Impact of bound ssRNA length on allostery in the Dengue Virus NS3 helicase.

The presence of ATP is known to stimulate helicase activity of the Dengue Virus Non-structural protein 3 helicase (NS3h), and the presence of RNA stimulates NS3h ATPase activity, however this coupling is still mechanistically unclear. Here we use atomistic models and molecular dynamics simulations to evaluate the single-stranded RNA (ssRNA)-length dependence of the NS3h-ssRNA binding affinity and its modulation by bound ATP. Considering complexes with 7, 11, 16 and 26 nucleotides (nts), we observe that both the binding affinity and its modulation by bound ATP are augmented with increased ssRNA lengths.

Thermodynamic and mechanistic analysis of the functional properties of dengue virus NS3 helicase.

The Dengue Virus (DENV) non-structural protein 3 (NS3) is a multi-functional protein critical in the viral life cycle. The DENV NS3 is comprised of a serine protease domain and a helicase domain. The helicase domain itself acts as a molecular motor, either translocating in a unidirectional manner along single-stranded RNA or unwinding double-stranded RNA, processes fueled by the hydrolysis of nucleoside triphosphates.

Effects of heat stress on sperm quality of French Bulldogs.

This study aimed to evaluate the effects of heat stress (HS) and its duration on semen quality, serum testosterone, pulsatility and resistibility index of the testicular artery of French Bulldogs. Eight male French Bulldogs, 3-7 years old, 12.63 ± 1.

Insights into the product release mechanism of dengue virus NS3 helicase.

The non-structural protein 3 helicase (NS3h) is a multifunctional protein that is critical in RNA replication and other stages in the flavivirus life cycle. NS3h uses energy from ATP hydrolysis to translocate along single stranded nucleic acid and to unwind double stranded RNA. Here we present a detailed mechanistic analysis of the product release stage in the catalytic cycle of the dengue virus (DENV) NS3h.

Atomistic insight into the essential binding event of ACE2-derived peptides to the SARS-CoV-2 spike protein.

The pathogenic agent of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) enters into human cells through the interaction between the receptor binding domain (RBD) of its spike glycoprotein and the angiotensin-converting enzyme 2 (ACE2) receptor. Efforts have been made towards finding antivirals that block this interaction, therefore preventing infection. Here, we determined the binding affinity of ACE2-derived peptides to the RBD of SARS-CoV-2 experimentally and performed MD simulations in order to understand key characteristics of their interaction.

Monoclonal free light chain detection and quantification: Performances and limits of available laboratory assays.

The detection and quantification of immunoglobulin free light chains in serum and urine is recommended for the diagnosis and monitoring of monoclonal gammopathies according to the guidelines of the International Myeloma Working Group (IMWG). Several tests are currently available in the clinical laboratory to detect and quantify free light chains but although quality, efficiency, and effectiveness have been improved, the results are still variable and poorly harmonized and standardized. The present review article wants to analyze these aspects, with a keen eye on techniques, such as mass spectrometry, that could replace in the practical clinical laboratory the current methods including Bence-Jones protein assay and free light chain immunoassays.

A significant decreased in the frequency of Clostridioides difficile infection at the Italian Hospital of Desio over the last decade.

Clostridiodes difficile infection (CDI) is the most important cause of healthcare-associated diarrhea. The decreasing trend of CDI from 15% to 4% observed at the Italian Hospital of Desio over a 10-year period is due to prevention strategies. Our data highlight the importance of surveillance studies to control CDI.

Baseline characteristics of COVID-19 Italian patients admitted to Desio Hospital, Lombardy: a retrospective study.

The correlation of clinical, radiological and laboratory findings of patients at admission in the Emergency Department (ED) with clinical severity and risk of mortality was investigated. Adult coronavirus disease 2019 (COVID-19) patients hospitalized in March 2020 in Desio Hospital, Lombardy, were retrospectively included in the study, and categorized in terms of disease severity and adverse outcome. Out of the 175 patients enrolled, 79% presented one or more comorbidities, with cardiovascular disease being the most frequent (62%).

A rare case of Clostridium paraputrificum bacteremia in a 78-year-old Caucasian man diagnosed with an intestinal neoplasm.

Clostridium like species, particularly Clostridium perfrigens, are the second most common causes of human anaerobic infections, including myonecrosis and bacteremia. Clostridium paraputrificum is an infrequent isolate, which has been identified in only 1% of reported cases of clostridial infections. We herein report a rare case of C.

A rare case of cutaneous Papiliotrema (Cryptococcus) laurentii infection in a 23-year-old Caucasian woman affected by an autoimmune thyroid disorder with hypothyroidism.

In recent years, the frequency of infections due to saprophytic fungi has increased. Cryptococcus laurentii, recently classified as Papiliotrema laurentii, is responsible for fungemia, meningitis, and superficial infections. Here, we report the first case of cutaneous Papiliotrema (Cryptococcus) laurentii infection in a 23-year-old Caucasian woman affected by an autoimmune thyroiditis with hypothyroidism.

Nucleotide-dependent dynamics of the Dengue NS3 helicase.

Dengue represents a substantial public health burden, particularly in low-resource countries. Non-structural protein 3 (NS3) is a multifunctional protein critical in the virus life cycle and has been identified as a promising anti-viral drug target. Despite recent crystallographic studies of the NS3 helicase domain, only subtle structural nucleotide-dependent differences have been identified, such that its coupled ATPase and helicase activities remain mechanistically unclear.

Rapid Identification of Carbapenemase-producing strains by Matrix-Assisted Laser Desorption/Ionization-Time of Flight using Vitek Mass Spectrometry System.

Objective: The analysis of the protein pattern of carbapenemase (KPC)-producing strains by Bruker Matrix-Assisted Laser Desorption Ionization (MALDI) Biotyper system has revealed the presence, in the majority of cases, of an 11.109 peak. The peak corresponds to the gene product named p019 of the -bearing plasmids and has been suggested as a candidate for a biomarker that is able to distinguish KPC-producers from non-KPC-producers.

Multiple Parasitic Infestation in a Nine-month-old Patient: A Case Report.

We are reporting the case of a nine-month-old Pakistani female with complaint of growth retardation who presented multiple intestinal parasitic infections. Probably because of contamination with fecal matter, the initial microscopic examination of the urinary sample revealed the presence of eggs of , cysts of , and an organism similar to mites. Stool samples were obtained after two weeks and microscopic investigation confirmed the presence of eggs, cysts of , and hookworm eggs.

In Vitro Activity of Antifungal Drugs Against Trichophyton rubrum and Trichophyton mentagrophytes spp. by E-Test Method and Non-supplemented Mueller-Hinton Agar Plates.

Trichophyton rubrum and Trichophyton mentagrophytes spp. are two of the most frequently isolated dermatophytes causing dermatophytosis worldwide. Since the incidence of resistance to antifungal agents is increasing, antifungal susceptibility tests are needed to successfully treat dermatophytoses.

[Monoclonal gammopathy of uncertain significance (MGUS) and occupational risk factors: criteria to carry out the health surveillance].

Monoclonal gammopathy of uncertain significance (MGUS) identifies a clinically asymptomatic and laboratory-based situation characterized by a modest monoclonal component (MC). In a limited percentage of cases, on a probabilistic basis, the asymptomatic genepremalignant stage could lead to multiple myeloma (MM). Materials and Methods.

The importance of considering the neglected intestinal protozoan parasite Dientamoeba fragilis.

Dientamoebiasis is globally distributed and detected in a large number of subjects with diarrhea, abdominal discomfort, flatulence, fatigue and loss of appetite. The life cycle and transmission of Dientamoeba fragilis are poorly understood. Microscopic examination of permanent stained smears is traditionally employed to diagnose the infection.

Slow magnetization dynamics in Co(ii)/Co(iii) triethanolamine/pivalate complexes.

We report the synthesis, structural characterization and a combined computational and experimental study of the magnetic properties of two pivalate cobalt complexes, a mononuclear Co(ii) one and a tetranuclear Co(ii)Co(iii) mixed valence polynuclear one. The latter shows SMM behaviour revealed under an applied DC field with a thermal barrier of ca. 30 cm competing with direct and Raman relaxation processes.

Genus-level identification of dermatophytes by MALDI-TOF MS after 2 days of colony growth.

Unlabelled: Matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS) is becoming a popular technology in clinical microbiology. It is a fast and highly specific method for the routine identification of micro-organisms. In this study, we evaluated the suitability of dermatophyte identification after only 2 days of colony growth using MALDI-TOF MS.

IgMκ-IgMλ pair quantitation in the clinical laboratory practice.

Background: New Hevylite® assay quantifies the immunoglobulin classes, including IgM bound to light chains, allowing distinguishing immunoglobulins involved and uninvolved in plasma cell disorders.

Objective: To compare data obtained by IgM Hevylite® (IgM-HLC) assay with conventional methods used in routine laboratory practice for monitoring IgM plasma cell disorders.

Methods: Serum samples (n=122) from 50 patients with IgM monoclonal protein (MP) identified by Immunofixation (IFE) before the beginning of the study were collected during monitoring from December 2012 to September 2014 (2 Waldestrom's macroglobulinemia, 4 NH-lymphoma, 44 MGUS) and were assessed using IgM Hevylite® (HLC) assay, Capillary Electrophoresis (CE), Immunofixation (IFE), serum Free Light Chain (FLC) assay and total IgM measurements.

Proteomic analysis in clear cell renal cell carcinoma: identification of differentially expressed protein by 2-D DIGE.

Renal cell carcinoma (RCC), the most common neoplasm affecting the adult kidney, is characterised by heterogeneity of histological subtypes, drug resistance, and absence of molecular markers. Two-dimensional difference gel electrophoresis (2-D DIGE) technology in combination with mass spectrometry (MS) was applied to detect differentially expressed proteins in 20 pairs of RCC tissues and matched adjacent normal kidney cortex (ANK), in order to search for RCC markers. After gel analysis by DeCyder 6.

Perinatal exposure to low doses of dioxin can permanently impair human semen quality.

Background: In recent decades, young men in some industrialized areas have reportedly experienced a decrease in semen quality.

Objective: We examined effects of perinatal dioxin exposure on sperm quality and reproductive hormones.

Methods: We investigated sperm quality and hormone concentrations in 39 sons (mean age, 22.

Dioxin exposure, from infancy through puberty, produces endocrine disruption and affects human semen quality.

Background: Environmental toxicants are allegedly involved in decreasing semen quality in recent decades; however, definitive proof is not yet available. In 1976 an accident exposed residents in Seveso, Italy, to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD).

Objective: The purpose of this study was to investigate reproductive hormones and sperm quality in exposed males.

Differential expression of AQP1 in microdomain-enriched membranes of renal cell carcinoma.

Human aquaporin-1 (AQP1) is the most studied member of the aquaporin family, acting as molecular water channel. It is also considered a differentiation marker for proximal renal tubular cells, from which clear cells renal cell carcinoma (RCC) originates, playing an important role in urine formation. We therefore studied AQP1 expression at the proteomic level in RCC and normal tissues, mainly focusing on microdomain-enriched membranes in which AQP1 is highly concentrated.