Unveiling Preeclampsia Prognosis: Uterine Artery Doppler Indices in Low-Risk Pregnancies.

Background Anticipating preeclampsia's onset is pivotal in mitigating adverse maternal and perinatal outcomes. This study aims to prognosticate preeclampsia within low-risk pregnancies by evaluating uterine artery Doppler indices within the 14-28 week gestation. Methodology An observational cohort comprising 360 low-risk pregnancies (14-28 weeks gestation) underwent serial uterine artery Doppler assessments at 14-20 and 20-28 weeks. Follow-up was extended to delivery to detect preeclampsia incidence. Results Among 360 participants, 56 (15.5%) developed preeclampsia. Sensitivity values for resistance index (RI), pulsatility index (PI), and bilateral notching were 17.6%, 56.25%, and 71%, respectively, during 14-20 weeks. Similarly, during 20-28 weeks, sensitivities for RI, PI, and bilateral notching were 16.6%, 36.8%, and 55.5%, respectively, with specificity exceeding 90%. Notch depth index (NDI) >0.14 emerged as a better predictor of preeclampsia between both intervals (area under the curve = 0.686 and 0.646). Conclusions Bilateral notching during 14-20 weeks and NDI >0.14 within 14-20 and 20-28 weeks indicate preeclampsia susceptibility in low-risk pregnancies. Conversely, uterine artery Doppler indices at 14-28 weeks effectively rule out preeclampsia development, exhibiting a specificity of >90%.

Analysis of Different Routes of Hysterectomy Based on a Prospective Algorithm and Their Complications in a Tertiary Care Institute.

Introduction: Hysterectomy is the most common gynaecological operation worldwide. The objective of the study is to analyze the various routes of hysterectomy and its complications when the decision of route is based on using a prospective algorithm tree. Methodology. It is an observational study to analyze the route of hysterectomy based on using a prospective algorithm. The decision tree is based on pelvic pathology, uterine size, vaginal access, pelvic adhesion, competency of the surgeon, choice of the patient, and complication of different routes of hysterectomy. Data were collected from preoperative, intraoperative, and postoperative records. Demographic factors, indications, routes of hysterectomy, and complications were recorded and analyzed by using SPSS software version 22. Observation. Among the malignant or suspected malignant pathology groups, TAH was performed in 89 cases and TLH was performed in 3 cases. Among the benign disease groups, VH was performed in 137(38.2%) cases, TAH was performed in 118(32.9%) cases, and TLH was performed in 104 (28.9%) cases. Operative time and a number of blood transfusions were significantly less with VH (p value < 0.0001 and 0.004) compared to abdominal and total laparoscopic hysterectomy. Postoperative complication such as fever was more with abdominal hysterectomy (p-value<0.00001) compared to VH and TLH. Vaginal discharge was more with VH and TLH compared to TAH (p value -0.004) and wound infection was more in the abdominal route (p value 0.001). Conclusion: The abdominal route was the route of choice for surgery in malignancy or suspected malignant pathology. In benign pathology, VH was the most common and preferable route of surgery. Complications were found to be minimal with vaginal hysterectomy.

Teaching and Learning Medical Professionalism: an Input from Experienced Faculty and Young Graduates in a Tertiary Care Institute.

Introduction: Medical professionalism is of paramount importance especially in today's day and age. This study gives an insight on the preferred methods of teaching and learning professionalism among the young MBBS graduates and experienced faculty in a medical teaching institute. Material and methods: This was an observational study conducted in the Department of Obstetrics and Gynaecology from January 2019 to January 2020. It included 60 interns and 60 faculty members, who were interviewed for nine different methods of teaching and learning professionalism: (a) professional role model; (b) early clinical exposure; (c) recruiting faculty who had compressive training in medical education before joining the medical institute; (d) teaching and assessing communication skills to each student; (e) conducting seminar, didactic lecture and small group discussion; (f) reflective practice; (g) mentorship; (h) faculty development programme; (i) hidden curriculum. Each participant's response was analyzed using Wilcoxon rank-sum test on SPSS software version 22. Results:Interns preferred early clinical exposure, recruiting faculty with prior comprehensive training in medical education and reflective practice as preferred methods, while faculty members preferred teaching and assessing communication skills for every student, early clinical exposure and mentorship. Conclusion:Early clinical exposure, teaching and assessing communication skills, mentorship and reflective practice are the preferred methods of teaching and learning medical professionalism.

A rare case of misoprostol hypersensitivity.

Medical abortion using mifepristone-misoprostol regimen has been considered to be a safe and effective method for pregnancy termination. Misoprostol is a frequently used well tolerated drug with mild and transient side effects. Considering the safety profile mifepristone-misoprostol regimen has also been advocated by many for home-based medical abortion. However, we report herein a rare case of hypersensitivity reaction to sublingual misoprostol administered for first trimester medical abortion, where timely diagnosis and prompt intervention prevented life-threatening airway obstruction. The possibility of such rare event should be kept in mind and included in patient counselling and information especially for those who opt for outpatient abortion care so that they can seek medical help at the earliest.

Amniotic Fluid Embolism After First-Trimester Abortion.

Amniotic fluid embolism (AFE) may be a rare event in pregnancy, especially after a first-trimester medical termination of pregnancy (MTP). A 35-year-old G3P2L2 came to our outpatient department at six weeks of pregnancy for medical termination of pregnancy and bilateral tubal ligation. After around one hour of surgery, she developed respiratory distress with abdominal distension, hypotension, tachycardia and tachypnoea. On laparotomy, we found ascitic fluid, bowels with petechia, and oozing all over the wounds. Finally, within 24 hours of surgery, she expired. Strong clinical suspicion of AFE should prompt a multidisciplinary team including anaesthesia, respiratory therapy, critical care, and maternal-foetal medicine to be involved in the ongoing care of women with AFE.

An Unusual Case of Retained Foreign Body in the Vagina With Vaginal Fibrosis.

Vaginal foreign bodies can cause long haul, foul-smelling vaginal discharge and vaginal bleeding and are typically found in female youngsters while looking into vaginitis and urinary tract diseases. There are many causes for vaginal discharges. Among them, vaginal foreign bodies are uncommon but not a very rare presentation. We had a case of a 49-year-old female, a widow, with para 2 living 2 and a history of menopause since three years; she was referred from a district hospital with a diagnosis of carcinoma of the cervix and was later found to have a foreign body, which was removed surgically through the vagina. A foreign body in the vagina is usually seen in children than in adults. Foreign bodies are inserted vaginally for treatment purposes, contraception, induced abortion, and sexual stimulation in adults. Here, we report a case of retained vaginal foreign body with vaginal fibrosis.

Maternal mortality and predictors of adverse outcome in patients with heart disease in pregnancy.

Introduction: Heart disease in pregnancy possesses a great haemodynamic challenge and is a known risk for increased maternal morbidity and mortality. The functional status of the patient is one of the most significant parameters which can impact the feto-maternal outcome. Many predictors have been studied and compiled in various scoring systems time and again. The most updated and validated is the modified WHO classification, according to which the presence of pulmonary artery hypertension (PAH) and severe ventricular dysfunction (ejection fraction <30%) mandates the patient to be under class IV, which along with another important risk factor, i.e., New York heart association (NYHA) class, is revaluated under the present study. The objective of this study is to examine three of the most important predictors of adverse outcomes, i.e., functional status (NYHA class), PAH, and left ventricular ejection fraction (LVEF) in patients with heart disease in pregnancy. Methods: It's a prospective study from January 2016 to August 2017 wherein pregnant patients with heart disease were divided on the basis of NYHA class, PAH, and LVEF, and the feto-maternal outcome was recorded and evaluated in terms of maternal mortality, fetal demise, the occurrence of major cardiac complication, and risk of preterm delivery. Results: A total of three out of 29 (10.34%) maternal deaths were attributed to a cardiac cause. 5.45% of patients with heart disease had maternal mortality, which is in contrast to the 1.12% maternal mortality rate in general at our centre. Three out of 17 (17.64%) patients in NYHA classes 3 and 4 ended in maternal deaths, while there were no mortalities in classes 1 and 2. Intrauterine fetal demise (23.52%), risk of preterm delivery (relative risk = 0.4688; 95% CI: 0.2320 to 0.9470) was significantly higher in patients belonging to NYHA classes 3 and 4 as compared to those in classes 1 and 2. All of the ten (100%) patients who developed cardiac complications belonged to classes 3 and 4. The percentage of abortions (20.00%), intra uterine fetal demise (IUFD) (40.00%), and cardiac complication (80%) in patients with LVEF <44% were significantly more than in patients with better ejection fraction. Pulmonary artery systolic pressure (PASP) ≥ is associated with higher maternal mortality, a greater number of abortions and IUFD (22.62%), cardiac complication (22.72%), and increased risk of preterm birth (0.5769; 95% CI: 0.2801 to 1.188), but these associations are not found to be significant. Conclusion: NYHA class was found to be a very strong predictor followed by left ventricular ejection fraction for poor outcome. Maternal mortality in asymptomatic patients or patients with mild symptoms (NYHA classes 1 and 2) is comparable to that found in the general population. However, pulmonary artery systolic pressure is not found to be significantly associated with worse outcomes in our study.

A study to assess the knowledge and awareness among young doctors about emergency contraception.

BACKGROUND: Emergency contraception (EC) is the contraception on demand which can prevent millions of unintended pregnancies. The knowledge and awareness of young doctors towards EC who may be the first contact physician of the society has not been well studied. This study aims to assess the knowledge and awareness of young doctors in a teaching institute in northeast India. METHODOLOGY: This study was carried out among 200 young doctors and included 100 interns and 100 postgraduate trainees (PGT) and senior resident doctors (SRD) from January 2020 to March 2020 to compare their knowledge and awareness about EC. A predesigned self-administered 22 items questionnaire was used to collect data. OBSERVATION: In our study, majority of the doctors in both groups were aware of levonorgestrel 1.5 mg tablet as EC (93% and 95%) and more interns than PG SRD were aware of its easy availability (86%, 35%, P value < 0.0001), government supply (77%, 30%, P value < 0.0001), and that copper intrauterine contraceptive device (IUCD) can be used as EC up to 120 h (89%, 60%, P value < 0.0001). Most doctors were unaware of ulipristal acetate. Most PGT SRDs believe that EC promotes irresponsible behavior, sexually transmitted diseases, and promiscuity but most intern did not agree to it (P value < 0.0001 for each). More than 65% doctors in both groups were aware of the mechanism of action of EC. PGT SRD were more aware of the effectiveness of EC (62%, 80%, P value 0.0078). More interns were aware that EC affects the next period (53%, 25%, P value < 0.0001). CONCLUSION: Interns were more aware about contraception than PGT and SRD, especially about government supply of EC, about IUCD, and behavioral aspect like promoting irresponsible behavior, sexually transmitted disease, and promiscuity.

Targeted metagenomics using next generation sequencing in laboratory diagnosis of culture negative endophthalmitis.

To study the feasibility of 16S rRNA metagenomics using next generation sequencing (NGS) along with broad range PCR assay for 762 bp region of 16S rRNA gene with Sanger's sequencing, in microbial diagnosis of culture negative endophthalmitis. Vitreous fluid from 16 culture negative and one culture positive endophthalmitis patients, admitted to a tertiary care hospital were processed for targeted metagenomics. NGS of 7 variable regions of 16S rRNA gene was done using Ion Torrent Personal Genome Machine (PGM). Sequence data were analyzed using Ion Reporter software using QIIME and BLSATN tools and Greengenes and NCBI-Genbank databases. Bacterial genome sequences were detected in 15 culture negative and culture positive vitreous specimens. The sequence reads varied between 25,245-540,916 with read length between 142bp-228bp and coverage depth was 41.0X and 81.2X. Operational taxonomic unit (OTUs) of multiple bacterial genera and species were detected in 13 culture negative vitreous specimens and OTUs of a single bacterial species were detected in 2 culture negative and 1 culture positive specimens; one negative specimen had no bacterial DNA. Maximum numbers of OTUs detected by NGS for a bacterial species from any vitreous specimen was the one which was detected and identified by Sanger's sequencing in broad range PCR. All the bacteria were belonging to clinically relevant species. Broad range PCR with sequencing failed to identify bacteria from 5 of the 16 (31.25%) culture negative vitreous specimens. Metagenomics could detect and identify bacterial pathogens in 15 of the 16 culture negative vitreous specimen's up to species level. With rapidly decreasing cost, metagenomics has a potential to be used widely in endophthalmitis diagnosis, in which culture negativity is usually high.

Impact of physical activity on mitochondrial enzymes, muscle stem cell and anti-oxidant protein Sestrins in Sarcopenic mice.

INTRODUCTION: Sarcopenia is the loss of skeletal muscle mass and function. It is a major health issue in old age due to lack of understanding of the origin and molecular mechanism. Altered dietary pattern, sedentary lifestyle and physical inactivity have shown adverse effect of skeletal muscle function. Sedentary behaviour and low protein intake have been well associated with sarcopenia. Here, we aim to develop Sarcopenia mimicking murine model to observe the physiological and biochemical changes with physical activity intervention. We also intended to find the association of muscle stem cells and stress induced protein Sestrins in the developed sarcopenic model. METHODS: Male C57BL/6 mice were categorized into 4 groups: young-control (Y-Cntrl), aged-matched control (A-Cntrl), Sarcopenic-model (SAR-model) and Sarcopenic intervention group (SAR-INT) with physical exercises. SAR-model group was kept in a retrofitted confined cage for sedentary lifestyle and was fed with protein-restricted diet. Phenotypic assessment for body mass, grip strength and functional endurance was analysed to confirm the sarcopenic state. Mitochondrial enzymatic assessment, muscle stem cell (MuSCs) proliferation potential and protein quantification of Sestrins expression were performed by enzyme histochemistry, flow cytometry and surface plasmon resonance (SPR), respectively. SAR-model group was given 10 weeks physical activity intervention to assess the physiological and biochemical changes. RESULTS: Simultaneous implementation of physical inactivity by sedentary confinement and protein restricted diet led the animals to exhibit the features of sarcopenia. SAR-model group showed a decline of 8.6% (p < 0.0001) in the body weight assessment, 32% decline (p < 0.0001) in the grip strength, 28% increase in time elapsed (p < 0.0001) indicating decline in functional performance. Mitochondrial enzymes (ATPase, NADH-TR and SDH/COX) assessment exhibited low expression in SAR-model group. Ki67 positive muscle stem cell declines around 50% in the model group. SPR quantification of Sestrin 2 showed a decline of 14% which significantly improved to 28% upon physical activity intervention (p = 0.0025) in SAR-INT group. CONCLUSION: It can be summarized that the mouse model generated in the present study mimics the feature of human Sarcopenia. Physical activity intervention may improve the sarcopenic status via modulation of Sestrin 2 which can serve as potential molecule for therapeutic implication.

Maternal and Perinatal Outcomes in Hypertensive Disorders of Pregnancy and Factors Influencing It: A Prospective Hospital-Based Study in Northeast India.

Introduction Hypertensive disorders frequently complicate pregnancy and contribute substantially to maternal and perinatal morbidity and mortality. Identification of risk factors for hypertensive disorders of pregnancy (HDP) can help determine the particular patient group which requires appropriate intervention. Methods This prospective cross-sectional hospital-based study conducted from January 2016 to January 2019 included all pregnant women beyond 20 weeks of gestation complicated by HDP. The objectives were to determine the incidence of HDP and associated maternal and perinatal mortality and morbidity rates along with factors influencing it. Data collected were entered in Microsoft Excel (Microsoft Corporation, Redmond, WA) and analyzed with the Statistical Package for the Social Sciences (SPSS) software version 21 (IBM Corp. Armonk, NY). Results In our study, out of 5460 deliveries, 402 (7.4%) cases had HDP, 27.6% had gestational hypertension, 27.6% had mild preeclampsia, 33.6% had severe preeclampsia, and 11.2% had eclampsia. Fifty-four (13.4%) cases required admission in the intensive care unit and 12 (2.9%) ended in maternal deaths. The cause of maternal mortality was cerebral hemorrhage in eight (66.6%) cases and pulmonary edema in four (33.3%) cases. All maternal deaths occurred in women with severe preeclampsia and eclampsia and eclampsia was significantly higher. Maternal deaths were more when systolic blood pressure (SBP) was ≥ 160mmHg, diastolic blood pressure (DBP) was ≥ 110mmHg, significantly more with 3+ proteinuria, but no association was found with age, parity, booking status, socio-economic status, gestational age, or mode of delivery. All mothers with HDP received treatment with antihypertensives. There were 60 (14.9%) cases of perinatal mortality. Perinatal deaths were more in unbooked cases and preterm HDP, significantly more with SBP ≥160 mmHg, DBP ≥110 mmHg and ≥2+proteinuria, but no association was found with parity or mode of delivery. Besides mortality, there was a significant burden of maternal and perinatal morbidity, which was more in women with severe preeclampsia and eclampsia. Conclusion Routine antenatal screening for HDP in all pregnant women with appropriate and timely interventions in women at risk may help reduce HDP-related maternal and perinatal morbidity and mortality.

Uterine Didelphys with Transverse Vaginal Septum - A Complex Rare Müllerian Anomaly.

During the development of the female genital tract, any insult to the normal development process results in a set of intriguing abnormalities known as Müllerian duct abnormalities. The uterine didelphys is the second least common type of anomaly among these, which may commonly be associated with a longitudinal vaginal septum (lateral fusion defect). However uterine didelphys along with a transverse vaginal septum (lateral fusion plus resorption defect) is a very rare finding and to the best of our knowledge, thecase that we hereby report is the second one in literature. A 16-year-old unmarried girl presented with primary amenorrhoea and cyclical pain for 18months.On clinical examination and imaging, a case of uterine didelphys and transverse vaginal septum was found. Her urinary tract was normalon USG and MRI evaluation. Excision of the septum was done by abdomino-vaginal approach. The patient was discharged well. We conclude that a patient presenting with primary amenorrhea especially with cyclical dysmenorrhea with a transverse vaginal septum on examination should be thoroughly investigated for associated upper genital tract abnormalities as the treatment strategy and prognosis is largely dependent on the correct classification of the anomaly.

Evaluation of 16S rRNA broad range PCR assay for microbial detection in serum specimens in sepsis patients.

BACKGROUND: Early and accurate laboratory diagnosis and appropriate management of infection improves the survival rate in sepsis. In this study we evaluated broad range 16S rRNA and 16 S-23 S intergenic spacer region (ISR) PCR assays followed by nucleotide sequencing directly from patients' serum and automated blood culture for laboratory diagnosis in admitted sepsis patients. METHODS: A broad range 16S rRNA PCR and 16 S-23 S ISR PCR assay followed by nucleotide sequencing was used directly from patients' serum in hospital admitted patients in 62 sepsis and 16 suspected blood stream infection (sBSI) patients. Automated blood culture was also used in the same patients. Nucleotide sequences were analyzed against NCBI Genbank database and organisms were identified using CLSI MM18A guidelines. RESULTS: Bacterial culture were positive in 10/62 (16.12%) sepsis and 3/16 (18.75%) suspected BSI patients along with 3 detected fungi (2 in sepsis and 1 in suspected BSI group). PCR assay was positive in 36/62 (58.06%) sepsis and 6/16 (37.5%) suspected BSI patients respectively. All but 2 bacteria (both from culture negative patients) detected by PCR assay could be identified from nucleotide sequencing. Survival in sepsis patients was 77%. PCR assay could detect bacteria in 9/14 (64.28%) of sepsis patients with death. CONCLUSION: Broad range PCR assay was far superior for early diagnosis of infection. The bacteria which could not be detected by culture and were not commonly reported from this centre, were detected by the broad range PCR assays. Detection of these rare bacteria/fungi had significant clinical correlation with patient's underlying clinical conditions, immune status and prognosis. The tests could provide definitive diagnosis of infection in >58% of sepsis patients, which helped in patient management and better survival.

Utility of broad-range 16S rRNA PCR assay versus conventional methods for laboratory diagnosis of bacterial endophthalmitis in a tertiary care hospital.

BACKGROUND: Endophthalmitis, a sight-threatening intraocular infection, can be of postsurgical, post-traumatic or endogenous origin. Laboratory diagnosis-based appropriate therapy can be vision-saving. Conventional culture-based laboratory diagnosis takes time and lacks sensitivity. In this study a broad-range PCR assay was assessed against conventional and automated culture methods in vitreous specimens for accurate microbiological diagnosis. AIMS: To use broad-range PCR assay targeting 16S ribosomal RNA (rRNA) region of bacteria and to assess its performance vis-à-vis conventional and automated culture methods in the laboratory diagnosis of endophthalmitis. METHODS: Vitreous specimens from 195 patients with clinically diagnosed endophthalmitis were processed for classical and automated culture methods, antimicrobial sensitivity and broad-range PCR assay targeting 762 bp region of 16S rRNA followed by nucleotide sequencing by Sanger's method. Causative agents were identified from the nucleotide sequences analysed against the GenBank database, and organisms were identified using the Clinical and Laboratory Standards Institute (CLSI) MM18A guidelines. RESULTS: Bacteria could be detected from 127 (65.13%) of the 195 vitreous specimens by broad-range PCR assay; bacterial isolation was possible from 17 (8.7%) and 60 (30.76%) of these specimens by conventional and automated culture methods, respectively (p<0.0001). PCR assay could detect two uncultured bacterium, and in five cases the bacterial identity could not be determined from NCBI database matching. CONCLUSION: Broad-range PCR assay could provide definitive microbial diagnosis within 24 hours in significantly more patients (p<0.0001). Some rare organisms could be detected, useful in treatment modalities. Automated culture was significantly more sensitive than conventional culture.

Analyzing Thyroid Dysfunction in the Climacteric.

INTRODUCTION: Perimenopause and menopause is an integral part of every woman's life constituting about 30% of the Indian female population. Hypothyroidism is a condition that can mimic menopausal symptoms very often. Therefore, routine screening of thyroid function in the climacteric and menopausal period to determine subclinical thyroid disease can be recommended. MATERIALS AND METHODS: With this background, we conducted a prospective study in our institute, North Eastern Indira Gandhi Regional Institute of Health and Medical Sciences, to evaluate the abnormal thyroid function tests in perimenopausal and postmenopausal women aged 40-80 years. RESULTS AND OBSERVATIONS: Subclinical hypothyroidism status increases from the age group of 40-70 years and hypothyroidism increases in slow frequency from 40 to 70 years after that there is plateauing. Interestingly, the frequency of hyperthyroidism remains same in all the age groups. CONCLUSION: Looking at the increased risks of thyroid disorders with age, screening of thyroid disorders can be started at the age of 40 years with estimation of serum levels of TSH. This gives us the opportunity to treat both subclinical and clinical hypothyroidism and hyperthyroidism women.

Diagnostic Hysterolaparoscopy for Evaluation of Infertility: Our Experience in a Tertiary Care Hospital.

OBJECTIVE: The objective of this study is to analyze the role of diagnostic hysterolaparoscopy (DHL) for evaluation of infertility in a tertiary care hospital. MATERIALS AND METHODS: This retrospective study was conducted from July 2014 to June 2016. RESULTS: Out of 151 patients, 58.28% and 41.72% had primary and secondary infertility, respectively. In primary infertility group 37.5% and in secondary infertility group 49.2% had abnormal findings. Most common finding was adnexal adhesions (pelvic inflammatory disease) and laparoscopic findings were more common than the hysteroscopic ones. CONCLUSION: DHL was helpful in finding some reversible causes of infertility such as adnexal adhesions, tubal blockade, and uterine synechiae, etc.

Hepatitis E Virus (HEV) egress: Role of BST2 (Tetherin) and interferon induced long non- coding RNA (lncRNA) BISPR.

BACKGROUND: The biology of Hepatitis E Virus (HEV), a common cause of epidemic and sporadic hepatitis, is still being explored. HEV exits liver through bile, a process which is essential for its natural transmission by feco-oral route. Though the process of this polarised HEV egress is not known in detail, HEV pORF3 and hepatocyte actin cytoskeleton have been shown to play a role. METHODS: Our transcriptome analysis in Hepatitis E virus (HEV) replicon transfected Huh7 cells at 24 and 72 hrs indicated that at 24hrs, both LncBISPR and BST2, expressed by a bidirectional promoter were highly upregulated whereas at 72 hrs, BST2 expression was comparatively reduced accompanied by normal levels of BISPR. These findings were confirmed by qPCR analysis. Co-localisation of BST2 and HEV pORF2 was confirmed in HEV transfected Huh7 by confocal microscopy. To investigate the role of BISPR/BST2 in HEV life cycle, particularly virus egress, we generated Huh7 cells with ~8kb deletion in BISPR gene using Crispr-Cas9 system. The deletion was confirmed by PCR screening, Sanger sequencing and Real time PCR. Virus egress in ΔBISPR Huh7 and Huh7 cells was compared by measuring HEV positive strand RNA copy numbers in cell lysates and culture supernatants at 24 and 72 hrs post HEV replicon transfection and further validated by western blot for HEV pORF2 capsid protein. RESULTS: ΔBISPR Huh7 cells showed ~8 fold increase in virus egress at 24 hrs compared to Huh7 cells. No significant difference in virus egress was observed at 72hrs. Immunohistochemistry in histologically normal liver and HEV associated acute liver failure revealed BST2 overexpression in HEV infected hepatocytes and a dominant canalicular BST2 distribution in normal liver in addition to the cytoplasmic localisation reported in literature. CONCLUSIONS: These findings lead us to believe that BISPR and BST2 may regulate egress of HEV virions into bile in vivo. This system may also be used to scale up virus production in vitro.

Identification of High-Risk Aberrant Crypt Foci and Mucin-Depleted Foci in the Human Colon With Study of Colon Cancer Stem Cell Markers.

BACKGROUND: During colonoscopic screening, only macroscopic lesions will be identified, and these are usually the result of multiple genetic abnormalities. Magnification endoscopic detection of aberrant crypt foci (ACF), long before they acquire complex genetic abnormalities, is promising. However, the features of high-risk ACF-like lesions need to be identified. MATERIALS AND METHODS: In the present cross-sectional study, grossly visible normal mucosal flaps were shaved from 152 colectomies, including 96 colorectal cancer (CRC) cases and 56 controls (22 control specimens with disease with malignant potential and 34 without malignant potential). Methylene and Alcian blue stains were performed directly on the unfixed mucosal flaps to identify ACF and mucin-depleted foci (MDF). Detailed topographic analyses, with immunohistochemical staining for ß-catenin and cancer stem cell (CSC) markers (CD44, CD24, and CD166) were performed. RESULTS: ACF, MDF, and ß-catenin-accumulated crypts were detected more in specimens with adjacent CRC. The left colon had ACF with a larger diameter and greater crypt multiplicity, density, and gyriform pit pattern and were considered the high-risk ACF group. MDF, more commonly associated with dysplasia, is also a marker of possible carcinogenesis. The CD44 CSC marker was significantly upregulated in ACF specimens compared with normal controls. Our 3-tier ACF-only pit pattern classification system showed better linearity with mucosal dysplasia than did the 6-tier Kudo classification. CONCLUSION: High-risk ACF, when detected during chromoendoscopic screening, should be followed up. CSCs might play an important role in pathogenesis. Larger studies and genotypic risk stratification for definite identification of high-risk ACF are needed.

Patients with mild enteropathy have apoptotic injury of enterocytes similar to that in advanced enteropathy in celiac disease.

BACKGROUND: Severity of villous atrophy in celiac disease (CeD) is the cumulative effect of enterocyte loss and cell regeneration. Gluten-free diet has been shown to benefit even in patients having a positive anti-tissue transglutaminase (tTG) antibody titre and mild enteropathy. AIM: We explored the balance between mucosal apoptotic enterocyte loss and cell regeneration in mild and advanced enteropathies. METHODS: Duodenal biopsies from patients with mild enteropathy (Marsh grade 0 and 1) (n=26), advanced enteropathy (Marsh grade ≥2) (n=41) and control biopsies (n=12) were subjected to immunohistochemical staining for end-apoptotic markers (M30, H2AX); markers of cell death (perforin, annexin V); and cell proliferation (Ki67). Composite H-scores based on the intensity and distribution of markers were compared. RESULTS: End-apoptotic markers and marker of cell death (perforin) were significantly up-regulated in both mild and advanced enteropathies, in comparison to controls; without any difference between mild and advanced enteropathies. Ki67 labelling index was significantly higher in crypts of mild enteropathy, in comparison to controls, suggesting maintained regenerative activity in the former. CONCLUSIONS: Even in patients with mild enteropathy, the rate of apoptosis is similar to those with advanced enteropathy. These findings suggest the necessity of reviewing the existing practice of not treating patients with mild enteropathy.