Draft genome sequences of clinical Mycobacterium canettii strains in Canada.

Mycobacterium canettii is a rare pathogen causing tuberculosis in humans and presents a risk to public health. Here, we report the genome sequences of two M. canettii strains. The genomes will assist in creating sequence-based tools for M. canettii and serve as references for identification, surveillance, and epidemiological investigations.

Prognostic Nomogram, Demographics and Comparative Analysis of Urinary Bladder Small Cell and Large Cell Neuroendocrine Carcinoma.

BACKGROUND: This retrospective study aims to provide a comprehensive analysis of the demographics, survival rates, and therapeutic approaches of small-cell neuroendocrine carcinoma (SCNEC) and large-cell neuroendocrine carcinoma (LCNEC) while highlighting key differences compared to common urinary bladder cancers. METHODS: Our analysis utilized the Surveillance, Epidemiology, and End Results database (SEER), and data was collected from 2000-2020. RESULTS: A total of 1040 cases of urinary bladder SCNEC and LCNEC were identified. Most patients were over the age of 80 years (33.2%), male (78.9%), and Caucasian (83.6%). Most tumors were over 4.1cm (47.4%) and in the lateral wall of the bladder (37.8%). The overall 5-year survival was 22.1% (95% confidence interval (95% CI):20.7-23.5). The 5-year survival by sex was greatest for the female population (28.0%; (95% CI: 24.5-35.0). For treatment modality, the 5-year survival for each was as follows: surgery, 12.5% (95% CI: 10.5-14.5) multimodality therapy (surgery and chemotherapy), 31.1% (95% CI: 28.5-33.7) and combination (surgery, chemotherapy, and radiation) 32.8% (95% CI: 29.1-36.5). On multivariable analysis, positive nodal status hazar ratio (HR)(HR3.65 [95% CI: 2.34-5.71], P < .001) was identified as a negative predictor for survival, and increasing age was nearly significant for a worse prognosis (P = .052). The prognostic nomogram that was created to predict patient survivability mirrored the findings from the statistical analysis, with a statistically significant difference found in race, treatment modality, and tumor stage. CONCLUSIONS: SCNEC and LCNEC are rare yet highly intrusive subtypes of bladder cancer that usually affect Caucasian males over the age of 80 years old. The study identifies older age and positive nodal status as adverse prognostic indicators. Our findings offer crucial insights that can inform future clinical guidelines and serve as a basis for more tailored treatment strategies for these aggressive subtypes of bladder cancer.

Successful endoscopic submucosal dissection of an esophageal granular cell tumor in a pediatric patient: A case report and a therapeutic insight.

Esophageal granular cell tumors (GCTs) are rare mesenchymal neoplasms that originate from the Schwann cells of the neural sheath in the esophageal wall. Esophageal GCTs represent approximately 2% of all GCTs. Most cases of esophageal GCT occur in adults with few cases reported in pediatric patients. Although typically benign, these tumors can occasionally exhibit malignant behavior, necessitating timely and appropriate intervention. Traditionally, surgical resection was considered for treatment, nonetheless, considering the invasive nature of such interventions, endoscopic approaches have been developed for diagnosis and treatment. Endoscopic approaches have been shown to lead to serious complications at times, such as incomplete resection or perforation. Here, we present a successful application of endoscopic submucosal dissection in the treatment of an adolescent female patient with esophageal GCT which was discovered during her prior esophagogastroduodenoscopy for vomiting.

Mutations in embB406 Are Associated with Low-Level Ethambutol Resistance in Canadian Mycobacterium tuberculosis Isolates.

In Mycobacterium tuberculosis, molecular predictions of ethambutol resistance rely primarily on the detection of mutations within embB. However, discordance between embB406 mutations and gold standard phenotypic drug sensitivity testing (DST) questions the significance of embB406 mutations used in molecular DST. This study tabulates embB mutations found in Canadian M. tuberculosis isolates and evaluates the impact of specific mutations on ethambutol resistance. The National Reference Centre for Mycobacteriology culture collection (n = 2796) was screened for isolates with embB mutations. Phenotypic DST was performed on the BACTEC™ MGIT™ 960 at ethambutol concentrations of 2-5 µg/mL. Whole genome sequencing was used for drug resistance predictions, phylogenomics and single nucleotide polymorphism analysis. Detection of resistance-associated embB mutations corresponded to a positive predictive value of 64.3%, negative predictive value of 99.2%, 98.7% specificity, and 73.3% sensitivity compared to phenotypic DST. Two embB406 mutation subtypes (Gly406Asp, Gly406Ala) were found among 16 isolates, of which 12 were sensitive at 5 µg/mL ethambutol with variable resistance between 2-4 µg/mL. A novel frameshift mutation in regulator embR (Gln258fs) was found in nine isolates. Mutations in embB406 were associated with low-level ethambutol resistance undetectable at the recommended critical concentration (5 µg/mL). These novel mutations may exacerbate variability in ethambutol resistance.

Evaluation of Whole Genome Sequencing-Based Predictions of Antimicrobial Resistance to TB First Line Agents: A Lesson from 5 Years of Data.

Phenotypic susceptibility testing of the Mycobacterium tuberculosis complex (MTBC) isolate requires culture growth, which can delay rapid detection of resistant cases. Whole genome sequencing (WGS) and data analysis pipelines can assist in predicting resistance to antimicrobials used in the treatment of tuberculosis (TB). This study compared phenotypic susceptibility testing results and WGS-based predictions of antimicrobial resistance (AMR) to four first-line antimicrobials-isoniazid, rifampin, ethambutol, and pyrazinamide-for MTBC isolates tested between the years 2018-2022. For this 5-year retrospective analysis, the WGS sensitivity for predicting resistance for isoniazid, rifampin, ethambutol, and pyrazinamide using Mykrobe was 86.7%, 100.0%, 100.0%, and 47.8%, respectively, and the specificity was 99.4%, 99.5%, 98.7%, and 99.9%, respectively. The predictive values improved slightly using Mykrobe corrections applied using TB Profiler, i.e., the WGS sensitivity for isoniazid, rifampin, ethambutol, and pyrazinamide was 92.31%, 100%, 100%, and 57.78%, respectively, and the specificity was 99.63%. 99.45%, 98.93%, and 99.93%, respectively. The utilization of WGS-based testing addresses concerns regarding test turnaround time and enables analysis for MTBC member identification, antimicrobial resistance prediction, detection of mixed cultures, and strain genotyping, all through a single laboratory test. WGS enables rapid resistance detection compared to traditional phenotypic susceptibility testing methods using the WHO TB mutation catalog, providing an insight into lesser-known mutations, which should be added to prediction databases as high-confidence mutations are recognized. The WGS-based methods can support TB elimination efforts in Canada and globally by ensuring the early start of appropriate treatment, rapidly limiting the spread of TB outbreaks.

Characterization of Mycobacterium orygis, Mycobacterium bovis, and Mycobacterium caprae Infections in Humans in Western Canada.

Epidemiologic research on zoonotic tuberculosis historically used Mycobacterium bovis as a surrogate measure; however, increased reports of human tuberculosis caused by other animal-associated Mycobacterium tuberculosis complex members like Mycobacterium orygis necessitates their inclusion. We performed a retrospective cohort study including persons infected with any animal-lineage M tuberculosis complex species in Alberta, Canada, from January 1995 to July 2021, identifying 42 patients (20 M bovis, 21 M orygis, 1 M caprae). Demographic, epidemiologic, and clinical characteristics were compared against persons with culture-confirmed M tuberculosis infection. The proportion of culture-positive infections caused by M orygis increased continuously from 2016 to 2020. Significantly more females at a higher median age were impacted by M orygis, with all patients originating from South Asia. Mycobacterium bovis caused significantly more extrapulmonary disease and disproportionately impacted young females, particularly those pregnant or postpartum. All infections were acquired abroad. These findings can aid in developing targeted public health interventions.

Continuity obstetric care demonstrates greater vaginal birth after caesarean success.

BACKGROUND: Australia's caesarean rate is higher than Organisation for Economic Co-operation and Development (OECD) average, and is rising. Vaginal birth after caesarean (VBAC) is safe for selected women. Midwifery continuity of care (CoC) is associated with higher rates of vaginal birth compared to other models; however, impacts on VBAC attempts and success are unknown. AIMS: The primary aim was to determine if there is a difference in achieving VBAC between CoC and non-CoC (NCoC) models. The secondary aim was to determine if there is a difference in the proportion of women attempting VBAC between these models. MATERIALS AND METHODS: Retrospective review of antenatal records and birthing data of all women who birthed in 2021 with one or more previous caesareans. Women were included if they had two or fewer caesareans. Women were excluded if contraindications to VBAC existed. RESULTS: There were 142/1109 (12.8%) women who had previous caesareans and were eligible to attempt VBAC. There were 47/109 (43.1%) women who attempted vaginal birth after one caesarean with 78.7% success. After one caesarean, women in CoC were more likely to achieve VBAC than NCoC (45.2% vs 26.1%; relative risk (RR) 1.76, 95% CI 1.04-3.00), although when stratified by private and midwifery CoC models, women in midwifery CoC models were more likely to be successful (private RR 0.69, 95% CI 0.23-2.07 vs midwifery RR 2.48, 95% CI 1.50-4.11). Women in CoC were more likely to attempt VBAC (54.7% vs 34.8%; RR 1.57, 95% CI 1.02-2.41), and receive counselling about VBAC (92.5% vs 62%; RR 1.48, 95% CI 1.41-3.11). CONCLUSION: CoC improves the rate of attempted and successful VBAC through several factors, including increased counselling and greater provision of birth choices.

Whole genome sequencing-based identification of human tuberculosis caused by animal-lineage Mycobacterium orygis.

A recently described member of the Mycobacterium tuberculosis complex (MTBC) is Mycobacterium orygis, which can cause disease primarily in animals but also in humans. Although M. orygis has been reported from different geographic regions around the world, due to a lack of proper identification techniques, the contribution of this emerging pathogen to the global burden of zoonotic tuberculosis is not fully understood. In the present work, we report single nucleotide polymorphism (SNP) analysis using whole genome sequencing (WGS) that can accurately identify M. orygis and differentiate it from other members of the MTBC species. WGS-based SNP analysis was performed for 61 isolates from different provinces in Canada that were identified as M. orygis. A total of 56 M. orygis sequences from the public databases were also included in the analysis. Several unique SNPs in the gyrB, PPE55, Rv2042c, leuS, mmpL6, and mmpS6 genes were used to determine their effectiveness as genetic markers for the identification of M. orygis. To the best of our knowledge, five of these SNPs, viz., gyrB 277 (A→G), gyrB 1478 (T→C), leuS 1064 (A→T), mmpL6 486 (T→C), and mmpS6 334 (C→G), are reported for the first time in this study. Our results also revealed several SNPs specific to other species within MTBC. The phylogenetic analysis shows that the studied genomes were genetically diverse and clustered with M. orygis sequences of human and animal origin reported from different geographic locations. Therefore, the present study provides a new insight into the high-confidence identification of M. orygis from MTBC species based on WGS data, which can be useful for reference and diagnostic laboratories.

Variation in Care of Well-Appearing Hypothermic Young Infants: A Multisite Study.

BACKGROUND AND OBJECTIVES: Numerous decision tools have emerged to guide management of febrile infants, but limited data exist to guide the care of young infants presenting with hypothermia. We evaluated the variation in care for well-appearing hypothermic young infants in the hospital and/or emergency department setting between participating sites. METHODS: This is a retrospective cohort study of well-appearing infants ≤90 days old across 9 academic medical centers from September 1, 2016 to May 5, 2021. Infants were identified via billing codes for hypothermia or an initial temperature ≤36.0°C with manual chart review performed. Primary outcomes included assessment of variation in diagnostic evaluation, disposition, empirical antimicrobial therapy, and length of stay. RESULTS: Of 14 278 infants originally identified, 739 met inclusion criteria. Significant interhospital variation occurred across all primary outcomes. Across sites, a full serious bacterial illness evaluation was done in 12% to 76% of hypothermic infants. Empirical antibiotics were administered 20% to 87% of the time. Performance of herpes simplex viral testing ranged from 7% to 84%, and acyclovir was empirically started 8% to 82% of the time. Hospital admission rates ranged from 45% to 100% of patients. CONCLUSIONS: Considerable variation across multiple aspects of care exists for well-appearing young infants presenting with hypothermia. An improved understanding of hypothermic young infants and their risk of infection can lead to the development of clinical decision tools to guide appropriate evaluation and management.

Patient satisfaction with access, affordability and quality of diabetes care at Mohalla Clinics in Delhi, India.

Introduction: Mohalla Clinics have been set up to provide curative care for minor ailments free of cost within walking distance in the urban slums, thus making primary care more accessible and affordable. Studies evaluating patient satisfaction with treatment of chronic conditions, such as diabetes, in these clinics are lacking. Methods: A survey of 400 type 2 diabetes patients was conducted, split equally between Mohalla clinics (MC) and Private clinics (PC) in Delhi. Responses were analyzed using STATA17, applying appropriate statistical tests for the data type (Chi-square test, Mann-Whitney U test, Wilcoxon signed rank test, or two-sample t test). Results: Satisfaction level was high in both groups with no significant difference between mean satisfaction scores of MC patients and PC patients (Mean 3.79 vs. 3.85 respectively, p = 0.4). However, MC patients reported a significant improvement in their satisfaction score after switching to MC (Mean 3.79 vs. 3.3 for the previous facility, p < 0.05). Physician interaction with the patients was the most important factor in influencing the satisfaction score. Proximity to the clinic was the second most important factor for MC patients but was not as important for PC patients. Surprisingly, treatment success was considered an important factor for satisfaction level by < 10% MC and < 20% PC patients only, pointing to the need for patient education across both the groups. None of the MC patients mentioned free treatment as a contributory factor to high satisfaction, perhaps because most shifted from a government setup to MC. PC patients had more frequent follow-up visits and blood glucose monitoring, and longer consultation duration compared to MC patients, which were offset by access factors, thus not causing much difference to the satisfaction score between the two groups. Conclusion: Mohalla clinics are making diabetes treatment accessible and affordable for the marginalized population of Delhi, despite not being designed or fully equipped to care for chronic diseases such as diabetes that require multi-specialty care to monitor and manage multiple co-morbidities and long-term complications. Positive perception of physician interaction and convenient location of the clinics are the two major contributors to the high satisfaction patients expressed with diabetes care at these clinics.

Prevalence of Invasive Bacterial Infection in Hypothermic Young Infants: A Multisite Study.

OBJECTIVE: To determine the prevalence of bacteremia and meningitis (invasive bacterial infection [IBI]) in hypothermic young infants, and also to determine the prevalence of serious bacterial infections (SBI) and neonatal herpes simplex virus and to identify characteristics associated with IBI. STUDY DESIGN: We conducted a retrospective cohort study of infants ≤90 days of age who presented to 1 of 9 hospitals with historical or documented hypothermia (temperature ≤36.0°C) from September 1, 2017, to May 5, 2021. Infants were identified by billing codes or electronic medical record search of hypothermic temperatures. All charts were manually reviewed. Infants with hypothermia during birth hospitalization, and febrile infants were excluded. IBI was defined as positive blood culture and/or cerebrospinal fluid culture treated as a pathogenic organism, whereas SBI also included urinary tract infection. We used multivariable mixed-effects logistic regression to identify associations between exposure variables and IBI. RESULTS: Overall, 1098 young infants met the inclusion criteria. IBI prevalence was 2.1% (95% CI, 1.3-2.9) (bacteremia 1.8%; bacterial meningitis 0.5%). SBI prevalence was 4.4% (95% CI, 3.2-5.6), and neonatal herpes simplex virus prevalence was 1.3% (95% CI, 0.6-1.9). Significant associations were found between IBI and repeated temperature instability (OR, 4.9; 95% CI, 1.3-18.1), white blood cell count abnormalities (OR, 4.8; 95% CI, 1.8-13.1), and thrombocytopenia (OR, 5.0; 95% CI, 1.4-17.0). CONCLUSIONS: IBI prevalence in hypothermic young infants is 2.1%. Further understanding of characteristics associated with IBI can guide the development decision tools for management of hypothermic young infants.

Premeal almond load decreases postprandial glycaemia, adiposity and reversed prediabetes to normoglycemia: A randomized controlled trial.

BACKGROUND: Asian Indians show rapid conversion from prediabetes to type 2 diabetes (T2D). Novel dietary strategies are needed to arrest this progression, by targeting postprandial hyperglycaemia (PPHG). DESIGN: We conducted a free-living randomized controlled open-label parallel arm study to evaluate the effect of a premeal load of almonds (20 g) 30 min before major meals on anthropometric, glycaemic, and metabolic parameters over 3 months. Sixty-six participants with prediabetes in the age range of 18-60 yrs were recruited. The study was registered at clinicaltrials.gov (registration no. NCT04769726). RESULTS: Thirty participants in each arm completed the study. As per 'intention-to-treat' analysis, overall additional mean reductions were statistically significant for body weight, BMI, waist circumference (WC), subscapular and suprailiac skinfolds, and improved handgrip strength (Kg) (p < 0·001 for all) in the treatment arm vs. the control arm (after multiple adjustments). In the blood parameters, the additional mean reduction in the treatment arm vs. control arm was statistically significant for fasting and post-75 g oral glucose-load blood glucose, postprandial insulin, HOMA-IR, HbA1c, proinsulin, total cholesterol, and very low-density lipoprotein cholesterol (p < 0·001 for all). Most importantly, we observed a reversal to normoglycemic state (fasting blood glucose and 2 h post-OGTT glucose levels) in 23.3% (7 out of 30) of participants in the treatment arm which is comparable to that seen with Acarbose treatment (25%). CONCLUSION: Incorporation of 20 g of almonds, 30 min before each major meal leads to significant improvement in body weight, WC, glycemia (particularly PPHG), and insulin resistance and shows potential for reversal of prediabetes to normal glucose regulation over 3 months.

Beneficial effects of premeal almond load on glucose profile on oral glucose tolerance and continuous glucose monitoring: randomized crossover trials in Asian Indians with prediabetes.

BACKGROUND: Rapid conversion from prediabetes to diabetes and frequent postprandial hyperglycemia (PPHG) is seen in Asian Indians. These should be the target of dietary strategies. OBJECTIVES: We hypothesized that dietary intervention of preloading major meals with almonds in participants with prediabetes will decrease overall glycemia and PPHG. DESIGN: The study included two phases: (1) an oral glucose tolerance test (OGTT)-based crossover randomized control study, the effect of a single premeal almond load (20 g) given before OGTT was evaluated (n = 60, 30 each period). (2) The continuous glucose monitoring system (CGMS)-based study for 3 days including premeal almond load before three major meals was a free-living, open-labeled, crossover randomized control trial, where control and premeal almond load diets were compared for glycaemic control (n = 60, 30 in each period). The study was registered at clinicaltrials.gov (registration no. NCT04769726). RESULTS: In the OGTT-based study phase, the overall AUC for blood glucose, serum insulin, C-peptide, and plasma glucagon post-75 g oral glucose load was significantly lower for treatment vs. control diet (p < 0.001). Specifically, with the former diet, PPHG was significantly lower (18.05% in AUC on OGTT, 24.8% at 1-h, 28.9% at 2-h post OGTT, and 10.07% during CGMS). The CGMS data showed that premeal almond load significantly improved 24-glucose variability; SD of mean glucose concentration and mean of daily differences. Daily glycaemic control improved significantly as per the following: mean 24-h blood glucose concentration (M), time spent above 7.8 mmol/L of blood glucose, together with the corresponding AUC values. Premeal almond load significantly decreased following: overall hyperglycemia (glucose AUC), PPHG, peak 24-h glycaemia, and minimum glucose level during night. CONCLUSION: Incorporation of 20 g of almonds, 30 min before each major meal led to a significant decrease in PPHG (as revealed in OGTT-based study phase) and also improved insulin, C-peptide, glucagon levels, and improved glucose variability and glycemic parameters on CGMS in participants with prediabetes. CLINICAL TRIAL REGISTRY: The study was registered at clinicaltrials.gov (registration no. NCT04769726).

Challenges for Lower-Middle-Income Countries in Achieving Universal Healthcare: An Indian Perspective.

Universal health coverage (UHC) by 2030 is a commitment of the global community adopted as Sustainable Development Goal 3.8. UHC, as defined by WHO, means all people have access to quality health services, when and where they need them, and without financial hardship. However, low-income and lower-middle-income countries, faced with competing priorities, find themselves struggling to muster enough resources to steer towards this goal at the desired pace. India is the largest lower-middle-income country, accounting for almost 18% of the world's population. How it performs in moving towards this goal will have a significant impact on achieving UHC at a global level. India has witnessed noteworthy improvement in several health indicators and the UHC service coverage index in recent decades, but the progress on improving service capacity and access has been rather slow given the enormity of its population, scarcity of funds, grossly inadequate public infrastructure, shortage of trained workforce, disparate needs of various regions of the country, lack of healthcare system integration, changing disease demography, and slack regulatory framework. The recent push through National Health Mission aims to address some of these challenges; however, a fragmented health delivery structure ossified over decades has been slow to keep up with the requirements of the country's massive and diverse population. This paper discusses the inherent characteristics and key challenges faced by the healthcare delivery system of India in achieving UHC.

Synergistic effect of sequential solvent treatment on the structural and low temperature charge transport of PEDOT:PSS films.

A detailed study on the low temperature charge transport and magnetoresistance (MR) measurements of spin-coated and solvent treated poly(3,4-ethylene dioxythiophene):poly(styrene sulfonate) (PEDOT:PSS) films has been presented. The samples were prepared by sequential treatment of PEDOT:PSS film using dimethyl sulfoxide and sulphuric acid. X-ray diffraction and Raman spectroscopy elucidate the effect of solvent treatment on the structural modifications of the film, which correlates to the enhanced conductivity values. The nature of solvent and the sequence of successive treatments using different solvents has been effectively utilized to tune the conductivity of the film over a wide range. Further, the low temperature (300-2 K) charge transport study indicates that the resistance of the film rises sharply below 50 K, which implies the samples lie in the category of disordered materials. A linear fit of lnRvs.T-1/4for all the samples indicates that the temperature dependent resistance of solvent treated PEDOT:PSS films follow 3D variable range hopping model. The observed large positive MR behavior of the films has been discussed in terms of the wave function shrinkage of the charge carriers. The MR follows ∼B2and ∼B1/3dependence at low and high magnetic field regimes, respectively. The sequential treatment of PEDOT:PSS films shown in the present study is an effective method to enhance the electrical conductivity significantly.

Compilation of 10 Years of MIRU-VNTR Data: Canadian National Tuberculosis Laboratory's Experience.

Tuberculosis is a significant cause of morbidity worldwide and is a priority at the provincial and federal levels in Canada. It is known that tuberculosis transmission networks are complex and span many years as well as different jurisdictions and countries. MIRU-VNTR is a universal tuberculosis genotyping method that utilizes a 24-loci pattern and it has shown promise in identifying inter and intrajurisdictional clusters within Canada. MIRU-VNTR data collected over 10 years from the National Reference Centre for Mycobacteriology (NRCM) were analyzed in this study. Some clusters were unique to a single province/territory, while others spanned multiple provinces and/or territories in Canada. The use of a universal laboratory test can enhance contact tracing, provide geographical information on circulating genotypes, and hence, aid in tuberculosis investigation by public health. The housing of all data on one platform, technical ease of the method, easy exchange of data between jurisdictions, and strong collaboration with laboratories and surveillance units at the provincial and federal levels have the potential to identify possible outbreaks in real time.

A Simple Assay for the Detection of Late-Stage Apoptosis Features in Saccharomyces cerevisiae.

Unicellular eukaryotic organisms such as yeast and protozoa serve as useful models for studying the impact of chemicals on cell physiology, cellular growth, and genome duplication. The yeast Saccharomyces cerevisiae has been widely used to assess apoptosis induced by chemicals due to its genetic tractability, ease of evaluation, and readily available impact assessment tools. Apoptosis in S. cerevisiae is characterized by many features, including increased cell death, loss of membrane integrity, release of caspases, chromatin condensation, and nuclear fragmentation, which are similar to the ones observed in mammalian cells. Current methods of apoptosis assessment typically require specialized equipment and reagents, which limits wide adoption. Here, we describe a rapid, inexpensive, and easy-to-perform assay in yeast for the analysis of late-stage apoptotic features in cells treated with a chemical. We describe a protocol for assessing loss of cell survival and changes in the nucleus. We demonstrate the approach by using acetic acid and hydrogen peroxide as test chemicals. This assay for the study of late-stage apoptotic features in S. cerevisiae can be performed reliably and rapidly by any laboratory with basic equipment and may be extended for studying apoptosis in similar single-cell organisms after treatment with toxicological agents. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Culture of Saccharomyces cerevisiae, treatment with acetic acid or hydrogen peroxide, and semi-quantitative growth assay Basic Protocol 2: DAPI staining and fluorescence microscopy for the assessment of change in nucleus-to-cytoplasm ratio and nuclear integrity.

An unusual location of solitary fibrous tumor in heart-A case report and review of literature.

BACKGROUND: Solitary fibrous tumor (SFT) are rare spindle cell tumors originating from the mesenchymal cells mostly from the visceral pleura. SFT was first described as a distinct entity in 1931 by Klemperer et al. Until now, we have limited data regarding the manifestation and behavior of extra pleural forms such as cardiac SFT. Here we present a case of SFT involving the pericardium where the diagnosis was made by imaging followed by biopsy findings. We also review the literature of SFT involving the heart and the management approaches. CASE PRESENTATION: An 81-year-old male presented with progressive dyspnea. Computed tomography (CT) of the chest showed a 6.2 × 5.3 cm soft tissue mass in the anterior mediastinum. Further imaging with CT angiogram showed a stalk-like connection to the pericardium. A biopsy of the mass showed spindle cells positive for BCL-2, CD34, and STAT 6, indicative of a solitary fibrous tumor. A surveillance approach was adopted for the patient. CONCLUSION: Primary pericardial tumors are exceedingly rare, with a prevalence rate of 0.001%-0.007%. Diagnosing a SFT requires a positive CD34 and BCL-2 marker. The current recommendation is resection of localized disease which has been documented to be curative in cases of benign disease however our patient was put on surveillance.

Deletion of autophagy related, ATG1 and F-box motif encoding YDR131C, together, lead to synthetic growth defects and flocculation behavior in Saccharomyces cerevisiae.

Ubiquitin proteasome system (UPS) and autophagy both pathways are involved in clearing the nonessential cellular components and also crosstalk during cellular response to normal and stress conditions. The F-box motif proteins constitute the SCF-E3 ligase complex of the UPS pathway in Saccharomyces cerevisiae and are involved in the substrate recruitment for ubiquitination. The ATG1 encoded Atg1p, a conserved serine-threonine kinase is crucial for the autophagy process. Here in this study, we report that loss of F-box motif encoding YDR131C and ATG1 together results in growth defects, floc formation, sensitivity to hydroxyurea, methyl methanesulfonate, and hydrogen peroxide. Both the genes also interact with the flocculation-related genes (FLO) and associate with gene ontology terms "ubiquitin-protein transferase activity" and "cellular catabolic process." Based on in silico analysis and experimental evidence we conclude that YDR131C and ATG1 function in parallel pathways to regulate the growth, flocculation, and stress response.

Deletion of Slc6a14 reduces cancer growth and metastatic spread and improves survival in KPC mouse model of spontaneous pancreatic cancer.

Pancreatic ductal adenocarcinoma (PDAC) is lethal. There is a dire need for better therapeutic targets. Cancer cells have increased demand for sugars, amino acids, and lipids and therefore up-regulate various nutrient transporters to meet this demand. In PDAC, SLC6A14 (an amino acid transporter (AAT)) is up-regulated, affecting overall patient survival. Previously we have shown using in vitro cell culture models and in vivo xenograft mouse models that pharmacological inhibition of SLC6A14 with α-methyl-l-tryptophan (α-MLT) attenuates PDAC growth. Mechanistically, blockade of SLC6A14-mediated amino acid transport with α-MLT leads to amino acid deprivation, eventually inhibiting mTORC1 signaling pathway, in tumor cells. Here, we report on the effect of Slc6a14 deletion on various parameters of PDAC in KPC mice, a model for spontaneous PDAC. Pancreatic tumors in KPC mice show evidence of Slc6a14 up-regulation. Deletion of Slc6a14 in this mouse attenuates PDAC growth, decreases the metastatic spread of the tumor, reduces ascites fluid accumulation, and improves overall survival. At the molecular level, we show lower proliferation index and reduced desmoplastic reaction following Slc6a14 deletion. Furthermore, we find that deletion of Slc6a14 does not lead to compensatory up-regulation in any of the other amino transporters. In fact, some of the AATs are actually down-regulated in response to Slc6a14 deletion, most likely related to altered mTORC1 signaling. Taken together, these results underscore the positive role SLC6A14 plays in PDAC growth and metastasis. Therefore, SLC6A14 is a viable drug target for the treatment of PDAC and also for any other cancer that overexpresses this transporter.