Regulatory Role of B Cells and Its Subsets in Hepatitis E Virus Infection.

Antibodies as well as memory B cells are the potential correlates of a protective immune response against hepatitis E virus (HEV) infection. Literature on the role of B regulatory cells (Bregs) in acute viral infections is limited. We have evaluated the role of IL-10 expressing Bregs in HEV infection. A total of 108 acute hepatitis E patients, 55 hepatitis E recovered individuals and 128 HEV naïve healthy controls were enrolled. The percentages of peripheral CD19+, immature CD19+CD24hiCD38hi, mature CD19+CD24intCD38int and memory CD19+CD24hiCD38- B cells were analyzed by flowcytometry. Intracellular cytokine staining for IL-10 and TGF-ß, HEV-rORF2p specific T cell response (IFN-γ expression) pre/post IL-10/IL-10R blocking and CD19+IL-10+ B cells-depletion based assays were carried out to assess the functionality of Bregs. The percentage of HEV-rORF2p specific immature B cell phenotype was significantly higher in acute hepatitis E patients compared to hepatitis E recovered individuals and controls. Significantly higher IL-10 expression on B and HEV-rORF2p stimulated immature B cells of acute hepatitis E patients compared to controls indicated that Bregs are functional and HEV-rORF2p specific. Enhanced IFN-γ expression on CD8+ T cells upon IL-10/IL-10R blocking and also post CD19+IL-10+ B cells depletion suggested that CD3+CD8+IFN-γ + T cells corroborate the regulatory potential of Bregs via IL-10 dependent mechanism. We have identified HEV specific functional, immature CD19+CD24hiCD38hi B cells having IL-10 mediated regulatory activities and a potential to modulate IFN-γ mediated T cell response in Hepatitis E. The prognostic/pathogenic role of Bregs in recovery from severe hepatitis E needs evaluation.

The role of imaging in pulmonary hypertension.

Pulmonary hypertension (PH) is a debilitating and potentially life threatening condition in which increased pressure in the pulmonary arteries may result from a variety of pathological processes. These can include disease primarily involving the pulmonary vasculature, but more commonly PH may result from left-sided heart disease, including valvular heart disease. Chronic thromboembolic pulmonary hypertension (CTEPH) is an important disease to identify because it may be amenable to surgical pulmonary artery endarterectomy or balloon pulmonary angioplasty. Parenchymal lung diseases are also widespread in the community. Any of these disease processes may result in adverse remodeling of the right ventricle and progressive right heart (RH) failure as a common final pathway. Because of the breadth of pathological processes which cause PH, multiple imaging modalities play vital roles in ensuring accurate diagnosis and classification, which will lead to application of the most appropriate therapy. Multimodality imaging may also provide important prognostic information and has a role in the assessment of response to therapies which ultimately dictate clinical outcomes. This review provides an overview of the wide variety of established imaging techniques currently in use, but also examines many of the novel imaging techniques which may be increasingly utilized in the future to guide comprehensive care of patients with PH.

An outbreak of hepatitis E in Yavatmal, India, 2019.

Hepatitis E, a public health concern in developing countries, frequently presents in epidemic, as well as in sporadic forms. This study investigated an outbreak of viral hepatitis at Yavatmal, Maharashtra, India in March 2019. Blood samples from 10 patients were received at Indian Council of Medical Research-National Institute of Virology, Pune to test for the presence of enterically transmitted hepatitis viruses. Subsequently, 49 suspected cases were screened for anti-hepatitis E virus (HEV)/hepatitis A virus (HAV) immunoglobulin M and immunoglobulin G (IgG) antibodies, alanine amino-transferase levels and HEV RNA. Water samples were screened for HEV and HAV RNA followed by phylogenetic analysis. Overall 32 of 49 (65.3%) suspected cases had recent acute HEV infection, while dual infection with HAV was noted in one case (2.04%). Forty-eight of 49 suspected cases were positive for anti-HAV IgG antibodies indicative of previously acquired immunity against HAV. Water samples had evidence of HEV contamination as detected by reverse transcription-polymerase chain reaction. Sequencing of HEV RNA from both patients (n = 2) and water samples (n = 5) indicated HEV genotype 1 to be the etiological agent of this outbreak. Serological and molecular evidence confirmed HEV as the etiology. Mixing of contaminated drain water with the domestic water supply may have triggered this outbreak. Subsequent changing of the defaulted water pipelines and its segregation from drain pipelines by the health authorities resulted in progressive decline of this outbreak. Despite the limitations, periodic surveillance of HEV exposure pattern and reporting of small outbreaks would supplement to the global disease burden data of hepatitis E.

Loss of DNA mismatch repair proteins in prostate cancer.

Recent studies have suggested an increased risk of prostate cancer in men with Lynch syndrome driven by germline mutations in mismatch repair (MMR) genes. However, the incidence and clinical implication of MMR deficiency in sporadic prostate cancers remain poorly understood. We immunohistochemically stained for MLH1, MSH2, MSH6, and PMS2 in a set of tissue microarray consisting of 220 radical prostatectomy specimens and evaluated the relationship between loss of their expression and available clinicopathological features. MLH1, MSH2, MSH6, and PMS2 were lost in 2 (0.9%), 6 (2.7%), 37 (16.8%), and 27 (12.3%) prostate cancers, respectively. Loss of at least 1 MMR protein was identified in 50 (22.7%) cases. There were no statistically significant associations between MMR deficiency and patient age, family history of prostate cancer, Gleason score, or pT/pN stage. Nonetheless, the levels of preoperative prostate-specific antigen (PSA) were significantly (P = .015) higher in patients with MMR deficiency (mean ±â€ŠSD: 9.12 ±â€Š9.01 ng/mL) than in those without abnormal MMR (5.76 ±â€Š3.17 ng/mL). There were 15 (6.8%) cases showing loss of at least 2 MMR proteins, which was not significantly associated with PSA level or tumor grade/stage. Additionally, 5 and 2 cases showed losses of at least 3 MMR proteins and all 4 proteins, respectively. Kaplan-Meier analysis revealed no significant associations between loss of MLH1 (P = .373), MSH2 (P = .348), MSH6 (P = .946), or PMS2 (P = .681), or at least 1 (P = .477), 2 (P = .486), or 3 (P = .352) MMR proteins and biochemical recurrence. Further analyses of the data on programmed death-ligand 1 (PD-L1) expression previously stained in the same set of tissue microarray demonstrated associations between loss of ≥2 MMR proteins and a higher rate of PD-L1 expression in cancer cells (17.2% vs 5.2%; P = .033) as well as between cases showing both loss of ≥1 MMR protein(s) and PD-L1 expression in tumor-infiltrating immune cells vs a higher risk of biochemical recurrence (P = .045). MMR protein loss was seen in a subset of prostate cancers. Interestingly, it was associated with significantly higher levels of PSA. Moreover, immunohistochemical detection of MMR proteins together with other proteins, such as PD-L1, might be helpful in predicting tumor recurrence following radical prostatectomy.

The impact of routine frozen section analysis during partial cystectomy for bladder cancer on surgical margin status and long-term oncologic outcome.

OBJECTIVES: The utility of frozen section analysis (FSA) during partial cystectomy has not been established. We assessed the impact of intraoperative FSA in partial cystectomy cases on surgical margin (SM) status and patient outcome. SUBJECTS AND METHODS: A retrospective review identified 76 consecutive patients who underwent partial cystectomy for bladder carcinoma with (n = 66; 87%) or without (n = 10; 13%) FSA for SMs at our institution from 2004 to 2018. FSA was correlated with the diagnosis of the frozen section control, the status of final SM, and the prognosis. RESULTS: Final SM was positive in 9 (12%) cystectomies, including 6 (9%) FSA vs. 3 (30%) non-FSA cases (P = 0.091). There were no significant differences in tumor size, histology, or tumor grade/stage between the 2 cohorts. FSAs were reported as positive (n = 7; 11%), atypical (n = 10; 15%), and negative (n = 49; 74%). All of the positive and negative FSA diagnoses were confirmed accurate on the frozen section controls, whereas atypical diagnoses were revised to benign (n = 4), atypical (n = 4), and carcinoma (n = 2) on the controls. Ten (77%) of 13 initial FSA-positive (6 of 7)/atypical (4 of 6; excluding benign diagnoses on the controls) cases achieved negative conversion by excision of additional tissue. Thus, final SM was positive in 1 (14%) FSA-positive case, 3 (30%) FSA-atypical cases (including one at the SM where FSA was not sampled), and 2 (4%) FSA-negative cases (at the SM where FSA was not sampled). Kaplan-Meier analysis and log-rank test revealed an association of performing FSA with the risk of disease progression (P = 0.021), but not intravesical recurrence (P = 0.434) or cancer-specific mortality (P = 0.560). Initial positive/atypical FSA, as an independent prognosticator, was associated with reduced progression-free (P = 0.002) and cancer-specific (P = 0.004) survival rates, compared with initial negative FSA. Positive SM was also associated with a larger tumor size (P < 0.05) and a higher risk of intravesical recurrence (P = 0.070) or disease progression (P = 0.096). CONCLUSIONS: Performing FSA during partial cystectomy may contribute to preventing positive SM and disease progression. Additionally, as seen in most of initial FSA-positive/atypical cases that achieved negative conversion, select patients may benefit from the routine FSA. Meanwhile, positive or atypical FSA was associated with significantly poorer prognosis.

Study of a hepatitis E virus outbreak involving drinking water and sewage contamination in Shimla, India, 2015-2016.

BACKGROUND: Hepatitis E, caused by hepatitis E virus (HEV), accounts for 50% of acute hepatitis cases in India. We report an outbreak of hepatitis E in Shimla, India, in 2015-2016. METHODS: ICMR-National Institute of Virology (NIV), Pune, received two batches of water samples from Shimla in January 2016 to test for the presence of enterically transmitted hepatitis viruses. Subsequently, 57 icterus patients were tested for various markers of hepatotropic viruses, i.e. anti-HEV IgM/IgG, anti-hepatitis A virus (anti-HAV) IgM/IgG antibodies and HEV RNA. Water samples were screened for HEV and HAV RNA followed by phylogenetic analysis. RESULTS: Overall, 48/57 patients availing municipal water had evidence of HEV infection, detected by serology and RT-PCR. All the water samples tested positive for HEV and HAV RNA, while the patients were negative for anti-HAV IgM antibody, indicating no recent HAV infection. Phylogenetic analysis confirmed the aetiological agent of the current outbreak to be HEV genotype 1. CONCLUSIONS: Serology and RT-PCR confirmed HEV as the aetiology of the outbreak. The absence of new cases of hepatitis A, despite the presence of HAV in the water supply, could be due to previously acquired immunity. Sewage contamination of water leading to faecal-oral transmission of HEV still remains a concern, thus emphasising the need for a vaccination/control strategy.

Immunohistochemistry of immune checkpoint markers PD-1 and PD-L1 in prostate cancer.

Recent availability of immune checkpoint inhibitors has facilitated research involving programmed cell death protein 1 (PD-1) and its ligand, programmed death-ligand 1 (PD-L1). However, the incidence and clinical implication of PD-1 and PD-L1 expression in prostate cancer remain poorly understood. The current study aimed to determine the status of PD-1/PD-L1 expression in prostate cancer specimens and its prognostic significance.We immunohistochemically stained for PD-1 and PD-L1 in our tissue microarray (TMA) consisting of radical prostatectomy specimens. The expression of PD-1/PD-L1 was designated as positive when moderate to strong staining or weak staining was seen in at least 1% or 10%, respectively, of tumor cells and/or associated immune cells. We then evaluated the relationship between the expression of each protein and clinicopathological features available for our patient cohort.PD-1 and PD-L1 were positive in 3 (1.5%) and 1 (0.5%) of 201 non-neoplastic prostate tissues, and also in 17 (7.7%) and 29 (13.2%) of 220 prostate cancers, respectively. PD-1 and PD-L1 were also expressed in tumor-infiltrating lymphocytes/macrophages in 172 (78.2%) and 33 (15.0%) cases, respectively. PD-L1 expression in tumor cells was more often seen in high pT stage (pT2: 10.8% vs pT3/4: 20.4%; P = .072; pT2/3a: 11.4% vs pT3b/4: 31.6%; P = .013) or lymph node-positive (pN0: 10.1% vs pN1: 27.3%; P = .086) cases, whereas PD-1 expression in tumor cells was not significantly associated with pT/pN stage. In addition, there were no statistically significant associations between PD-1/PD-L1 expression in tumor cells or tumor-infiltrating lymphocytes/macrophages versus patient age, preoperative prostate-specific antigen level, or Gleason score. Kaplan-Meier analysis coupled with log-rank test further revealed no significant associations between PD-1/PD-L1 expression in tumor cells (P = .619/P = .315), tumor-infiltrating lymphocytes/macrophages (P = .954/P = .155), or either or both of them (P = .964/P = .767) versus disease recurrence after radical prostatectomy.PD-1/PD-L1 expression was detected in a subset of prostate cancers. In particular, PD-L1 expression was considerably up-regulated in nonorgan-confined tumors. However, PD-1/PD-L1 expression in our TMA was found to be not very helpful in predicting tumor recurrence in prostate cancer patients who underwent radical prostatectomy.

Pneumatosis intestinalis: cost paid for rheumatoid arthritis treatment.

A 75-year-old woman with rheumatoid arthritis on rituximab presented with a 1-week history of constipation and abdominal distension. Subsequent workup showed presence of air in the bowel wall without perforation initially. Due to positive blood cultures, worsening leucocytosis and high suspicion for perforation, an exploratory laparotomy was performed revealing necrotic bowel, walled off perforation and abscess. Patient underwent right hemicolectomy with diversion loop ileostomy. Clinicians must recognise that monoclonal antibodies like rituximab can mask signs of inflammation and therefore should maintain a high index of suspicion for intestinal perforation when evaluating patients with minimal symptoms and pneumatosis intestinalis.

The impact of perivesical lymph node metastasis on clinical outcomes of bladder cancer patients undergoing radical cystectomy.

BACKGROUND: Perivesical lymph nodes (PVLNs) are occasionally isolated during grossing of cystectomy specimens. However, the prognostic implications of the involvement of PVLNs in bladder cancer patients, especially those with comparisons to pN0 disease, remain poorly understood. METHODS: A retrospective review identified 115 radical cystectomy cases where PVLNs had been histologically assessed. These cases were then divided into 4 groups - Group 1 (n = 76): PVLN-negative/other pelvic lymph node (non-PVLN)-negative; Group 2 (n = 5): PVLN-positive/non-PVLN-negative; Group 3 (n = 17): PVLN-negative/non-PVLN-positive; and Group 4 (n = 17): PVLN-positive/non-PVLN-positive. RESULTS: pT stage at cystectomy was significantly higher in Group 3 (P = 0.013), Group 4 (P < 0.001), Groups 2 and 4 (P < 0.001), or Groups 2-4 (P < 0.001) than in Group 1. However, the number of positive PVLNs (mean: 1.8 vs. 2.1; P = 0.718) or the rate of extracapsular extension in the PVLNs (40% vs. 65%, P = 0.609) was not significantly different between Group 2 and Group 4. Kaplan-Meier analysis and log-rank test revealed significantly (P < 0.05) higher risks of disease progression (Group 3/Group 4), cancer-specific mortality (Group 2/Group 3/Group 4), and overall mortality (Group 4), compared with Group 1. Multivariate analysis further showed metastasis to both PVLN and non-PVLN (Group 4), PVLN (Groups 2 and 4), or PVLN and/or non-PVLN (Groups 2-4) as an independent prognosticator for cancer-specific mortality and overall survival. There were also insignificant (P = 0.096) and significant (P = 0.036) differences in cancer-specific survival and overall survival, respectively, between Group 3 versus Group 4, and the trend of the latter was confirmed by subset multivariate analysis (hazard ratio = 3.769; P = 0.099). CONCLUSIONS: Worse prognosis was observed in bladder cancer patients with isolated PVLN metastasis (vs. pN0 disease especially for cancer-specific survival), PVLN metastasis with or without non-PVLN metastasis (vs. pN0 disease), and concurrent PVLN and non-PVLN metastases (vs. PVLN-negative/non-PVLN-positive disease especially for overall survival). These findings indicate the importance of thorough histopathological assessment of PVLNs in radical cystectomy specimens.

Effects of subspecialty signout and group consensus on the diagnosis of microscopic colitis.

Microscopic colitis (MC) includes lymphocytic colitis (LC) and collagenous colitis (CC). Microscopic changes are required to establish these diagnoses. While criteria exist, interobserver variability has been reported previously. This has not been evaluated in the context of subspecialty signout (SSSO) or a consensus conference. We identified 133 colon biopsies diagnosed as LC, CC, MC, or normal but with mild changes insufficient for MC. All predated the introduction of SSSO at our institution. They were independently reviewed by three gastrointestinal (GI) pathologists. Cases lacking independent consensus were reviewed by the same pathologists in consensus conference to establish a final diagnosis. Individual diagnoses were compared with the consensus diagnoses, and consensus diagnoses were compared with original diagnoses made by GI and non-GI pathologists. Consensus diagnoses were normal (n = 34), LC (n = 57), and CC (n = 42). "Normal" was the diagnosis most commonly agreed upon independently (27/34 cases, P = 0.0073 versus LC, P = 0.0172 versus CC). The reviewing pathologists independently agreed with 80%, 80%, and 94% of consensus diagnoses (κ = 0.70, 0.69, and 0.91). The group consensus agreed with the diagnoses in 49 of 58 (84%) cases originally signed out by non-GI pathologists (κ = 0.77) and in 44 of 57 (77%) cases originally signed out by GI pathologists (κ = 0.63). Good interobserver agreement exists for MC, though whether GI subspecialty training improves agreement remains unclear. Group consensus may aid in diagnosis of difficult/borderline MC cases.

Study on Diagnostic Values of Astrocyte Elevated Gene 1 (AEG-1) and Glypican 3 (GPC-3) in Hepatocellular Carcinoma.

OBJECTIVES: To investigate the diagnostic potential of AEG-1 and GPC-3 in hepatocellular carcinoma (HCC). METHODS: AEG-1 and GPC-3 immunohistochemistry were performed on HCC, adjacent nontumor tissue (ANT), and dysplastic nodules (DN). RESULTS: H score of AEG-1 or GPC-3 in HCC was significantly higher than in ANT or DN. In HCC, 92% and 54% showed AEG-1 and GPC-3 positivity, respectively. In ANT, 16.2% were AEG-1 and 7.6% GPC-3 positive. AEG-1 staining was mostly diffuse, whereas GPC-3 frequently showed focal staining. AEG-1 alone showed high sensitivity but low specificity and accuracy. GPC-3, on the other hand, showed high specificity but low sensitivity and accuracy. Combination of both stains boosted the sensitivity, specificity, and accuracy to 94.6%, 89.5%, and 90.5%, respectively, when only diffuse staining was considered as positive. CONCLUSIONS: AEG-1 or GPC-3 alone seemed not an ideal marker for HCC. The combination of AEG-1 and GPC-3 might improve early diagnosis of HCC.

Antibody and Memory B Cell Responses in Hepatitis E Recovered Individuals, 1-30 Years Post Hepatitis E Virus Infection.

Generation and persistence of anti-hepatitis E virus (HEV) antibodies are synonymous with the development of immunity and considered as correlates of protection against HEV infection. However, issues like longevity of immunological memory following recovery from hepatitis E still remains a puzzle. It is critical to understand whether anamnestic response exists for protection from HEV re-infection. The levels and persistence of anti-HEV antibodies were assessed in hepatitis E recovered individuals 1-30 years post HEV infection. The frequencies and functionality of recombinant HEV capsid protein (rORF2p)-stimulated memory B and T cells were also investigated 1-16 years post infection. Anti-HEV antibodies persisted in 91% of hepatitis E recovered individuals. HEV-specific memory B cell responses were detected in 95% of seropositive hepatitis E recovered individuals. CD4+ and CD8+ T cells displayed an effector memory cell phenotype in hepatitis E recovered individuals. In conclusion, long-lived anti-HEV antibodies and HEV-specific memory B cells are maintained for several years in hepatitis E recovered individuals. Involvement of CD4+ and CD8+ effector memory T cells is an important observation since it is inextricably linked to long-lasting protective immunity. In addition to anti-HEV antibodies, possible role of memory B cell response against HEV re-infection could also be considered.

Hepatitis E virus seroprevalence among blood donors in Pune, India.

BACKGROUND: Blood transfusion is a recently reported route of hepatitis E virus (HEV) transmission. It is a bigger concern in regions where large-scale HEV genotype 1 infections occur causing more severe disease. The present study aims to assess the prevalence and rate of HEV infection in the blood donors of Pune, India. MATERIALS AND METHODS: A total of 2447 healthy blood donors were screened for anti-HEV IgG and IgM antibodies. Anti-HEV IgM antibody positives were further subjected to alanine aminotransferase measurement, HEV RNA detection, viral load quantification and phylogenetic analysis. RESULTS: Anti-HEV seroprevalence rate was 17.70%, while IgM prevalence rate was 0.20%. An age dependent increase in IgG seropositive rate was observed. Two of five IgM-positives tested positive for HEV RNA. The viral load ranged from 3.5 × 104 to 4.6 × 105 copies/mL and belonged to HEV genotype 1. CONCLUSIONS: HEV prevalence rate of 17.70% in the blood donors of Pune, India, a developing country, goes at par with the developed countries. Current data of 0.20% (5 of 2447) blood donors positive for anti-HEV IgM and two of them being HEV RNA positive suggest a need for consideration of cost-effective evaluation towards pooled HEV RNA testing in blood banks.

Percent Gleason pattern 4 in stratifying the prognosis of patients with intermediate-risk prostate cancer.

The Gleason score remains the most reliable prognosticator in men with prostate cancer. One of the recent important modifications in the Gleason grading system recommended from the International Society of Urological Pathology consensus conference is recording the percentage of Gleason pattern 4 in the pathology reports of prostate needle biopsy and radical prostatectomy cases with Gleason score 7 prostatic adenocarcinoma. Limited data have indeed suggested that the percent Gleason pattern 4 contributes to stratifying the prognosis of patients who undergo radical prostatectomy. An additional obvious benefit of reporting percent pattern 4 includes providing critical information for treatment decisions. This review summarizes and discusses available studies assessing the utility of the percentage of Gleason pattern 4 in the management of prostate cancer patients.

Life-Saving Combined Heart-Kidney Transplantation in a Previous Sequential Heart and Kidney Transplant Recipient.

PURPOSE: Solid organ re-transplantation in the context of allograft failure is a challenging clinical and ethical problem. Ideally, solid organ re-transplantation after initial allograft failure should be performed in all recipients, but this is often not clinically or logistically feasible. METHODS: This report details what we believe is the first combined heart-kidney transplant in a recipient of a previous sequential heart and kidney transplant. RESULTS: Eight years after a combined heart and kidney transplant after initially receiving a sequential heart and kidney transplant, a 31-year-old man is doing extremely well, with no rejection episodes or significant complications after transplantation. SUMMARY: This case confirms that combined heart and kidney transplantation is a viable option for tackling the complex issue of graft failure in recipients of previous cardiac and renal grafts. LEARNING POINTS: Good short- and long-term outcomes following combined heart-kidney transplantation can be achieved in patients with multi-system end-organ dysfunction.Advances in immunosuppressant therapy have enabled multiple transplantation procedures from different donors in a single recipient.We describe the first recipient in the world of combined heart-kidney transplantation on a background of previous sequential heart-kidney transplantation.

Impaired NK cell functionality and increased TNF-α production as biomarkers of chronic chikungunya arthritis and rheumatoid arthritis.

The chronic chikungunya arthritis symptoms closely mimic the rheumatoid arthritis (RA) symptoms, thus making it difficult to distinguish between these two clinical entities. The current comparative study characterizes NK (CD3-CD56+) and NK-like T (CD3+CD56+) cell responses in patients with chronic chikungunya arthritis and RA. Phenotype and functions of NK and NK-like T cells repertoire were assessed in 56 chronic chikungunya arthritis, 26 RA patients and 82 controls using flow cytometry. TNF-α and IFN-γ-secreting NK-like T cells were high in both chronic arthritis patients than in controls. Percentage of TNF-α+ NK cells was higher in RA patients than in controls. Percentage of perforin+ NK cells was low in both chronic arthritis patient groups. Among the patient groups, expressions of perforin+ and IFN-γ+ NK-like T cells were higher in RA. Overall, our data show reduced frequency of NK-like T cells, lower expression of perforin+ NK, higher expression of TNF-α+ NK-like T and IFN-γ+ NK-like T cells as the markers of chronic arthritic diseases. In the absence of any specific treatment for chronic chikungunya induced arthritis and promising results of anti-TNF-α therapy against RA, current data may form the basis for future in vivo studies and has scope as possible therapeutics against chikungunya.