, and 2016. A total of 15 patients were included. Final diagnosis of BFO was obtained by histological and bacteriological documentation. Eight scintigraphic signs potentially helpful were reviewed and correlated with the final diagnosis individually or in combination through Fischer exact test. Results Eight patients out of 15 (53.3%) were diagnosed with BFO. Radionuclide uptake inside the bone flap during blood-pool phase was predictive for BFO (p = 0.007) with 75.0% sensitivity 100% specificity, and 86.7% accuracy. In combination, radionuclide uptake inside the bone flap or a spreading wavefront between blood-pool and delayed phases was associated with BFO (p = 0.007). It did not improve diagnostic performance. Conclusion Using well-defined and reproducible scintigraphic signs, bone scan is helpful for the diagnosis of BFO.

Novel 3D heart left ventricle muscle segmentation method for PET-gated protocol and its verification Abstract Objective The aim of this study was to propose and verify a universal method of left ventricular myocardium segmentation, able to operate on heart gated PET data with different sizes, shapes and uptake distributions. The proposed method can be classified as active model method and is based on the BEAS (B-spline Explicit Active Surface) algorithm published by Barbosa et al. The method was implemented within the Pmod PCARD software package. Method verification by comparison with reference software and phantom data is also presented in the paper. Methods The proposed method extends the BEAS model by defining mechanical features of the model: tensile strength and bending resistance. Formulas describing model internal energy increase during its stretching and bending are proposed. The segmentation model was applied to the data of 60 patients, who had undergone cardiac gated PET scanning. QGS by Cedars-Sinai and ECTb by Emory University Medical Centre served as reference software for comparing ventricular volumes. The method was also verified using data of left ventricular phantoms of known volume. Results The results of the proposed method are well correlated with the results of QGS (slope: 0.841, intercept: 0.944 ml, R2: 0.867) and ECTb (slope: 0.830, intercept: 2.109 ml, R2: 0.845). The volumes calculated by the proposed method were very close to the true cavity volumes of two different phantoms. Conclusions The analysis of gated PET data by the proposed method results in volume measurements comparable to established methods. Phantom experiments demonstrate that the volume values correspond to the physical ones.

The use of 3′-deoxy-3′- 18 F-fluorothymidine (FLT) PET in the assessment of long-term survival in breast cancer patients treated with neoadjuvant chemotherapy Abstract Objective To assess the role of serial FLT-PET scans during early neoadjuvant treatment as a prognostic marker of response to treatment and survival. Methods This study is a prospective cohort study which draws from a larger original study which examined the utility of FLT-PET imaging across multiple cancers. Our cohort consisted of patients who had biopsy-confirmed breast cancer amenable to surgical resection. These patients underwent serial FLT-PET scans: the first scan prior to starting neoadjuvant chemotherapy (NAC), and a second scan shortly after starting NAC. SUVmean was derived using an isocontour ROI drawn approximately half way between the SUVmax and background on three planes for each scan. The change in mean standardized uptake value (SUVmean) for the primary tumor between these two scans was then calculated, and patients were stratified into "responder" and "non-responder" groups based on a cut-off of 20% arithmetic decrease in SUVmean between the two scans. The rates of pathologic complete response (pCR) on subsequent surgical excision, overall survival (OS), and progression-free survival (PFS) were then compared between the two groups to assess for significant difference between responders and non-responders. Results 16 patients (n = 16) met criteria for inclusion and successfully underwent FLT-PET scans in the prescribed sequence of events. Seven of these patients had a decrease of 20% or larger between the two serial PET scans, making them "responders". The remaining nine patients were "non-responders" to NAC based on PET imaging. Between responders and non-responders, there was no significant difference in median PFS (7.9 years versus 3.7 years; p = 0.425) and median OS (7.5 years versus 5.0 years; p = 0.944). In the 14 patients who underwent surgical resection (n = 14), there was no significant difference in the rate of achieving pCR (33% vs. 14%; p = 0.5846) between responders and non-responders. Conclusion Further study of a larger sample size is needed to examine the potential role for FLT-PET in predicting response to neoadjuvant treatment, particularly in correlating with long-term overall and progression-free survival. Our study is limited by small sample size, but does suggest that FLT-PET has a role in the long-term prognosis of breast cancer treated with NAC and surgical resection which is worthy of further study.

123 I-FP-CIT striatal binding ratios do not decrease significantly with age in older adults Abstract Objective I-123-2β-Carbomethoxy-3β-(4-iodophenyl)-N-(3-fluoropropyl)nortropane (FP-CIT) imaging is an established biomarker used in the diagnosis of Lewy body disease. Images are often reported with the aid of striatal binding ratios (SBRs), comparing uptake to a normal database via Z scores. It is well known that SBRs are age dependent. However, previous studies cover wide age ranges between 20 and 80 years, rather than focusing on older adults. Typically a linear relationship is reported, but some authors have suggested that SBRs do not decline as rapidly in old age. Commercial software packages usually adjust the SBR Z score to attempt to compensate for age-related decline, but the model used varies. Ensuring age correction is appropriate for older adults is important, given that the majority of patients referred for FP-CIT scans are aged over 60 years. We examined the relationship of SBR with age in older adults and the effect of age correction using research scans from 123 adults over 60 years of age. Methods Twenty-nine healthy older adults and twenty-three with MCI due to Alzheimer's disease were included as controls, i.e. individuals with no evidence of Lewy body disease. Their ages ranged from 60 to 92 years (mean 76; SD 7.9). SBRs and Z scores were calculated using BRASS (Hermes Medical) and DaTQUANT (GE Healthcare). SBRs were plotted against age and linear mixed effect models applied. We tested the effect of removing age correction in BRASS using an independent dataset of 71 older adults with dementia or mild cognitive impairment. Results The slopes of the linear fits between SBR and age per year were − 0.007 (p = 0.30) with BRASS and − 0.004 (p = 0.35) with DaTQUANT. The slopes are smaller than reported in the literature and show no statistically significant difference from zero. Switching age correction off in BRASS in the test subjects reduced Z scores by approximately 1 standard deviation at 80 years of age. Conclusion We found no statistically significant age-related decline in SBR in adults over 60 years of age without Lewy body disease. Commercial software packages that apply a fixed rate of age correction may be overcorrecting for age in older adults, which could contribute to misdiagnosis.

Measurement of cerebral vascular reserves with I-123 IMP SPECT without an arterial input function using the microsphere model and radiopharmaceutical dose calibration Abstract Objectives Cerebral vascular reserve (CVR) is an important indicator for the management of and therapy for cerebral arterial occlusive disease (CAOD). Vasodilatory function is measured using the standard IMP-ARG method. The IMP autoradiography (IMP-ARG) method employed here uses a standardized input function, which was derived from 12 patients between 31 and 71 years of age. Because the population of elderly patients continues to increase in Japan, additional therapies are required to assess CVR in elderly patients with chronic cardiopulmonary disease or a history of smoking, in particular. Despite its popularity, alternatives to the IMP-ARG method are necessary. Here, we proposed the microsphere (MS) method without an input function. Method Using this method and the IMP-ARG method, we measured the CVRs of 18 CAOD patients. Results The CVRs derived with these two methods were significantly and linearly correlated (r = 0.89, p < 0.01). CVRs categorized by severity were also found to correspond between the two methods (κ = 0.87). Conclusions Thus, the method proposed here may serve as a supplemental to and be compatible with the IMP-ARG method for the assessment of CVR. Furthermore, the two methods, when used in conjunction, may result in less error than either would alone.

Improved identification of patients with oligometastatic clear cell renal cell carcinoma with PSMA-targeted 18 F-DCFPyL PET/CT Abstract Objective Complete surgical resection of metastatic sites has been shown to prolong survival in select patients with oligometastatic RCC. This treatment strategy is dependent upon the accurate characterization of a patient's extent of disease. The objective of this study was to explore the utility of PSMA-targeted 18F-DCFPyL PET/CT in patients with presumed oligometastatic clear cell RCC. Methods This is a subset analysis of a prospective study in which patients with RCC were imaged with 18F-DCFPyL PET/CT (ClinicalTrials.gov identifier NCT02687139). In the present analysis, patients with oligometastatic clear cell RCC, defined as ≤ 3 metastatic lesions on conventional imaging, were evaluated. 18F-DCFPyL PET/CT scans were reviewed for sites of disease and compared to conventional imaging. Results The final cohort included 14 patients with oligometastatic clear cell RCC. Conventional imaging revealed 21 metastatic lesions and 3 primary tumors. 18F-DCFPyL PET/CT detected 29 sites of metastatic disease and 3 primary tumors. Of the 21 metastatic lesions detected on conventional imaging, 17 (81.0%) had radiotracer uptake. Additionally, all 3 primary tumors had radiotracer uptake. In 4 (28.6%) patients a total of 12 more lesions were identified on 18F-DCFPyL PET/CT than conventional imaging. Notably, 3 (21.4%) patients were no longer considered oligometastatic. The detection rates of conventional imaging and 18F-DCFPyL PET/CT for identifying sites of disease were 66.7% and 88.9%, respectively. Conclusions PSMA-targeted PET/CT appears to aid in the identification of patients with oligometastatic clear cell RCC. If borne out in future studies, this suggests that PSMA-targeted imaging has the potential to help select candidates for metastasis-directed therapy.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Oral Biology

High CD3+ lymphocytes, low CD66b+ neutrophils, and scarce tumor budding in the invasive front of lip squamous cell carcinomas Publication date: August 2019 Source: Archives of Oral Biology, Volume 104 Author(s): Karine Duarte da Silva, Patrícia Carlos Caldeira, Alessandro Menna Alves, Ana Carolina Uchoa Vasconcelos, Ana Paula Neutzling Gomes, Maria Cássia Ferreira de Aguiar, Sandra Beatriz Chaves Tarquinio Abstract Objective This study aimed to evaluate tumor budding (TB) and quantify the neutrophilic and lymphocytic infiltration in the invasive front of lip squamous cell carcinomas. In addition, the associations between these features and the histopathological grading in the invasive front were analyzed. Design A total of 43 lip squamous cell carcinoma surgical specimens were included and classified in accordance with a histological invasive front grading system. Immunohistochemistry was performed for CD66b and CD3 for the evaluation of neutrophils and T lymphocytes, respectively, in the invasive front of the tumors. Tumor budding was evidenced by AE1/AE3 staining. Results Thirty-six (83.7%) of the tumors were well-differentiated tumors. Eleven (25.6%) of the cases exhibited high-intensity tumor budding. There were low neutrophil and high T lymphocyte infiltrations in the invasive front, leading to a low neutrophil/T lymphocyte ratio in the same region. Moreover, we found an association between tumor budding and the pattern of invasion, and between the CD3+ cell count and the inflammatory infiltrate (p < 0.05). Conclusions The low neutrophil and high T lymphocyte infiltration in the invasive front, and the few high-intensity tumor budding cases are in accordance with the histopathological features of well-differentiated lip tumors. If these characteristics remain in lip squamous cell carcinomas with more aggressive histopathological features, it deserves to be investigated.

Cannabidiol, cannabinol and their combinations act as peripheral analgesics in a rat model of myofascial pain Publication date: August 2019 Source: Archives of Oral Biology, Volume 104 Author(s): Hayes Wong, Brian E. Cairns Abstract Objective This study investigated whether local intramuscular injection of non-psychoactive cannabinoids, cannabidiol (CBD), cannabinol (CBN), cannabichromene (CBC) and their combinations can decrease nerve growth factor (NGF)-induced masticatory muscle sensitization in female rats. Design In awake rats, changes in mechanical sensitivity induced by intramuscular injection of NGF and cannabinoids were measured by applying an electronic von Frey hair over the masseter muscle to measure the withdrawal response. The effect of CBD (5 mg/ml) and CBN (1 mg/ml) or their combinations CBD/CBN (1:1 mg/ml or 5:1 mg/ml) were assessed. To confirm a peripheral action, electrophysiological experiments were undertaken in anesthetized rats to examine whether intramuscular injections of CBD (5 mg/ml) and CBN (1 mg/ml) altered the mechanical threshold of masticatory muscle mechanoreceptors. Results In behavioral experiments, CBD (5 mg/ml) or CBN (1 mg/ml) decreased NGF-induced mechanical sensitization. Combinations of CBD/CBN induced a longer-lasting reduction of mechanical sensitization than either compound alone. No significant change in mechanical withdrawal threshold was observed in the contralateral masseter muscles and no impairment of motor function was found with the inverted screen test after any of the treatments. Consistent with behavioral results, CBD (5 mg/ml), CBN (1 mg/ml) and the combination of CBD/CBN (1:1 mg/ml) increased the mechanical threshold of masseter muscle mechanoreceptors. However, combining CBD/CBN (5:1 mg/ml) at a higher ratio reduced the duration of this effect. This may indicate an inhibitory effect of higher concentrations of CBD on CBN. Conclusions These results suggest that peripheral application of these non-psychoactive cannabinoids may provide analgesic relief for chronic muscle pain disorders such as temporomandibular disorders and fibromyalgia without central side effects.

Activity and distribution pattern of enzymes in the in-situ pellicle of children Publication date: August 2019 Source: Archives of Oral Biology, Volume 104 Author(s): Susann Hertel, Annika Schulz, Roman Lang, Thomas Hofmann, Belinda König, Matthias Hannig, Christian Hannig Abstract Objective This study investigated, for the first time, pellicle enzymes with respect to their activity, distribution and fluorescence pattern in children with different caries experience. Design In-situ pellicles were collected from 41 children (aged 4–6 years) with different caries status; 17 of them were caries-free (dmf = 0), 12 had dental restorations but no current caries (dmf ≥ 2) and 12 had at least two carious lesions (dmf ≥ 2). Bovine enamel samples were fixed on individual upper jaw braces for pellicle formation. After 30 min of intraoral exposure, the pellicle and saliva samples were analysed for the activities of amylase, lysozyme, peroxidase and glucosyltransferase (GTF). The distribution of these enzymes, including GTF-isoforms B, C and D, and the pellicle ultrastructure were examined by gold-immunolabelling and transmission electron microscopy (TEM). Furthermore, interactions between pellicle enzymes and adherent bacteria were visualised using combined fluorescence and immunofluorescence labelling. Results There were no significant differences in the pellicle enzyme activities between the study groups. TEM analysis revealed the absence of GTF C and D in the pellicle of caries-active children. Amylase, peroxidase and GTF-isoforms showed a random distribution within the pellicle layer; lysozyme was found in the form of clusters. A similar ultrastructural pattern was observed for all subjects. Fluorescence labelling technique enabled visualisation of all enzymes, except for GTF B. Conclusion Pellicle enzyme activities and ultrastructure are not associated with children's caries status. Further investigation is needed to assess the influence of individual GTF-isoforms on caries susceptibility in children.

Whole metagenomic shotgun sequencing of the subgingival microbiome of diabetics and non-diabetics with different periodontal conditions Publication date: August 2019 Source: Archives of Oral Biology, Volume 104 Author(s): Roberto Farina, Mattia Severi, Alberto Carrieri, Elena Miotto, Silvia Sabbioni, Leonardo Trombelli, Chiara Scapoli Abstract Objective The aim of this study was to use high-resolution whole metagenomic shotgun sequencing to characterize the subgingival microbiome of patients with/without type 2 Diabetes Mellitus and with/without periodontitis. Design Twelve subjects, falling into one of the four study groups based on the presence/absence of poorly controlled type 2 Diabetes Mellitus and moderate-severe periodontitis, were selected. For each eligible subject, subgingival plaque samples were collected at 4 sites, all representative of the periodontal condition of the individual (i.e., non-bleeding sulci in subjects without a history of periodontitis, bleeding pockets in patients with moderate-severe periodontitis). The subgingival microbiome was evaluated using high-resolution whole metagenomic shotgun sequencing. Results The results showed that: (i) the presence of type 2 Diabetes Mellitus and/or periodontitis were associated with a tendency of the subgingival microbiome to decrease in richness and diversity; (ii) the presence of type 2 Diabetes Mellitus was not associated with significant differences in the relative abundance of one or more species in patients either with or without periodontitis; (iii) the presence of periodontitis was associated with a significantly higher relative abundance of Anaerolineaceae bacterium oral taxon 439 in type 2 Diabetes Mellitus patients. Conclusions Whole metagenomic shotgun sequencing of the subgingival microbiome was extremely effective in the detection of low-abundant taxon. Our results point out a significantly higher relative abundance of Anaerolineaceae bacterium oral taxon 439 in patients with moderate to severe periodontitis vs patients without history of periodontitis, which was maintained when the comparison was restricted to type 2 diabetics.

Applications of the oral microbiome in personalized dentistry Publication date: August 2019 Source: Archives of Oral Biology, Volume 104 Author(s): Georgios N. Belibasakis, Nagihan Bostanci, Philip D. Marsh, Egija Zaura Abstract Objective In the era of personalized medicine, it is imperative that oral health is integrated into this concept. The oral cavity fosters a highly individualized microbiome that has evolved to promote oral health, and which exists in a dynamic balance with the host. Microecological changes to the biology of the mouth [e.g. in the host diet and lifestyle, or status of the immune system] may drive deleterious shifts in the composition or metabolic activity of the oral microbiome ['dysbiosis']. This review aims to explore how knowledge of the oral microbiome may be utilized for personalized dentistry at the point-of-care. Design This is a comprehensive narrative review of the literature, summarizing the perspectives of the authors. Results The huge increase in recent knowledge on the ecology and microbiology of the oral cavity generated by 'OMIC' technologies may indeed be clinically translated to support patient care, in terms of prevention, monitoring, risk classification or early diagnosis. The identified clinical applications may not only include dental caries and periodontal disease, but also dental implants and orthodontics. Population-based applications may include systemic health, pregnancy and elderly populations. Conclusions Applications of selected oral microbiome and host-related biochemical parameters [e.g. the saliva proteome] for personalized dentistry can be customized for different clinical applications or individual populations, at point-of-care hubs.

Staircase-pattern neonatal line in human deciduous teeth is associated with tooth type Publication date: August 2019 Source: Archives of Oral Biology, Volume 104 Author(s): Jaana Hurnanen, Matti Sillanpää, Marja-Leena Mattila, Eliisa Löyttyniemi, Carsten Witzel, Jaana Rautava Abstract Objective The staircase (Sc) pattern enamel microstructure is an expression of an impaired ameloblast function. It has been reported to appear in the neonatal line (NNL), the accentuated stria evincing live birth in deciduous tooth enamel. Our objective was to investigate the prevalence of Sc NNL in deciduous tooth types and its possible association with perinatal circumstances. Design Sc in the NNL of 88 teeth, a collection derived from a long-term, prospectively followed population cohort, was recorded with linear polarised transmitted light and analysed for tooth type, duration and mode of delivery, and pain medication used during labour. Results Sc prevalence in the NNL differed highly significantly between tooth types (p < 0.0001). An increase in Sc prevalence was significantly associated with an increased ratio of prenatal to total crown enamel (p < 0.001), when buccal and lingual crown walls were analysed separately. No significant association was found between Sc prevalence and duration or mode of delivery or pain-relieving medication (p = 0.57, p = 0.65, p = 0.58, respectively). Conclusion This research indicates that the NNL location within tooth crown enamel has a strong impact on microstructural changes along the NNL. Considering our results of Sc prevalence, deciduous canines, having the least Sc appearance, could be used in studies that aim to investigate factors associated with NNL width. In addition, Sc prevalence variation in first deciduous molars might enable to investigation of physiological stressors strong enough to cause ameloblast impairment, such as Sc.

Salivary gland metabolism in an animal model of chronic kidney disease Publication date: August 2019 Source: Archives of Oral Biology, Volume 104 Author(s): R.A. Carvalho, A.C. Romero, F.K. Ibuki, F.N. Nogueira Abstract Objective The aim of this study was to determine the effects of experimental CKD into the metabolism of parotid and submandibular glands of rats. CKD was induced by 5/6 nephrectomy. Design Serum analyses of BUN (Blood Urea Nitrogen) and creatinine concentrations were performed. Major salivary glands metabolism was investigated in vivo, both at rest and during salivary stimulation conditions by NMR isotopomer analysis, using [U-13C]glucose as metabolic tracer. Results CKD increases BUN and serum creatinine concentrations (p < 0.001). Multiple metabolic alterations were detected in the parotid glands of this animal model, including decreased concentrations of alanine (p < 0.05) and creatine (p < 0.05) and increased lactate/alanine ratios (p < 0.05). The salivary stimulus fostered accumulations of acetate at both analyzed glands of the CKD model (p < 0.05), indicative of disruption of the oxidative metabolic process. Conclusions Experimental CKD induced by 5/6 nephrectomy altered the parotid salivary gland function, since glucose metabolism is clearly affected after stimulation for salivation in this gland.

Effects of Candida albicans infection on defense effector secretion by human oral mucosal epithelial cells Publication date: July 2019 Source: Archives of Oral Biology, Volume 103 Author(s): Fan Huang, Yuefeng Song, Wei Chen, Qin Liu, Qiong Wang, Weida Liu, Xiang Wang, Wenmei Wang Abstract Objective The aim of this study was to investigate the effects of Candida albicans on the production of defense effector molecules by human oral mucosal epithelial cells in vitro. Design Immortalized human oral mucosal epithelial (Leuk-1) cells and C. albicans strain 5314 were cocultured at different cell-to-C. albicans ratios. The viability of Leuk-1 cells was determined by MTT and RTCA measurements. The secretory levels of multiple defense effector molecules were determined by Enzyme-linked immunosorbent assay (ELISA). Results Our results indicated that C. albicans significantly decreased the secretion of IgG, cystatin C, lactoferrin, and TGF-β1 in a dose-dependent manner and remarkably reduced the production of IgA independent of the cell-to-C. albicans ratio. However, C. albicans clearly enhanced the secretion of IgM, galectin-3, P-selectin, granzyme B and perforin. Conclusion These results suggest that C. albicans may exert a regulatory role in the defense response of oral mucosal epithelial cells by altering secretory levels of defense effector molecules.

Liposome-coated nano doxorubicin induces apoptosis on oral squamous cell carcinoma CAL-27 cells Publication date: July 2019 Source: Archives of Oral Biology, Volume 103 Author(s): Ehab S. Abd El-Hamid, Amira M. Gamal-Eldeen, Abdelrahman M. Sharaf Eldeen Abstract Objective This study aims to investigate the apoptotic effect of Doxorubicin and its nano-formulated form (Doxil) on oral squamous cell carcinoma CAL-27 cells. Design Cell viability using MTT assay, mode of cell death using fluorescence analysis, expression of the apoptotic marker caspase-3 using indirect ELISA technique and expression of C-Myc gene using reverse transcriptase and real time PCR. Results Doxil treatment resulted in a higher percentage of apoptotic cells than doxorubicin treatment, while doxorubicin treatment resulted in a higher percentage of necrotic cells than Doxil treatment. Doxil-treated cells exhibited 3.38-fold higher caspase-3 levels than control cells, while doxorubicin significantly increased caspase-3 levels by 2.72-fold. The percentage of C-Myc mRNA inhibition was 27% in doxorubicin-treated cells and 41% in Doxil-treated cells. Conclusions Doxil exerted a higher apoptotic effect on CAL-27 cells compared to doxorubicin. It showed a higher increase in capase-3 level than doxorubicin and also exerted a more percentage of C-Myc mRNA inhibition.

BMP4 mutations in tooth agenesis and low bone mass Publication date: July 2019 Source: Archives of Oral Biology, Volume 103 Author(s): Miao Yu, Hao Wang, Zhuangzhuang Fan, Chencheng Xie, Haochen Liu, Yang Liu, Dong Han, Sing-Wai Wong, Hailan Feng Abstract Objective To identify an uncommon genetic cause of tooth agenesis (TA) by utilizing whole exome sequencing (WES) and targeted Sanger sequencing in a cohort of 120 patients with isolated TA. Design One deleterious mutation in the gene encoding bone morphogenetic protein 4 (BMP4) was identified in 6 unrelated patients with TA by WES. After that, the coding exons of BMP4 were examined in 114 TA patients using Sanger sequencing. Dual-energy X-ray absorptiometry (DEXA) was used to measure the bone mineral density of patients who carried a BMP4 mutation. Finally, preliminary functional studies of two BMP4 mutants were performed. Results We detected 3 novel missense mutations (c.58 G > A: p.Gly20Ser, c.326 G > T: p.Arg109Leu and c.614 T > C: p.Val205Ala) and 1 reported mutation in the BMP4 gene among 120 TA probands. The previously reported BMP4 mutation (c.751C > T: p.His251Tyr) was associated with urethra and eye anomalies. By extending the pedigrees, we determined that the tooth phenotypes had an autosomal dominant inheritance pattern, as individuals carrying a BMP4 mutation exhibit different types of dental anomalies. Interestingly, we observed that patients harboring a BMP4 mutation manifested early onset osteopenia or osteoporosis. Further in vitrofunctional assays demonstrated that two BMP4 mutants resulted in a decreased activation of Smad signaling. Therefore, a loss-of-function in BMP4 may contribute to the clinical phenotypes seen in this study. Conclusions We identified 4 mutations in the BMP4 gene in 120 TA patients. To our knowledge, this is the first study to describe human skeletal diseases associated with BMP4 mutations.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Computer Assisted Radiology and Surgery

EasyLabels: weak labels for scene segmentation in laparoscopic videos Abstract Purpose We present a different approach for annotating laparoscopic images for segmentation in a weak fashion and experimentally prove that its accuracy when trained with partial cross-entropy is close to that obtained with fully supervised approaches. Methods We propose an approach that relies on weak annotations provided as stripes over the different objects in the image and partial cross-entropy as the loss function of a fully convolutional neural network to obtain a dense pixel-level prediction map. Results We validate our method on three different datasets, providing qualitative results for all of them and quantitative results for two of them. The experiments show that our approach is able to obtain at least \(90\%\) of the accuracy obtained with fully supervised methods for all the tested datasets, while requiring \(\sim 13\) \(\times \) less time to create the annotations compared to full supervision. Conclusions With this work, we demonstrate that laparoscopic data can be segmented using very few annotated data while maintaining levels of accuracy comparable to those obtained with full supervision.

Position-based modeling of lesion displacement in ultrasound-guided breast biopsy Abstract Purpose Although ultrasound (US) images represent the most popular modality for guiding breast biopsy, malignant regions are often missed by sonography, thus preventing accurate lesion localization which is essential for a successful procedure. Biomechanical models can support the localization of suspicious areas identified on a preoperative image during US scanning since they are able to account for anatomical deformations resulting from US probe pressure. We propose a deformation model which relies on position-based dynamics (PBD) approach to predict the displacement of internal targets induced by probe interaction during US acquisition. Methods The PBD implementation available in NVIDIA FleX is exploited to create an anatomical model capable of deforming online. Simulation parameters are initialized on a calibration phantom under different levels of probe-induced deformations; then, they are fine-tuned by minimizing the localization error of a US–visible landmark of a realistic breast phantom. The updated model is used to estimate the displacement of other internal lesions due to probe-tissue interaction. Results The localization error obtained when applying the PBD model remains below 11 mm for all the tumors even for input displacements in the order of 30 mm. This proposed method obtains results aligned with FE models with faster computational performance, suitable for real-time applications. In addition, it outperforms rigid model used to track lesion position in US-guided breast biopsies, at least halving the localization error for all the displacement ranges considered. Conclusion Position-based dynamics approach has proved to be successful in modeling breast tissue deformations during US acquisition. Its stability, accuracy and real-time performance make such model suitable for tracking lesions displacement during US-guided breast biopsy.

IJCARS—IPCAI 2019 special issue: conference information processing for computer-assisted interventions, 10th international conference 2019—part 1

Automatic self-gated 4D-MRI construction from free-breathing 2D acquisitions applied on liver images Abstract Purpose MRI slice reordering is a necessary step when three-dimensional (3D) motion of an anatomical region of interest has to be extracted from multiple two-dimensional (2D) dynamic acquisition planes, e.g., for the construction of motion models used for image-guided radiotherapy. Existing reordering methods focus on obtaining a spatially coherent reconstructed volume for each time. However, little attention has been paid to the temporal coherence of the reconstructed volumes, which is of primary importance for accurate 3D motion extraction. This paper proposes a fully automatic self-sorting four-dimensional MR volume construction method that ensures the temporal coherence of the results. Methods First, a pseudo-navigator signal is extracted for each 2D dynamic slice acquisition series. Then, a weighted graph is created using both spatial and motion information provided by the pseudo-navigator. The volume at a given time point is reconstructed following the shortest paths in the graph starting that time point of a reference slice chosen based on its pseudo-navigator signal. Results The proposed method is evaluated against two state-of-the-art slice reordering algorithms on a prospective dataset of 12 volunteers using both spatial and temporal quality metrics. The automated end-exhale extraction showed results closed to the median value of the manual operators. Furthermore, the results of the validation metrics show that the proposed method outperforms state-of-the-art methods in terms of both spatial and temporal quality. Conclusion Our approach is able to automatically detect the end-exhale phases within one given anatomical position and cope with irregular breathing.

Uncertainty-aware performance assessment of optical imaging modalities with invertible neural networks Abstract Purpose Optical imaging is evolving as a key technique for advanced sensing in the operating room. Recent research has shown that machine learning algorithms can be used to address the inverse problem of converting pixel-wise multispectral reflectance measurements to underlying tissue parameters, such as oxygenation. Assessment of the specific hardware used in conjunction with such algorithms, however, has not properly addressed the possibility that the problem may be ill-posed. Methods We present a novel approach to the assessment of optical imaging modalities, which is sensitive to the different types of uncertainties that may occur when inferring tissue parameters. Based on the concept of invertible neural networks, our framework goes beyond point estimates and maps each multispectral measurement to a full posterior probability distribution which is capable of representing ambiguity in the solution via multiple modes. Performance metrics for a hardware setup can then be computed from the characteristics of the posteriors. Results Application of the assessment framework to the specific use case of camera selection for physiological parameter estimation yields the following insights: (1) estimation of tissue oxygenation from multispectral images is a well-posed problem, while (2) blood volume fraction may not be recovered without ambiguity. (3) In general, ambiguity may be reduced by increasing the number of spectral bands in the camera. Conclusion Our method could help to optimize optical camera design in an application-specific manner.

Weakly supervised convolutional LSTM approach for tool tracking in laparoscopic videos Abstract Purpose Real-time surgical tool tracking is a core component of the future intelligent operating room (OR), because it is highly instrumental to analyze and understand the surgical activities. Current methods for surgical tool tracking in videos need to be trained on data in which the spatial positions of the tools are manually annotated. Generating such training data is difficult and time-consuming. Instead, we propose to use solely binary presence annotations to train a tool tracker for laparoscopic videos. Methods The proposed approach is composed of a CNN + Convolutional LSTM (ConvLSTM) neural network trained end to end, but weakly supervised on tool binary presence labels only. We use the ConvLSTM to model the temporal dependencies in the motion of the surgical tools and leverage its spatiotemporal ability to smooth the class peak activations in the localization heat maps (Lh-maps). Results We build a baseline tracker on top of the CNN model and demonstrate that our approach based on the ConvLSTM outperforms the baseline in tool presence detection, spatial localization, and motion tracking by over \(5.0\%\) , \(13.9\%\) , and \(12.6\%\) , respectively. Conclusions In this paper, we demonstrate that binary presence labels are sufficient for training a deep learning tracking model using our proposed method. We also show that the ConvLSTM can leverage the spatiotemporal coherence of consecutive image frames across a surgical video to improve tool presence detection, spatial localization, and motion tracking.

Deformable multimodal registration for navigation in beating-heart cardiac surgery Abstract Purpose: Minimally invasive beating-heart surgery is currently performed using endoscopes and without navigation. Registration of intraoperative ultrasound to a preoperative cardiac CT scan is a valuable step toward image-guided navigation. Methods: The registration was achieved by first extracting a representative point set from each ultrasound image in the sequence using a deformable registration. A template shape representing the cardiac chambers was deformed through a hierarchy of affine transformations to match each ultrasound image using a generalized expectation maximization algorithm. These extracted point sets were matched to the CT by exhaustively searching over a large number of precomputed slices of 3D geometry. The result is a similarity transformation mapping the intraoperative ultrasound to preoperative CT. Results: Complete data sets were acquired for four patients. Transesophageal echocardiography ultrasound sequences were deformably registered to a model of oriented points with a mean error of 2.3 mm. Ultrasound and CT scans were registered to a mean of 3 mm, which is comparable to the error of 2.8 mm expected by merging ultrasound registration with uncertainty of cardiac CT. Conclusion: The proposed algorithm registered 3D CT with dynamic 2D intraoperative imaging. The algorithm aligned the images in both space and time, needing neither dynamic CT imaging nor intraoperative electrocardiograms. The accuracy was sufficient for navigation in thoracoscopically guided beating-heart surgery.

Catheter localization in 3D ultrasound using voxel-of-interest-based ConvNets for cardiac intervention Abstract Purpose Efficient image-based catheter localization in 3D US during cardiac interventions is highly desired, since it facilitates the operation procedure, reduces the patient risk and improves the outcome. Current image-based catheter localization methods are not efficient or accurate enough for real clinical use. Methods We propose a catheter localization method for 3D cardiac ultrasound (US). The catheter candidate voxels are first pre-selected by the Frangi vesselness filter with adaptive thresholding, after which a triplanar-based ConvNet is applied to classify the remaining voxels as catheter or not. We propose a Share-ConvNet for 3D US, which reduces the computation complexity by sharing a single ConvNet for all orthogonal slices. To boost the performance of ConvNet, we also employ two-stage training with weighted cross-entropy. Using the classified voxels, the catheter is localized by a model fitting algorithm. Results To validate our method, we have collected challenging ex vivo datasets. Extensive experiments show that the proposed method outperforms state-of-the-art methods and can localize the catheter with an average error of 2.1 mm in around 10 s per volume. Conclusion Our method can automatically localize the cardiac catheter in challenging 3D cardiac US images. The efficiency and accuracy localization of the proposed method are considered promising for catheter detection and localization during clinical interventions.

Toward an automatic preoperative pipeline for image-guided temporal bone surgery Abstract Purpose Minimally invasive surgery is often built upon a time-consuming preoperative step consisting of segmentation and trajectory planning. At the temporal bone, a complete automation of these two tasks might lead to faster interventions and more reproducible results, benefiting clinical workflow and patient health. Methods We propose an automatic segmentation and trajectory planning pipeline for image-guided interventions at the temporal bone. For segmentation, we use a shape regularized deep learning approach that is capable of automatically detecting even the cluttered tiny structures specific for this anatomy. We then perform trajectory planning for both linear and nonlinear interventions on these automatically segmented risk structures. Results We evaluate the usability of segmentation algorithms for planning access canals to the cochlea and the internal auditory canal on 24 CT data sets of real patients. Our new approach achieves similar results to the existing semiautomatic method in terms of Dice but provides more accurate organ shapes for the subsequent trajectory planning step. The source code of the algorithms is publicly available. Conclusion Automatic segmentation and trajectory planning for various clinical procedures at the temporal bone are feasible. The proposed automatic pipeline leads to an efficient and unbiased workflow for preoperative planning.

Face detection in the operating room: comparison of state-of-the-art methods and a self-supervised approach Abstract Purpose Face detection is a needed component for the automatic analysis and assistance of human activities during surgical procedures. Efficient face detection algorithms can indeed help to detect and identify the persons present in the room and also be used to automatically anonymize the data. However, current algorithms trained on natural images do not generalize well to the operating room (OR) images. In this work, we provide a comparison of state-of-the-art face detectors on OR data and also present an approach to train a face detector for the OR by exploiting non-annotated OR images. Methods We propose a comparison of six state-of-the-art face detectors on clinical data using multi-view OR faces, a dataset of OR images capturing real surgical activities. We then propose to use self-supervision, a domain adaptation method, for the task of face detection in the OR. The approach makes use of non-annotated images to fine-tune a state-of-the-art detector for the OR without using any human supervision. Results The results show that the best model, namely the tiny face detector, yields an average precision of 0.556 at intersection over union of 0.5. Our self-supervised model using non-annotated clinical data outperforms this result by 9.2%. Conclusion We present the first comparison of state-of-the-art face detectors on OR images and show that results can be significantly improved by using self-supervision on non-annotated data.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Microbiology

Flavobacterium viscosus sp. nov. and Flavobacterium tangerina sp. nov., from Primates Feces Abstract Strains YIM 102796T and YIM 102701-2T were isolated from the feces of Macaca mulatta and Hylobates hoolock, respectively, living in the Yunnan Wild Animal Park, Yunnan province of China. The two strains were Gram-stain-negative, non-gliding, produced flexirubin pigments, non-flagellated and aerobic bacteria. The 16S rRNA gene-based phylogenetic analysis indicate that both YIM 102796T and YIM 102701-2T are members of the genus Flavobacterium, closely related to F. ummariense DS-12T (95.9% similarity) and F. ceti 454-2T (93.8% similarity), respectively. The two strains shared 95.1 % 16S rRNA gene sequence similarity. The average nucleotide identity and digital DNA–DNA hybridization values between the two strains were 76.5% and 22.9%, respectively, indicating that they are separate species. DNA G+C contents of YIM 102796T and YIM 102701-2Twere 32.3 mol% and 34.0 mol%, respectively. Strains are able to grow at 4–37 °C, at pH 7.0–8.0 and in 0–2% (w/v) NaCl. Predominant fatty acid constituents (>7 %) were iso-C15:0, summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c) and summed feature 9 (iso-C17:1ω9c and/or 10-methylC16:0). Menaquinone 6 is major respiratory quinone. The predominant polar lipids were very similar to each other, comprising phosphatidylethanolamine, and multiple unknown aminolipids and unidentified polar lipids, and an unidentified aminophospholipid. On the basis of phenotypic and phylogenetic distinctiveness, it is suggested that the two strains represent two novel Flavobacterium species with strain YIM 102796T (=KCTC 52101T=CCTCC AB 2016015T) as the type strain of Flavobacterium viscosus sp. nov. and strain YIM 102701-2T (=KCTC 52100T=CCTCC AB 2016028T) as the type strain of Flavobacterium tangerina sp. nov.

Comparison of Aspergillus Section Nigri Species Populations in Conventional and Organic Raisin Vineyards Abstract Species belonging to Aspergillus section Nigri are widespread in the vineyard environment, both in soil and on plant surfaces. We used plate counts and droplet digital PCR (ddPCR) methods to compare populations of the four most prevalent species (A. carbonarius, A. niger, A. welwitschiae, and A. tubingensis) over two consecutive years in conventional and organic vineyards, to determine whether management affects the potential distribution of mycotoxigenic Aspergillus species. In 2016, plate counts showed that soil populations of total filamentous fungi and of Aspergillus section Nigri species were not significantly different between conventional and organic vineyards. In 2017, while total fungal populations in soil were not significantly different, Aspergillus section Nigri populations were significantly higher in organic vineyard soil. In both years, there were no significant differences in total fungal populations and in Aspergillus section Nigri populations on fruit surfaces collected from conventional and organic vineyards. Likewise, ddPCR methods did not show significant differences in percent distribution of Aspergillus species in soil and fruit between conventional and organic vineyards. These results suggest that intervention strategies for preharvest control of potential mycotoxigenic fungi are likely to be equally compatible with either vineyard management strategy.

Rapid Detection of Phosphate-Solubilizing Bacteria from Agricultural Areas in Erzurum Abstract In this study, the newly designed pqq gene-specific primer sets were used for determination of phosphate-solubilizing capabilities of bacterial isolates from the agricultural regions of Erzurum. The specificity of newly designed primer sets (PqqA2F/PqqA2R, Pqq5F/Pqq5R, PqqF2/PqqF2R) were tested against ten isolates, whose phosphate-solubilizing activities were initially proved by the conventional methods. Non-phosphate-solubilizing bacteria were also chosen as negative control. According to the results, five of ten phosphate-solubilizing bacteria with PqqA2F/PqqA2R, two of ten phosphate-solubilizing bacteria with Pqq5F/Pqq5R primer set, and one of ten phosphate solubilizing with PqqF2F/PqqF2R bacteria were successfully amplificated in the PCR assay and none of the non-phosphate-solubilizing bacteria was amplificated. Then, the molecular characterization of the active phosphate-solubilizing strains was done based on the partial 16S ribosomal RNA gene region sequence analysis method. Two isolates of Enterobacter sp., 1 Rhizobium sp., 1 Enterococcussp., 1 Bacillus cereus, 1 Bacillus atrophaeus, 1 Bacillus aryabhattai, 1 Acinetobacter sp., 1 Pseudomonas japonica, and 1 Enterobacter cloacae were identified as active phosphate-solubilizing strains. Consequently, the results showed that this specific primer sets could be used as an economic, rapid, and useful tool for the detection of phosphate-solubilizing strains in the agricultural researches.

Streptococcus periodonticum sp. nov., Isolated from Human Subgingival Dental Plaque of Periodontitis Lesion Abstract A novel facultative anaerobic and Gram-stain-positive coccus, designated strain ChDC F135T, was isolated from human subgingival dental plaque of periodontitis lesion and was characterized by polyphasic taxonomic analysis. The 16S rRNA gene (16S rDNA) sequence of strain ChDC F135T was closest to that of Streptococcus sinensis HKU4T (98.2%), followed by Streptococcus intermedia SK54T (97.0%), Streptococcus constellatusNCTC11325T (96.0%), and Streptococcus anginosus NCTC 10713T (95.7%). In contrast, phylogenetic analysis based on the superoxide dismutase gene (sodA) and the RNA polymerase beta-subunit gene (rpoB) showed that the nucleotide sequence similarities of strain ChDC F135T were highly similar to the corresponding genes of S. anginosus NCTC 10713T (99.2% and 97.6%, respectively), S. constellatus NCTC11325T (87.8% and 91.4%, respectively), and S. intermedia SK54T (85.8% and 91.2%, respectively) rather than those of S. sinensis HKU4T (80.5% and 82.6%). The complete genome of strain ChDC F135T consisted of 1,901,251 bp and the G+C content was 38.9 mol %. Average nucleotide identity value between strain ChDC F135T and S. sinensis HKU4T or S. anginosus NCTC 10713T were 75.7% and 95.6%, respectively. The C14:0 composition of the cellular fatty acids of strain ChDC F135T (32.8%) was different from that of S. intermedia (6–8%), S. constellatus (6–13%), and S. anginosus (13–20%). Based on the results of phylogenetic and phenotypic analysis, strain ChDC F135T (= KCOM 2412T = JCM 33300T) was classified as a type strain of a novel species of the genus Streptococcus, for which we proposed the name Streptococcus periodonticum sp. nov.

The OmpA Gene of Xanthomonas axonopodis pv. glycines is Involved in Pathogenesis of Pustule Disease on Soybean Abstract The goal of this study was to elucidate the role of the outer membrane protein A (ompA) gene of Xanthomonas axonopodis pv. glycines in bacterial pustule pathogenesis of soybean. An ompA mutant of X. axonopodis pv. glycines KU-P-SW005 was shown to significantly decrease cellulase, pectate lyase, and polysaccharide production. The production of these proteins in the ompA mutant was approximately five times lower than that of the wildtype. The ompA mutant also exhibited modified biofilm development. More importantly, the mutant reduced disease severity to the soybean. Ten days after inoculation, the virulence rating of the susceptible soybean cv. SJ4 inoculated with the ompA mutant was 11.23%, compared with 87.98% for the complemented ompA mutant. Production of cellulase, pectate lyase, polysaccharide was restored, biofilm, and pustule numbers were restored in the complemented ompA mutant that did not differ from the wild type. Taken together, these data suggest that OmpA-mediated invasion plays an important role in protein secretion during pathogenesis to soybean.

Secondary Metabolites and PI3K Inhibitory Activity of Colletotrichum gloeosporioides , a Fungal Endophyte of Uncaria rhynchophylla Abstract In the present study, nine compounds (1–9) were isolated from Colletotrichum gloeosporioides (an endophytic fungus from Uncaria rhynchophylla) which was cultured in wheat bran medium. Their structures were elucidated as 4-Epi-14-hydroxy-10, 23-dihydro-24, 25-dehydroaflavinine (1), 10, 23-Dihydro-24,25 -dehydro-21–oxoaflavinine (2), Ergosterol (3), Ergosterol peroxide (4), Mellein (5), 4, 5-dihydroblumenol A (6), Colletotrichine A (7), Cyclo(L-leucyl-L-leucyl) (8), and Brevianamide F (9) based on NMR spectral data, as well as comparing with previous literature data. This is the first report about the isolation of compounds 1–2, 6, and 8–9 from Colletotrichum genus. All compounds were tested for their phosphoinositide 3-kinase (PI3Kα) inhibitory activity. Compounds 8 and 9 showed potent PI3K α inhibitory activity with IC50 values of 38.1 and 4.8 µM, respectively, while the other compounds showed very weak activity at a concentration of 20 µg/mL.

Microbial Diversity in the Edible Gall on White Bamboo Formed by the Interaction between Ustilago esculenta and Zizania latifolia Abstract An edible gall is formed between the third and fourth nodes beneath the apical meristem near the base of Zizania latifolia shoots. This gall is harbored by and interacts with the smut fungus Ustilago esculenta. The gall is also a valuable vegetable called "white bamboo," jiaobai or gausun in China and makomotake in Japan. Five samples of the galls harvested at different stages of swelling were used to isolate microorganisms by culturing. Isolated fungal and bacterial colonies were identified by DNA sequencing and matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry, respectively. Several strains of U. esculenta as well as 6 other species of fungi and 10 species of bacteria were isolated. The microbiome was also evaluated by simple and outlined DNA profiling with automated rRNA intergenic spacer analysis (ARISA), and the amount of DNA of U. esculenta was determined by qPCR. At least 16 species of fungi and 40 species of bacteria were confirmed by ARISA of the overall sample. Interestingly, the greatest bacterial diversity, i.e., 18 species, was observed in the most mature sample, whereas the fungal diversity observed in this sample, i.e., 4 species, was rather poor. Based on qPCR, U. esculenta occurred in samples from all stages; however, the abundance of U. esculentaexhibited unique U-shaped relationships with growth. These results may explain why the interaction between U. esculenta and Z. latifolia also influences the unique microbial diversity observed throughout the growth stages of the swollen shoot, although the limited sample size does not allow conclusive findings.

Molecular Diversity of Cyanopodoviruses in Two Coastal Wetlands in Northeast China Abstract Although bacteriophages are the most abundant biological entities on the planet, their genetic diversity, especially in natural wetlands, is poorly understood. In this study, the genetic diversity of cyanopodoviruses in sediments of two coastal wetlands in Northeast China was investigated by targeting the DNA polymerase (pol) gene. A total of 66 DNA pol clones were obtained. A BLAST search at the amino acid level showed that the obtained sequences had the highest identity ranged from 83 to 99% to the known sequences. A phylogenetic tree showed that the distribution patterns of DNA pol sequence were different between two wetland soils, and 29 clones of this study formed four wetland-specific groups, which suggested that unrevealed novel groups of cyanopodovirus inhabited in wetlands. In addition, nonmetric multidimensional scaling (NMDS) analysis of all DNA pol sequences from various environments showed that cyanopodovirus communities of coastal wetlands are in the intermediate position between marine water environments and terrestrial freshwater environments, which highlights that the coastal wetlands as transitional zones between inland freshwater environments and marine environments.

High-Throughput Sequencing Reveals the Gut Microbiome of the Bactrian Camel in Different Ages Abstract The complex gut microbiota plays a key role in host metabolism and health. However, the core microbial communities in the different aged Bactrian camels remain totally unclear. We used high-throughput 16S rRNA gene sequencing to examine the temporal variability of the fecal microbiota in Bactrian camels. At 2 months of age, the fecal microbiota was composed of Firmicutes, Proteobacteria, and Actinobacteria. At 1 and 3 years of age, the fecal microbiota was dominated by Firmicutes, Bacteroidetes, and Verrucomicrobia. At the genus level, Blautia, Fusobacterium, and Bifidobacterium were more abundant at 2 months of age, as well as Escherichia–Shigella. Ruminococcaceae_UCG-005, Akkermansia, and Christensenellaceae_R-7_group were the most abundant at 1 and 3 years of age. Diversity and stability of the gut microbiota increased with age. There was enrichment for genes associated with immune system diseases at 2 months of age. This study is the first to investigate the distribution of the gut microbiota in Bactrian camels with different ages and provide a baseline for future camel microbiology research.

A Study on Acinetobacter baumannii and Staphylococcus aureus Strains Recovered from the Same Infection Site of a Diabetic Patient Abstract Diabetic foot ulcer infections are frequently polymicrobial in nature and exhibit increased morbidity and mortality, as well as, treatment failures. Interactions between Acinetobacter baumannii and Staphylococcus aureuswere studied, which showed strain-dependent changes in growth and antibiotic susceptibility. This study examined the interactions between two clinical strains of A. baumannii (1929) and S. aureus (1928) that were recovered from skin and soft tissues of a diabetic patient. When S. aureus 1928 and A. baumannii 1929 were co-cultured together, there was no significant decrease in growth in either clinical strains, indicating that both strains can co-exist in the same site of infection. Additionally, neither strains experienced statistically significant changes in susceptibility. These findings highlight that these two pathogens can be found in the same niche of infection, which may lead to more aggressive outcome of the infection.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com