Normal Biopsies Research Articles

Role of needle biopsy in early diagnosis of lung lesions in tertiary care hospital

Lung cancer is one of the most common cancer worldwide and the leading cause of cancer‑related death. Needle biopsies of lung have been used for diagnosis of lung lesions. They are done CT guided or thoracoscopic guided procedures in our institution.The aim is to study role of needle biopsy in early diagnosis of lung lesions and incidence of lung lesions in needle biopsies in our hospital with respect to age and sex.A retrospective type of study. The histopathological records of our Hospital were reviewed for needle biopsies of lung lesions between January 2019 to December 2019 (One year). 35 cases noted during this period.There were a total of 35 cases during the study period of one year. The specimens received were linear bits and average size was 0.5cm in length. Non-neoplastic lesions were eight (22.85 %) and neoplasms contributed to 15 cases (42.85%) along with inadequate biopsies (20 %) and normal lung biopsies (14.28 %).Maximum number of cases were reported in the age group of 51-70 years (28 cases) with male preponderance. In all age groups, malignant lesions were more common in which, Adenocarcinoma was predominant.To conclude, in our study malignant neoplasms predominated over non neoplastic lesions. The most common histological type encountered in this study was an Adenocarcinoma.

Novel insights into the molecular mechanisms underlying risk of colorectal cancer from smoking and red/processed meat carcinogens by modeling exposure in normal colon organoids.

Tobacco smoke and red/processed meats are well-known risk factors for colorectal cancer (CRC). Most research has focused on studies of normal colon biopsies in epidemiologic studies or treatment of CRC cell lines in vitro. These studies are often constrained by challenges with accuracy of self-report data or, in the case of CRC cell lines, small sample sizes and lack of relationship to normal tissue at risk. In an attempt to address some of these limitations, we performed a 24-hour treatment of a representative carcinogens cocktail in 37 independent organoid lines derived from normal colon biopsies. Machine learning algorithms were applied to bulk RNA-sequencing and revealed cellular composition changes in colon organoids. We identified 738 differentially expressed genes in response to carcinogens exposure. Network analysis identified significantly different modules of co-expression, that included genes related to MSI-H tumor biology, and genes previously implicated in CRC through genome-wide association studies. Our study helps to better define the molecular effects of representative carcinogens from smoking and red/processed meat in normal colon epithelial cells and in the etiology of the MSI-H subtype of CRC, and suggests an overlap between molecular mechanisms involved in inherited and environmental CRC risk.

Prevalence Estimates of Predicted Pathogenic COL4A3-COL4A5 Variants in a Population Sequencing Database and Their Implications for Alport Syndrome.

The reported prevalence of Alport syndrome varies from one in 5000 to one in 53,000 individuals. This study estimated the frequencies of predicted pathogenic COL4A3-COL4A5 variants in sequencing databases of populations without known kidney disease. Predicted pathogenic variants were identified using filtering steps based on the ACMG/AMP criteria, which considered collagen IV α3-α5 position 1 Gly to be critical domains. The population frequencies of predicted pathogenic COL4A3-COL4A5 variants were then determined per mean number of sequenced alleles. Population frequencies for compound heterozygous and digenic combinations were calculated from the results for heterozygous variants. COL4A3-COL4A5 variants resulting in position 1 Gly substitutions were confirmed to be associated with hematuria (for each, P<0.001). Predicted pathogenic COL4A5 variants were found in at least one in 2320 individuals. p.(Gly624Asp) represented nearly half (16 of 33, 48%) of the variants in Europeans. Most COL4A5 variants (54 of 59, 92%) had a biochemical feature that potentially mitigated the clinical effect. The predicted pathogenic heterozygous COL4A3 and COL4A4 variants affected one in 106 of the population, consistent with the finding of thin basement membrane nephropathy in normal donor kidney biopsy specimens. Predicted pathogenic compound heterozygous variants occurred in one in 88,866 individuals, and digenic variants in at least one in 44,793. The population frequencies for Alport syndrome are suggested by the frequencies of predicted pathogenic COL4A3-COL4A5 variants, but must be adjusted for the disease penetrance of individual variants and for the likelihood of already diagnosed disease and non-Gly substitutions. Disease penetrance may depend on other genetic and environmental factors.

A Diagnostic Method for Gastric Cancer Using Two-Photon Microscopy With Enzyme-Selective Fluorescent Probes: A Pilot Study.

BackgroundEndoscopy is the most important tool for gastric cancer diagnosis. However, it relies on naked-eye evaluation by endoscopists, and the histopathologic confirmation is time-consuming. We aimed to visualize and measure the activity of various enzymes through two-photon microscopy (TPM) using fluorescent probes and assess its diagnostic potential in gastric cancer.Methodsβ-Galactosidase (β-gal), carboxylesterase (CES), and human NAD(P)H: quinone oxidoreductase (hNQO1) enzyme activities in the normal mucosa, ulcer, adenoma, and gastric cancer biopsy samples were measured using two-photon enzyme probes. The fluorescence emission ratio at long and short wavelengths (Ch2/Ch1) for each probe was comparatively analyzed. Approximately 8,000 – 9,000 sectional images in each group were obtained by measuring the Ch2/Ch1 ratio according to the tissue depth. Each probe was cross-validated by measuring enzymatic activity from a solution containing lysed tissue.ResultsTotal of 76 subjects were enrolled in this pilot study (normal 21, ulcer 18, adenoma 17, and cancer 20 patients, respectively). There were significant differences in the mean ratio values of β-gal (0.656 ± 0.142 vs. 1.127 ± 0.109, P < 0.001) and CES (0.876 ± 0.049 vs. 0.579 ± 0.089, P < 0.001) between the normal and cancer, respectively. The mean ratio value of cancer tissues was different compared to ulcer and adenoma (P < 0.001). The hNQO1 activity showed no significant difference between cancer and other conditions. Normal mucosa and cancer were visually and quantitatively distinguished through β-gal and CES analyses using TPM images, and enzymatic activity according to depth, was determined using sectional TPM ratiometric images. The results obtained from lysis buffer-treated tissue were consistent with TPM results.ConclusionsTPM imaging using ratiometric fluorescent probes enabled the discrimination of gastric cancer from normal, ulcer, and adenoma. This novel method can help in a visual differentiation and provide quantitative depth profiling in gastric cancer diagnosis.

A body map of somatic mutagenesis in morphologically normal human tissues.

Somatic mutations that accumulate in normal tissues are associated with ageing and disease1,2. Here we performed a comprehensive genomic analysis of 1,737 morphologically normal tissue biopsies of 9 organs from 5 donors. We found that somatic mutation accumulations and clonal expansions were widespread, although to variable extents, in morphologically normal human tissues. Somatic copy number alterations were rarely detected, except for in tissues from the oesophagus and cardia. Endogenous mutational processes with the SBS1 and SBS5 mutational signatures are ubiquitous among normal tissues, although they exhibit different relative activities. Exogenous mutational processes operate in multiple tissues from the same donor. We reconstructed the spatial somatic clonal architecture with sub-millimetre resolution. In the oesophagus and cardia, macroscopic somatic clones that expanded to hundreds of micrometres were frequently seen, whereas in tissues such as the colon, rectum and duodenum, somatic clones were microscopic in size and evolved independently, possibly restricted by local tissue microstructures. Our study depicts a body map of somatic mutations and clonal expansions from the same individual.

Evaluation of the Impact of Intratumoral Heterogeneity of Esophageal Cancer on Pathological Diagnosis and P16 Methylation and the Representativity of Endoscopic Biopsy.

BackgroundP16 methylation is expected to be potential diagnostic and therapeutic targets for esophageal cancer (EC). The intratumoral heterogeneity (ITH) of EC has been mentioned but has not been quantitatively measured yet. We aimed to clarify the impact of ITH on pathological diagnosis and P16 methylation, and the concordance between endoscopic biopsy and the corresponding surgically resected tissue.MethodsWe designed a systematic sampling method (SSM) compared with a general sampling method (GSM) to obtain EC tumor tissue, tumor biopsy, and normal squamous epithelium biopsy. MethyLight assay was utilized to test P16 methylation. All specimens obtained by the SSM were pathologically diagnosed.ResultsA total of 81 cases were collected by the GSM, and 91.4% and 8.6% of them were esophageal squamous cell carcinomas (ESCCs) and esophageal adenocarcinomas (EADs), respectively. Nine SSM cases were 100.0% ESCCs. The positive rates of P16 methylation of the GSM tumor and normal tissues were 63.0% (51/81) and 32.1% (26/81), respectively. For SSM samples, tumor tissues were 100.0% (40/40) EC and 85.0% (34/40) P16 methylated; tumor biopsy was 64.4% (29/45) diagnosed of EC and 68.9% P16 methylated; the corresponding normal biopsies were 15.7% (8/51) dysplasia and 54.9% (28/51) P16 methylated. The concordance of pathological diagnosis and P16 methylation between tumor biopsy and the corresponding tumor tissue was 75.0% and 62.5%, respectively.ConclusionThe SSM we designed was efficient in measuring the ITH of EC. We found inadequate concordance between tumor biopsy and tissue in pathological diagnosis and P16 methylation.

Transcriptome-wide In Vitro Effects of Aspirin on Patient-derived Normal Colon Organoids.

Mechanisms underlying aspirin chemoprevention of colorectal cancer remain unclear. Prior studies have been limited because of the inability of preclinical models to recapitulate human normal colon epithelium or cellular heterogeneity present in mucosal biopsies. To overcome some of these obstacles, we performed in vitro aspirin treatment of colon organoids derived from normal mucosal biopsies to reveal transcriptional networks relevant to aspirin chemoprevention. Colon organoids derived from 38 healthy individuals undergoing endoscopy were treated with 50 μmol/L aspirin or vehicle control for 72 hours and subjected to bulk RNA sequencing. Paired regression analysis using DESeq2 identified differentially expressed genes (DEG) associated with aspirin treatment. Cellular composition was determined using CIBERSORTx. Aspirin treatment was associated with 1,154 significant (q < 0.10) DEGs prior to deconvolution. We provide replication of these findings in an independent population-based RNA-sequencing dataset of mucosal biopsies (BarcUVa-Seq), where a significant enrichment for overlap of DEGs was observed (P < 2.2E-16). Single-cell deconvolution revealed changes in cell composition, including a decrease in transit-amplifying cells following aspirin treatment (P = 0.01). Following deconvolution, DEGs included novel putative targets for aspirin such as TRABD2A (q = 0.055), a negative regulator of Wnt signaling. Weighted gene co-expression network analysis identified 12 significant modules, including two that contained hubs for EGFR and PTGES2, the latter being previously implicated in aspirin chemoprevention. In summary, aspirin treatment of patient-derived colon organoids using physiologically relevant doses resulted in transcriptome-wide changes that reveal altered cell composition and improved understanding of transcriptional pathways, providing novel insight into its chemopreventive properties. PREVENTION RELEVANCE: Numerous studies have highlighted a role for aspirin in colorectal cancer chemoprevention, though the mechanisms driving this association remain unclear. We addressed this by showing that aspirin treatment of normal colon organoids diminished the transit-amplifying cell population, inhibited prostaglandin synthesis, and dysregulated expression of novel genes implicated in colon tumorigenesis.

Potency of miR-144-3p in promoting abdominal aortic aneurysm progression in mice correlates with apoptosis of smooth muscle cells

Abdominal aortic aneurysm (AAA), a life-threatening disease, is commonly diagnosed among people with risk factors, including increasing age, male gender, and smoking. The apoptosis of smooth muscle cells (SMCs) has been reported to disrupt the vascular structural integrity, which causes AAA. Thus, we sought to characterize the potential role of microRNA (miR)-144-3p in SMC apoptosis, and to outline the molecular mechanisms involved in this pathway. We collected pathological abdominal aortic tissues and adjacent normal aortic biopsy specimens from 18 patients undergoing AAA repair surgery. The relationship between miR-144-3p expression and SMC proliferation was assessed by transfecting mimic/inhibitor of miR-144-3p in human aortic smooth muscle cells (HASMCs). Anti-growth effect of miR-144-3p and related genes was evaluated in a murine AAA model. Dual luciferase reporter gene assay was adopted to validate the targeting relationship between miR-144-3p and enhancer of zeste homolog 2 (EZH2), and the enrichment of EZH2 in the p21 promoter region was determined by chromatin immunoprecipitation assay. MiR-144-3p was highly expressed in AAA tissues. Enhanced miR-144-3p diminished SMC proliferation by binding to the EZH2 3′-untranslated region and thereby inhibiting EZH2 expression. In addition, EZH2 was highly enriched in the promoter region of p21, and knockdown of p21 expression could rescue the effect of miR-144-3p on SMC proliferation and apoptosis. miR-144-3p serves as a promoter for the apoptosis of SMCs, which contributes to the occurrence and progression of AAA. This observation will serve as the basis for further investigations into potential p21-based therapies for AAA treatment.

Repeat Biopsy to Assess Duodenal Healing in Children With Celiac Disease and Eosinophilic Gastrointestinal Disorders.

A retrospective chart review was performed of children with CD and EGID seen at the Children's Hospital of Philadelphia between 2003 and 2018. Data collected included duodenal biopsy pathology, celiac serology including tTG-IgA, and symptom reports. Duodenal healing was defined as normal villous architecture and no intraepithelial lymphocytes. These data were compared with tTG-IgA level. Data were analyzed with Fisher exact test and t test methods. Thirty-nine patients had normal IgA and diagnoses of both CD and EGID. At second biopsy, 44% (17/39) of patients showed no histologic evidence of active CD and 36% (14/39) of patients had negative tTG-IgA values. Sixty percent (9/15) of patients with no evidence of CD on biopsy had abnormal tTG-IgA levels, and 57% (8/14) of patients with normal tTG-IgA levels had evidence of active disease on biopsy. The data show that an abnormal tTG-IgA drawn after initiation of a gluten-free diet is not correlated with duodenal mucosal injury in pediatric patients with CD and EGID. This suggests that serologic surveillance with tTG-IgA is not sufficient to monitor CD intestinal healing in this patient cohort. Persistent elevations of tTG-IgA in CD patients with normal duodenal biopsies should prompt investigation into other potential causes.

Calmodulin Antagonist W-7 Enhances Intermediate Conductance Ca2+- Sensitive Basolateral Potassium Channel (IKCa) Activity in Human Colonic Crypts

Intermediate conductance potassium (IKCa) channels are exquisitively Ca2+ sensitive, intracellular Ca2+ regulating channel activity by complexing with calmodulin (CaM), which is bound to the cytosolic carboxyl tail. Although CaM antagonists might be expected to decrease IKCa channel activity, the effect of W-7 in human T lymphocytes are conflicting. We therefore evaluated the effect of W-7 on basolateral IKCa channels in human colonic crypt cells. Intact crypts obtained from normal human colonic biopsies by Ca2+ chelation were used for patch clamp studies of basolateral IKCa channels in the cell-attached configuration. IKCa channel activity was studied when the bath Ca2+ concentration was changed from 1.2 mmol/L to 100 μmol/L and back to 1.2 mmol/L, as well as from 100 μmol/L to 1.2 mmol/L and back to 100 μmol/L, both in the absence and presence of 25 μmol/L W-7. Decreasing bath Ca2+ from 1.2 mmol/L to 100 μmol/L decreased IKCa channel activity reversibly in the absence of W-7, whereas there was a uniformly high level of channel activity at both bath Ca2+ concentrations in the presence of W-7. In separate experiments, increasing bath Ca2+ from 100 μmol/L to 1.2 mmol/L increased IKCa channel activity reversibly in the absence of W-7, whereas there was again a uniformly high level of channel activity at both bath Ca2+ concentrations in the presence of W-7. We, therefore, propose that W-7 has a specific stimulatory effect on basolateral IKCa channel activity, despite its ability to inhibit Ca2+/CaM-mediated, IKCa channel-dependent Cl− secretion in human colonic epithelial cells.Graphic

QS10: Morphological Changes Of Skin Related To Acellular Dermal Matrix Incorporation In Tissue Expansion

Purpose:Acellular dermal matrix (ADM) is used to create an inferolateral sling nearly three-quarters of the time in breast reconstruction and has proven a valuable alternative to total submuscular coverage of the implant. Previous work has demonstrated decreased inflammation and fibrosis of the pocket lining the ADM sling; however there has been minimal investigation into the role of ADM in skin growth and regeneration. The present study evaluates morphologic and molecular changes mediated by use of ADM in tissue expansion.Methods:Two tissue expanders, one wrapped in ADM, were placed subcutaneously on the back of Yucatan minipigs. All expanders were inflated with two weekly fills of 60cc of normal saline and skin biopsies were harvested after two weeks of expansion from each condition: control, tissue expansion (TE), and tissue expansion with ADM (TE+ADM). Three biopsies per condition were embedded in paraffin or OCT medium and stained with Russell Movat Pentachrome and Immunofluorescence of CD31, respectively. Collagen in the papillary dermis of pentachrome-stained images were analyzed using an ImageJ plug-in, Fibril Tool, that applies circular statistics to estimate average fibril orientation as the direction angle from -90 to 90 with respect to the x-axis. One-way ANOVA evaluated seventy-two measurements per condition and post-hoc analysis with Tukey’s HSD test identified significant comparisons between the groups. Number of fluorescent cells expressing CD31 (a marker of endothelial cells) were counted on 12 photographs per condition. P-values ≤ .05 were considered significant. Total deformation was calculated using a computational model and isogeometric analysis.Results:The mean fibril orientation of TE and TE+ADM underwent -85% change (P < .001) and -15% change (P = .65), respectively, compared to control. Three times more CD31+ cells were observed in TE+ADM compared to control (P < .001), but no significant changes were detected in TE alone. Histogram of total deformation revealed more even distribution of forces in TE+ADM compared to TE and control.Conclusions:The use of ADM in a porcine tissue expansion model appears to mitigate disarray of the collagen network in adjacent tissue, thereby creating a more extensive, yet even, distribution of stretched skin. This observation, combined with the finding of increased angiogenesis, suggests it is the incorporation of ADM that confers these protective benefits. Future studies will evaluate whether the protective effects of ADM can serve to improve TE in compromised tissue beds, as seen in patients undergoing TE concurrent with perioperative radiation therapy.

Interstitial amyloidosis in sporadic inclusion body myositis.

Intracellular congophilic inclusions within muscle fibers, although nonspecific, are one of the pathological hallmarks of sporadic inclusion body myositis (sIBM). Extracellular amyloid deposits in muscle, on the other hand, are the canonical findings of amyloid myopathies, which occur with or without systemic amyloidosis. We reviewed the muscle biopsy database (1998-2020) to identify sIBM patients with extracellular amyloid deposits. Clinical and laboratory data were reviewed. We identified five sIBM patients (three clinicopathologically defined and two clinically defined) with extracellular amyloid deposits in muscle. Mean age at diagnosis was 74.8y (range, 68-84 y). All patients had a typical sIBM pattern of weakness without associated sensory or autonomic symptoms. None had electrophysiological evidence of peripheral neuropathy. Only one patient had a monoclonal gammopathy (immunoglobulin M-lambda, IgM-λ) with normal bone marrow biopsy. This patient with monoclonal gammopathy and three other patients underwent abdominal fat pad aspirate and were negative for amyloid. Cardiac evaluation was unrevealing in the four patients tested. Three patients without monoclonal gammopathy had normal transthyretin gene sequencing and inconclusive mass spectrometry-based analysis. The patient with monoclonal gammopathy died of pneumosepsis 5y after diagnosis and autopsy revealed multi-organ transthyretin amyloidosis. Detection of extracellular amyloid deposition in muscle should trigger an aggressive search for systemic amyloidosis independently from other associated myopathological abnormalities. Amyloid subtyping is crucial for early therapy and mortality prevention. An isolated monoclonal gammopathy should not halt a search for non-hematological causes of systemic amyloidosis.

On the feasibility of deep learning applications using raw mass spectrometry data.

SummaryIn recent years, SWATH-MS has become the proteomic method of choice for data-independent–acquisition, as it enables high proteome coverage, accuracy and reproducibility. However, data analysis is convoluted and requires prior information and expert curation. Furthermore, as quantification is limited to a small set of peptides, potentially important biological information may be discarded. Here we demonstrate that deep learning can be used to learn discriminative features directly from raw MS data, eliminating hence the need of elaborate data processing pipelines. Using transfer learning to overcome sample sparsity, we exploit a collection of publicly available deep learning models already trained for the task of natural image classification. These models are used to produce feature vectors from each mass spectrometry (MS) raw image, which are later used as input for a classifier trained to distinguish tumor from normal prostate biopsies. Although the deep learning models were originally trained for a completely different classification task and no additional fine-tuning is performed on them, we achieve a highly remarkable classification performance of 0.876 AUC. We investigate different types of image preprocessing and encoding. We also investigate whether the inclusion of the secondary MS2 spectra improves the classification performance. Throughout all tested models, we use standard protein expression vectors as gold standards. Even with our naïve implementation, our results suggest that the application of deep learning and transfer learning techniques might pave the way to the broader usage of raw mass spectrometry data in real-time diagnosis.Availability and implementationThe open source code used to generate the results from MS images is available on GitHub: https://ibm.biz/mstransc. The raw MS data underlying this article cannot be shared publicly for the privacy of individuals that participated in the study. Processed data including the MS images, their encodings, classification labels and results can be accessed at the following link: https://ibm.box.com/v/mstc-supplementary.Supplementary information Supplementary data are available at Bioinformatics online.

Methylation profile of colon cancer genes in colorectal precursor lesions and tumor tissue: perspectives for screening

Aims Epigenetic alterations of genes involved in colorectal carcinogenesis are likely to be informative biomarkers for early detection. We assessed the methylation profile of a panel of seven colon cancer-related genes comparing normal colon, colorectal cancer (CRC) precursor lesions and cancer tissues from a Brazilian cohort. Methods The cohort comprised 114 CRC patients, including 40 matched normal tissue, 47 patients with adenomas, 33 with serrated polyps and 8 with normal colonic biopsy. DNA methylation status of SEPT9, ALX4, NDRG4, BMP3, APC, p16 and MLH1 was determined by pyrosequencing and correlated with clinicopathological features. Sensitivity, specificity, positive predictive value and negative predictive value were calculated for all genes using cancer endpoint. Results The most frequently methylated genes in cancer and in precancer lesions were SEPT9, ALX4, NDRG4, and BMP3, ranging from 55.3 to 95% of the samples. Overall, the frequency of methylation of these four genes in normal colonic tissue was significantly lower as compared to cancer or precursor lesions both in adenoma–carcinoma (p < .001 and p < .050) and serrated (sessile-serrated lesion) (p < .001 and p < .050) pathways. Additionally, sensitivity for the cancer endpoint ranged from 65.6 to 91.8%, and specificity from 17.9 to 62.9% for SEPT9, ALX4, NDRG4, and BMP3 genes. Moreover, the comethylation of ≥4 genes was higher in sessile-serrated lesion (87.5%) and conventional adenomas (78.7%) than in hyperplastic polyps (43.7%) (p = .025) and was significantly associated with proximal cancers (p = .042). Conclusions Our study suggests the DNA methylation can constitute potential biomarkers in CRC screening of Brazilian population.

A humanized monoclonal antibody against the endothelial chemokine CCL21 for the diagnosis and treatment of inflammatory bowel disease.

Chemokines are small proteins that promote leukocyte migration during development, infection, and inflammation. We and others isolated the unique chemokine CCL21, a potent chemo-attractant for naïve T-cells, naïve B-cells, and immature dendritic cells. CCL21 has a 37 amino acid carboxy terminal extension that is distinct from the rest of the chemokine family, which is thought to anchor it to venule endothelium where the amino terminus can interact with its cognate receptor, CCR7. We and others have reported that venule endothelium expressing CCL21 plays a crucial role in attracting naïve immune cells to sites of antigen presentation. In this study we generated a series of monoclonal antibodies to the amino terminus of CCL21 in an attempt to generate an antibody that blocked the interaction of CCL21 with its receptor CCR7. We found one humanized clone that blocked naïve T-cell migration towards CCL21, while memory effector T-cells were less affected. Using this monoclonal antibody, we also demonstrated that CCL21 is expressed in the mucosal venule endothelium of the large majority of inflammatory bowel diseases (IBD), including Crohn’s disease, ulcerative colitis, and also in celiac disease. This expression correlated with active IBD in 5 of 6 cases, whereas none of 6 normal bowel biopsies had CCL21 expression. This study raises the possibility that this monoclonal antibody could be used to diagnose initial or recurrent of IBD. Significantly, this antibody could also be used for therapeutic intervention in IBD by selectively interfering with recruitment of naïve immune effector cells to sites of antigen presentation, without harming overall memory immunity.

Managing Abnormal Uterine Bleeding: A Comparative Study of Modified Thermal Balloon Ablation and Hysterectomy

Objective: To compare the outcome of modified thermal balloon ablation and hysterectomy for management of heavy menstrual bleeding in terms of frequency of patient satisfaction. Methods: All women aged between 35 to 45 years with heavy menstrual bleeding and who have completed their family were selected for the study. All 267 patients fulfilling inclusion criteria were recruited with uterine length less than 12cm, without any intrauterine pathology and normal endometrial biopsy. Results: Group A Patients underwent hysterectomy and patient's satisfaction rate was 91.3% (126/138). Group B patients underwent modified thermal balloon ablation and patient's satisfaction rate was 82 .6% (114 / 138). Conclusion: In the management of heavy menstrual bleeding, results of hysterectomy and modified thermal balloon ablation are comparable to each other in terms of achievement of patient's satisfaction especially when we have to choose between definitive and conservative surgical options.

Biventricular Strain Imaging with Cardiac MRI in Genotyped and Histology Validated Amyloid Cardiomyopathy.

Cardiac amyloidosis (CA) is a common and potentially fatal infiltrative cardiomyopathy. Contrast-enhanced cardiac MRI (CMR) is used as a diagnostic tool. However, utility of CMR for the comprehensive analysis of biventricular strains and strain rates is not reported as extensively as echocardiography. In addition, RV strain analysis using CMR has not been described previously.Objectives:We sought to study the global and regional indices of biventricular strain and strain rates in endomyocardial biopsy (EMB)-proven, genotyped cases of CA.Methods:A database of 80 EMBs was curated from 2012 to 2019 based on histology. A total of 19 EMBs positive for CA were subjected to further tissue-characterization with histology, and compared with four normal biopsy specimens. Samples were genotyped for ATTR- or AL-subtypes. Five patients, with both echocardiography and contrast-enhanced CMR performed 72-h apart, were subjected to comprehensive analysis of biventricular strain and strain-rates.Results:Histology confirmed that the selected samples were indeed positive for cardiac amyloid. Echocardiography showed reduced global and regional left-ventricular (LV) longitudinal strain indices. CMR with tissue-characterization of LV showed global reductions in circumferential, radial and longitudinal strains and strain-rates, following a general trend with the echocardiographic findings. The basal right-ventricular (RV) segments had reduced circumferential strains with no changes in longitudinal strain.Conclusions:In addition to providing a clinical diagnosis of CA based on contrast clearance-dynamics, CMR can be a potent tool for accurate functional assessment of global and regional changes in strain and strain-rates involving both LV and RV. Further studies are warranted to validate and curate the strain imaging capacity of CMR in CA.

Association Between a Common, Benign Genotype and Unnecessary Bone Marrow Biopsies Among African American Patients

Up to two-thirds of African American individuals carry the benign rs2814778-CC genotype that lowers total white blood cell (WBC) count. To examine whether the rs2814778-CC genotype is associated with an increased likelihood of receiving a bone marrow biopsy (BMB) for an isolated low WBC count. This retrospective genetic association study assessed African American patients younger than 90 years who underwent a BMB at Vanderbilt University Medical Center, Mount Sinai Health System, or Children's Hospital of Philadelphia from January 1, 1998, to December 31, 2020. The rs2814778-CC genotype. The proportion of individuals with the CC genotype who underwent BMB for an isolated low WBC count and had a normal biopsy result compared with the proportion of individuals with the CC genotype who underwent BMB for other indications and had a normal biopsy result. Among 399 individuals who underwent a BMB (mean [SD] age, 41.8 [22.5] years, 234 [59%] female), 277 (69%) had the CC genotype. A total of 35 patients (9%) had clinical histories of isolated low WBC counts, and 364 (91%) had other histories. Of those with a clinical history of isolated low WBC count, 34 of 35 (97%) had the CC genotype vs 243 of 364 (67%) of those without a low WBC count history. Among those with the CC genotype, 33 of 34 (97%) had normal results for biopsies performed for isolated low WBC counts compared with 134 of 243 individuals (55%) with biopsies performed for other histories (P < .001). In this genetic association study, among patients of African American race who had a BMB with a clinical history of isolated low WBC counts, the rs2814778-CC genotype was highly prevalent, and 97% of these BMBs identified no hematologic abnormality. Accounting for the rs2814778-CC genotype in clinical decision-making could avoid unnecessary BMB procedures.

Analysis of Mitochondrial Metabolic Profile As a Predictor of Clinical Aggressiveness of Acute Myeloid Leukemia

Introduction:Acute myeloid leukemia (AML) is a clonal hematopoietic malignancy characterized by dysregulated differentiation, unrestricted growth, and inhibition of apoptosis of immature myeloid progenitors. A feature in AML is increased metabolic demand despite genetic heterogeneity. Mitochondria regulate ATP generation, metabolism, apoptosis and are central to proliferation and differentiation in normal and cancer cells. The Translocase of the Outer Mitochondrial Membrane Member 20 (TOMM20), a member of the outer membrane translocator complex, plays a critical role in import of precursor proteins like nuclear encoded oxidative phosphorylation (OXPHOS) subunits from cytosol to mitochondria, required for ATP generation. TOMM20 expression and uptake of the mitotracker (a fluorescent dye taken up by mitochondria with intact membrane potential), correlate with functional mitochondrial mass and OXPHOS activity. We aim to characterize the metabolic profile of AML, using TOMM20 as a surrogate marker of mitochondrial OXPHOS, and to assess in a subgroup of subjects, differences in mitotracker uptake between AML and non-cancer cells.Methods:This is a retrospective evaluation of 67 patients with AML treated at Thomas Jefferson University between 2005 and 2015. The patients were divided into 3 cohorts with (a) relapsed disease post-hematopoietic stem cell transplantation (HSCT) [n=20], (b) remission post-HSCT [n=27] and (c) refractory disease post induction therapy [n=20].The study hypotheses were: (1) Mitochondrial metabolism measured by TOMM20 expression is increased in AML compared to normal bone marrow biopsy specimens and there is higher mitotracker uptake in AML cells than non-cancer bone marrow cells (2) Biologic behavior of AML can be predicted by TOMM20 expression with patients who relapse having higher expression than those that maintain a remission and (3) At relapse, there will be an increase in TOMM20 expression from initial diagnosis with the patient serving as his/her own control. Immunostaining of TOMM20 in bone marrow specimens was graded by two pathologists (blinded to clinical characteristics) as 1+: faint fine granular staining in areas of maximum staining, 2+: coarse granular staining with distinctly visible granules in areas of maximum staining and 3+: coarse coalesced granular staining in areas of maximum staining. Overall survival in relation to TOMM20 expression was measured.The association of TOMM20 expression with unpaired categorical data was performed using chi-square or Fisher's exact test and paired data using the non-parametric Wilcoxon rank-sum test. Kaplan-Meier test was used to evaluate survival rates and compared using the log-rank test. Mitotracker deep red mean fluorescent intensity in CD34+ and CD34 bone marrow cells in AML were compared using a paired t-test.Results:The median age at AML diagnosis was 58 (range: 20-78) years and 63% were female. Per the Revised Disease Risk Index, patients fell into low (13%), intermediate (64%) and high (23%) risk groups. In the control bone marrow specimens analyzed (n=6), only scattered myeloid precursors showed 1+ TOMM20 expression compared to AML blasts that showed uniform 1+ to 3+ staining [figure 1]. Mitotracker uptake was 7.7 fold higher (p<0.05) in AML cells than in non-cancer bone marrow cells in 5 subjects with AML whose cancer cells expressed CD34.There was no significant difference in TOMM20 expression between the 3 patient groups at diagnosis (p=0.26). In the patient group relapsing post HSCT, there was a statistically significant increase in median TOMM20 expression compared to baseline (Z=-3.2, p=0.001). Patients with high TOMM20 expression (3+) had the lowest overall survival compared to those with 1+ or 2+ expression, although not statistically significant (Log rank p value=0.217) [figure 2].Discussion:We demonstrate that AML blasts have increased TOMM20 expression and mitotracker uptake, markers of mitochondrial metabolism, compared to normal myeloid precursors. We did not observe a significant difference in TOMM20 expression at diagnosis between patient cohorts. In AML patients who relapse post HSCT, there is an increase in functional mitochondrial mass as demonstrated by increased expression of TOMM20. Mitochondrial OXPHOS inhibitors are promising oncotherapeutic agents warranting prospective evaluation in treatment strategies for relapsed and/or refractory AML. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

The Relationship Between Hyperplastic Gastric Polyps And Helicobacter Pylori

Objective&#x0D; Helicobacter pylori (H. pylori) is a gram-negative bacterium. It is urease positive, mobile, microaerophilic, and spiral-shaped, and it settles in the stomach and the proximal duodenum. It is associated with chronic gastritis, stomach ulcers, duodenal ulcers, stomach cancer, and MALT lymphoma. It can also cause premalignant lesions, such as metaplasia and gastric atrophy. Although several studies are investigating the correlation between H. pylori and hyperplastic polyps (HPs) of the stomach, uncertainty in this regard continues. &#x0D; Methods&#x0D; Data from 5378 patients who underwent esophagogastroduodenoscopy at the Department of Gastroenterology, Gülhane Training and Research Hospital, &#x0D; University Of Health Sciences between October 2016 and March 2019 were retrospectively analyzed, and patients with gastric polyps were evaluated for polyp pathologies and normal mucosa biopsies.&#x0D; Results&#x0D; Sixty-one stomach polyps were detected in 49 patients after the retrospective analysis of the data between October 2016 and March 2019. Of the polyps found, 36 were hyperplastic (73%). There was no statistically significant difference between HPs and the presence of H. pylori, age, gender, anatomical location of polyps in the stomach, size, number, presence of metaplasia, or atrophy. The mean age was statistically lower in H. pylori-positive cases.&#x0D; Conclusions&#x0D; No correlation was observed between HPs and the presence of H. pylori. There is a need for more extensive, multi-centered, and prospective studies evaluating the relationship between HPs and H. pylori.