New-onset Diabetes Mellitus after EUS-guided Drainage with LAMS: A Pilot Study.

BACKGROUND AND AIMS: Walled-off necrosis (WON) is a serious complication of severe pancreatitis, patients with necrotizing pancreatitis having an increased risk of developing diabetes mellitus (DM). The aim of this study was to assess the frequency of new-onset diabetes (NOD) in patients with symptomatic WON after endoscopic ultrasound (EUS)-guided drainage with lumen-apposing metal stents (LAMS). METHODS: We retrospectively analyzed a prospectively collected database of patients with symptomatic WON treated by EUS-guided drainage with LAMS in a tertiary referral center. The patients were followed-up for at least 12 months after stent removal. These patients were compared with age- and sex-matched asymptomatic WON controls without interventional treatment and healthy controls to assess the one-year occurrence of DM. Diabetes was defined according to the American Diabetes Association criteria. RESULTS: Of the 50 patients with symptomatic WON included in the study (male/female ratio, 33:17; median age, 60 years), 13 patients (26%) had pre-existing DM and were excluded. Ten of the remaining 37 patients (27%) without prior DM developed NOD within one year after stent removal, this frequency being higher than in asymptomatic WON controls (18.9%, p=0.581) and healthy controls (2%, p = 0.002). In the symptomatic WON group, NOD patients compared to non-DM patients were older (63.5 vs. 56 years old, p=0.042), had more frequent necrosis > 50% of the pancreatic parenchyma (p=0.002) and had a body-tail location of WON (p<0.001). On multivariate analysis, the number of direct endoscopic necrosectomy (DEN) sessions was the only significant factor for NOD occurrence (OR=7.05, p=0.010). NOD patients had poor glycemic control and required more DEN sessions to achieve WON resolution than patients with prior DM (p=0.017). CONCLUSIONS: In patients with symptomatic WON treated by EUS-guided drainage, DM occurred in 27% of previously non-diabetic patients within one year of follow-up. Patients with extensive pancreatic necrosis were more likely to develop NOD, a high number of DEN sessions being a significant risk factor for NOD occurrence.

Activated Hepatic Stellate Cells (Ito Cells) - Marker of Advanced Fibrosis in Chronic Viral Hepatitis C: A Pilot Study.

BACKGROUND AND AIMS: The aim of this study is to determine whether activated hepatic stellate cells (HSCs) may represent a prognostic marker of progressive liver fibrosis in chronic viral hepatitis C (VHC) before antiviral therapy. The possible correlation between HSCs immunohistochemical features, histopathological aspects and clinical data before therapy were also studied. METHODS: This retrospective pilot study was conducted on 27 liver biopsies from VHC patients before antiviral therapy. HSCs's immunohistochemical analysis used the antibodies alpha-smooth muscle actin (α-SMA), glial fibrillary acidic protein (GFAP) and vinculin. We correlated immunopositive HSCs with HCV load, liver stiffness (LS), fibrosis stage and necro-inflammatory degree before treatment. Also, we assessed the association between liver fibrosis after therapy, the sustained virological response at 12 weeks after therapy (SVR 12) and the type of therapy. RESULTS: HSCs were increased in VHC patients compared to controls, mainly in the intermediate and periportal lobular regions. α-SMA and vinculin HSCs correlated positively with fibrosis stage (p=0.044), (p=0.028). Furthermore, α-SMA and vinculin HSCs were associated with LS (p=0.027), (p=0.002) and viral load (p=0.021), (p=0.006), but not with necro-inflammation degree. GFAP HSCs inversely correlated with fibrosis stage (r= -0.475), LS (r= -0.422) and HCV load (r= -0.517), but positively with necro-inflammation degree (p=0.038). Liver fibrosis post therapy correlated positively with SVR12 (p<0.001) and the type of therapy (p=0.006) and SVR12 correlated positively with treatment's type (p=0.002). CONCLUSIONS: Activated HSCs may represent a marker of increased liver fibrosis in VHC. Different immunohistochemical markers can detect various HSCs subpopulations involved in the evolution of VHC and liver fibrosis.

External Validation of Mortality Scores among High-Risk COVID-19 Patients: A Romanian Retrospective Study in the First Pandemic Year.

BACKGROUND: We aimed to externally validate three prognostic scores for COVID-19: the 4C Mortality Score (4CM Score), the COVID-GRAM Critical Illness Risk Score (COVID-GRAM), and COVIDAnalytics. METHODS: We evaluated the scores in a retrospective study on adult patients hospitalized with severe/critical COVID-19 (1 March 2020-1 March 2021), in the Teaching Hospital of Infectious Diseases, Cluj-Napoca, Romania. We assessed all the deceased patients matched with two survivors by age, gender, and at least two comorbidities. The areas under the receiver-operating characteristic curves (AUROCs) were computed for in-hospital mortality. RESULTS: Among 780 severe/critical COVID-19 patients, 178 (22.8%) died. We included 474 patients according to the case definition (158 deceased/316 survivors). The median age was 75 years; diabetes mellitus, malignancies, chronic pulmonary diseases, and chronic kidney and moderate/severe liver diseases were associated with higher risks of death. According to the predefined 4CM Score, the mortality rates were 0% (low), 13% (intermediate), 27% (high), and 61% (very high). The AUROC for the 4CM Score was 0.72 (95% CI: 0.67-0.77) for in-hospital mortality, close to COVID-GRAM, with slightly greater discriminatory ability for COVIDAnalytics: 0.76 (95% CI: 0.71-0.80). CONCLUSION: All the prognostic scores showed close values compared to their validation cohorts, were fairly accurate in predicting mortality, and can be used to prioritize care and resources.

Treatment with Tocilizumab in Adult Patients with Moderate to Critical COVID-19 Pneumonia: A Single-Center Retrospective Study.

OBJECTIVES: This study aimed to assess if tocilizumab (TCZ) timing is associated with improved survival. MATERIAL AND METHODS: Data obtained from adult patients with moderate/severe/critical COVID-19 and treated with TCZ, who were admitted to the Teaching Hospital of Infectious Diseases, Cluj-Napoca, Romania (April 2020-April 2021), were retrospectively analyzed. The database included demographics, clinical data, computed tomography scan results, the kinetics of IL-6, laboratory variables, and the outcome until discharge. RESULTS: A total of 221 patients received dexamethasone, antivirals, anticoagulants, and 1-2 doses of TCZ, 8 mg/kg. In 2021, more patients received high-flow oxigen/non-invasive ventilation compared to those hospitalized in 2020, but demographics, in-hospital mortality, and laboratory data did not differ significantly. In-hospital mortality was associated with age, disease severity, lung damage, intensive care unit (ICU) admission, cardiovascular comorbidities, and IL-6>100 pg/mL at TCZ administration. In multivariate analysis the risk of death was significantly higher in patients with a persistent inflammatory state, adjusted odds ratio (aOR) 16.6 (95% CI 3.07-108.96); lung damage>40%, aOR 11.68 (95% CI 2.05-224.98); and cardiovascular comorbidities>2, aOR 3.65 (95% CI 1.06-12.53). TCZ initiation at ≤3 days after admission showed improved survival, odds ratio (OR)=0.39 (95% CI 0.16-0.9). Severe infections were found in 11 (4.9%) patients. CONCLUSION: Early initiation of TCZ seems beneficial and safe in patients with moderate to critical COVID-19 pneumonia.

Antibody and viral RNA kinetics in SARS-CoV2 infected patients admitted to a Romanian University Hospital of Infectious Diseases.

OBJECTIVES: To assess the antibody and viral kinetics in asymptomatic/mild confirmed SARS-CoV-2 infections compared to more severe patients. MATERIAL AND METHODS: Retrospective analysis of data obtained from adult patients with a confirmed SARS-CoV2 infection having at least one SARS-CoV-2 pair of specific IgM/IgG tests, admitted in The University Hospital of Infectious Diseases Cluj-Napoca, Romania (28 February to 31 August 2020). The database also included: demographic, clinical, chest X-ray and/or CT scan results, RT-PCR SARS-CoV-2, and dexamethasone treatment. A total of 469 patients were evaluated as "asymptomatic/mild" and "moderate/severe/critical" cases. RESULTS: The median time since confirmation to SARS-CoV-2 PCR negativity was 15 days [95% CI: 13-18] in asymptomatic/mild cases and 17 days [95% CI: 16-21] in moderate/severe ones. The median time to seroconversion for both IgM and IgG was 13 days [95% CI: 13-14] in asymptomatic/mild cases and 11 days [95% CI: 10-13] in moderate/severe ones. For both antibody types, the highest reactivity was significantly associated with more severe presentation (IgM: OR = 10.30, IgG: OR = 7.97). CONCLUSION: Asymptomatic/mild COVID-19 cases had a faster RT-PCR negativity rate compared to moderate/severe/critical patients. IgG and IgM dynamics were almost simultaneous, more robust for IgG in more severe cases, and at one month after confirmation, almost all patients had detectable antibody titers.

Role of Adiponectin and Tumor Necrosis Factor-Alpha in the Pathogenesis and Evolution of Type 1 Diabetes Mellitus in Children and Adolescents.

Type 1 diabetes mellitus (T1DM) is a complex condition caused by the destruction of pancreatic beta cells by autoimmune mechanisms. As a result, insulin deficiency and subsequent hyperglycemia occur. The aim of the present study is to investigate the role of adiponectin and tumor necrosis factor alpha (TNF-α) in the development of T1DM. The study is designed as an observational case-control study, involving 52 diabetic patients and 66 controls. Z scores for Body Mass Index (BMI), weight, height, and adiponectin and TNF-α serum levels were assessed in both groups. The T1DM group had significantly higher TNF-α levels and a significantly higher proportion of high-risk patients for inflammation based on TNF-α values as compared to the control group, while both groups had statistically similar adiponectin levels and a similar proportion of high/medium-risk patients based on adiponectin values. TNF-α plays a significant role in the pathogenesis and evolution of T1DM and it may represent an additional marker of disease progression, as well as a potential target of immunotherapeutic strategies. In the present study, no statistically significant differences were recorded in adiponectin levels neither in diabetic patients and controls, nor in high/medium severity risk diabetic patients.

Variable anti-HBs antibody titers in vaccinated birth cohorts - A cross-sectional population-based study.

BACKGROUND: After the introduction of hepatitis B (HB) vaccination in 1995 in newborns, two catch-up campaigns targeted unvaccinated 9 year old in 2000-2003 (born 1991-1994) and the 18 year old in 2004-2008 (born 1986-1990), resulting in several birth-cohorts. Our objective was to assess the anti-HBs titers in each birth-cohort. METHODS: We included all outpatients (78.5%) and hospitalized patients with measured anti-HBs antibody titers in the Teaching Hospital of Infectious Diseases, Cluj-Napoca, Romania, during April 2014 - December 2018 (without HB history). We compared the anti-HBs titers in all birth-cohorts using the Lexis surfaces (titers by age, time period and cohort patterns). We also evaluated the number of acute HB in the corresponding inpatient birth-cohorts and special groups. RESULTS: We included 2963 participants, mean age = 31.0 ± 14.2, 64.1% women. The birth-cohort 1995-2006, vaccinated after delivery (n = 424, 3-dose HB vaccine coverage > 90%), had significantly lower protective titers (41.3% >10 mIU/mL) compared to the other birth-cohorts: born after 2007 (also vaccinated at birth, 67.0%, n = 106), 1991-1994 (age 9, 74.3%, n = 847), 1986-1990 (age 18, 71.3%, n = 543). In the unvaccinated cohort (n = 1043, mean age = 45.5 ± 12.4) protective titers were found in 44.8%, probably after self-limited HB infection. Concordant results were found using the proportion of patients with detectable or robust titers, and median or geometric mean titers. Four breakthrough acute HB infections were hospitalized of the corresponding vaccinated cohorts (birth years 1988, 1990, 1995, 1996). Data on a few tested infants (n = 47, not included in the main study) demonstrated good protection, 88.9%. CONCLUSIONS: Our study demonstrated the long-term evidence of protection of HBV vaccine at two decades following the primary immunization and a booster seems unsupported. Further studies should be done to assess the need of a booster dose within the general population and special groups.

A comparison of hepatitis E and A in a teaching hospital in Northwestern Romania. Acute hepatitis E - a mild disease?

BACKGROUND AND AIMS: The incidence of locally acquired hepatitis E has increased in recent years across Europe. There are only few data on hepatitis E in Romania. The purpose of our research was to describe and compare hepatitis E and hepatitis A in adult patients. METHODS: We included all consecutive adult patients with hepatitis E and hepatitis A admitted to the Teaching Hospital of Infectious Diseases, Cluj-Napoca, Romania between January 2017 and August 2019. RESULTS: Hepatitis E incidence increased in 2018-2019 compared to 2017. The average age in hepatitis E (n=48) patients was 50.6 versus 39.1 years in hepatitis A (n=152, not including 262 minors) and two-thirds of the patients in both groups were men. Compared to hepatitis A, patients with hepatitis E presented significantly less modified AST and ALT, bilirubin, prothrombin index and INR levels. We found more comorbidities in hepatitis E patients adjusted for age and gender. Severe forms were found in 5 (3.3%) hepatitis A patients, compared to 12 (25%) of hepatitis E patients, of which 3 died. Ribavirin treatment was considered in 9 patients with acute-on-chronic hepatitis E, immunosuppression, cancers or neurological manifestations, showing good results. CONCLUSIONS: We observed an increased number of hepatitis E cases. Although laboratory results were less modified compared to hepatitis A, we found a higher number of severe hepatitis E cases. Ribavirin treatment seems to be beneficial in patients with preexisting conditions.

Changes observed in erythrocyte cells exposed to an alternating current.

BACKGROUND AND AIMS: Appliance of electric pulses induces red blood cells (RBCs) membrane poration, membrane aminophospholipid perturbation and alteration of the normal flip-flop process, resulting in various shape changes of the RBCs. We studied morphological and water permeability changes of RBCs bombarded with electrons in an alternating current circuit. METHODS: We used three venous blood samples of 100 mL and an alternating current device. The harvested blood was divided into four experimental sets to be used for various exposure times: 0 hours (control RBCs), 0.5h, 3h and 6h (electric-stimulated RBCs). Following the electric current each of the four sets were further divided into three samples: one for the assessment of the echinocytes/RBCs ratio, another for the electron microscopy study of ultrastructural changes induced by the alternating electrical current and a larger third one for determining water permeability of RCBs by 1H-NMR spectroscopy and morphological measurements. RESULTS: There is a small but statistically significant effect of the RBC exposure to alternating electric current on cell diameters. Exposure to electric current is positively and strongly correlated with the percentage of echinocytes in a duration-dependent manner. There is a strong and statistically significant correlation between electric current exposure and permeability to water as measured by 1H-NMR spectroscopy. CONCLUSION: Following interactions between electric current and RBC membrane, certain modifications were observed in the erythrocyte structure. We attribute the increased cell size to a higher permeability to water and a decreased tonicity. This leads to the transformation of the RBCs into echinocytes.

The ubiquitin proteasomal system: a potential target for the management of Alzheimer's disease.

The cellular quality control system degrades abnormal or misfolded proteins and consists of three different mechanisms: the ubiquitin proteasomal system (UPS), autophagy and molecular chaperones. Any disturbance in this system causes proteins to accumulate, resulting in neurodegenerative diseases such as amyotrophic lateral sclerosis, Alzheimer's disease (AD), Parkinson's disease, Huntington's disease and prion or polyglutamine diseases. Alzheimer's disease is currently one of the most common age-related neurodegenerative diseases. However, its exact cause and pathogenesis are unknown. Currently approved medications for AD provide symptomatic relief; however, they fail to influence disease progression. Moreover, the components of the cellular quality control system represent an important focus for the development of targeted and potent therapies for managing AD. This review aims to evaluate whether existing evidence supports the hypothesis that UPS impairment causes the early pathogenesis of neurodegenerative disorders. The first part presents basic information about the UPS and its molecular components. The next part explains how the UPS is involved in neurodegenerative disorders. Finally, we emphasize how the UPS influences the management of AD. This review may help in the design of future UPS-related therapies for AD.

Role of toll-like receptors in multiple myeloma and recent advances.

Multiple myeloma (MM) is a hematologic malignancy characterized as an abnormal proliferation and invasion of plasma cells into the bone marrow. Toll-like receptors (ТLRs) connect the innate and adaptive immune responses and represent a significant and potentially linking element between inflammation and cancer. When TLRs bind to their ligands, they trigger two major signaling pathways such that both share overlapping downstream signals: one is a myeloid differentiation primary response 88 (MyD88)-dependent production and activation of nuclear factor-κB, whereas the other is a MyD88-independent production of type-I interferon. Whereas the MyD88 pathway results in proinflammatory cytokine production, the other pathway stimulates cell proliferation. Dysregulations of these pathways may eventually lead to abnormal cell proliferation and MM. Despite recent biomedical advances, MM continues to be an incurable disease. There are an increasing number of TLR-based therapeutic approaches currently being tested in a number of preclinical and clinical studies. We here attempt to outline in detail the currently available information on TLRs in various types of cancer.