Kinetic Study of Laboratory Mutants of NDM-1 Metallo-beta-Lactamase and the Importance of an Isoleucine at Position 35.

peer reviewedTwo laboratory mutants of NDM-1 were generated by replacing the isoleucine at position 35 with threonine and serine residues: the NDM-1(I35T)and NDM-1(I35S)enzymes. These mutants were well characterized, and their kinetic parameters were compared with those of the NDM-1 wild type. Thekcat,Km, andkcat/Kmvalues calculated for the two mutants were slightly different from those of the wild-type enzyme. Interestingly, thekcat/Kmof NDM-1(I35S)for loracarbef was about 14-fold higher than that of NDM-1. Far-UV circular dichroism (CD) spectra of NDM-1 and NDM-1(I35T)and NDM-1(I35S)enzymes suggest local structural rearrangements in the secondary structure with a marked reduction of alpha-helix content in the mutants

Dose-Dependent Impairment of the Immune Response to the Moderna-1273 mRNA Vaccine by Mycophenolate Mofetil in Patients with Rheumatic and Autoimmune Liver Diseases

The purpose of this study was to evaluate the efficacy and safety of the Moderna-1273 mRNA vaccine for SARS-CoV-2 in patients with immune-mediated diseases under different treatments. Anti-trimeric spike protein antibodies were tested in 287 patients with rheumatic or autoimmune diseases (10% receiving mycophenolate mofetil, 15% low-dose glucocorticoids, 21% methotrexate, and 58% biologic/targeted synthetic drugs) at baseline and in 219 (76%) 4 weeks after the second Moderna-1273 mRNA vaccine dose. Family members or caretakers were enrolled as the controls. The neutralizing serum activity against SARS-CoV-2-G614, alpha, and beta variants in vitro and the cytotoxic T cell response to SARS-CoV-2 peptides were determined in a subgroup of patients and controls. Anti-SARS-CoV-2 antibody development, i.e., seroconversion, was observed in 69% of the mycophenolate-treated patients compared to 100% of both the patients taking other treatments and the controls (p < 0.0001). A dose-dependent impairment of the humoral response was observed in the mycophenolate-treated patients. A daily dose of >1 g at vaccination was a significant risk factor for non-seroconversion (ROC AUC 0.89, 95% CI 0.80-98, p < 0.0001). Moreover, in the seroconverted patients, a daily dose of >1 g of mycophenolate was associated with significantly lower anti-SARS-CoV-2 antibody titers, showing slightly reduced neutralizing serum activity but a comparable cytotoxic response compared to other immunosuppressants. In non-seroconverted patients treated with mycophenolate at a daily dose of >1 g, the cytotoxic activity elicited by viral peptides was also impaired. Mycophenolate treatment affects the Moderna-1273 mRNA vaccine immunogenicity in a dose-dependent manner, independent of rheumatological disease

Rubinstein-Taybi Syndrome: spectrum of CREBBP mutations in Italian patients

BACKGROUND: Rubinstein-Taybi Syndrome (RSTS, MIM 180849) is a rare congenital disorder characterized by mental and growth retardation, broad and duplicated distal phalanges of thumbs and halluces, facial dysmorphisms and increased risk of tumors. RSTS is caused by chromosomal rearrangements and point mutations in one copy of the CREB-binding protein gene (CREBBP or CBP) in 16p13.3. To date mutations in CREBBP have been reported in 56.6% of RSTS patients and an average figure of 10% has ascribed to deletions. METHODS: Our study is based on the mutation analysis of CREBBP in 31 Italian RSTS patients using segregation analysis of intragenic microsatellites, BAC FISH and direct sequencing of PCR and RT-PCR fragments. RESULTS: We identified a total of five deletions, two of the entire gene and three, all in a mosaic condition, involving either the 5' or the 3' region. By direct sequencing a total of 14 de novo mutations were identified: 10 truncating (5 frameshift and 5 nonsense), one splice site, and three novel missense mutations. Two of the latter affect the HAT domain, while one maps within the conserved nuclear receptor binding of (aa 1–170) and will probably destroy a Nuclear Localization Signal. Identification of the p.Asn1978Ser in the healthy mother of a patient also carrying a de novo frameshift mutation, questions the pathogenetic significance of the missense change reported as recurrent mutation. Thirteen additional polymorphisms, three as of yet unreported, were also detected. CONCLUSION: A high detection rate (61.3%) of mutations is confirmed by this Italian study which also attests one of the highest microdeletion rate (16%) documented so far

MORFEO enters final design phase

MORFEO (Multi-conjugate adaptive Optics Relay For ELT Observations, formerly MAORY), the MCAO system for the ELT, will provide diffraction-limited optical quality to the large field camera MICADO. MORFEO has officially passed the Preliminary Design Review and it is entering the final design phase. We present the current status of the project, with a focus on the adaptive optics system aspects and expected milestones during the next project phase

PPAR agonists for the treatment of primary biliary cholangitis: Old and new tales

Introduction: Primary biliary cholangitis (PBC) is an autoimmune liver disease involving the small intrahepatic bile ducts; when untreated or undertreated, it may evolve to liver fibrosis and cirrhosis. Ursodeoxycholic Acid (UDCA) is the standard of care treatment, Obeticholic Acid (OCA) has been approved as second-line therapy for those non responder or intolerant to UDCA. However, due to moderate rate of UDCA-non responders and to warnings recently issued against OCA use in patients with cirrhosis, further therapies are needed.Areas covered. Deep investigations into the pathogenesis of PBC is leading to proposal of new therapeutic agents, among which peroxisome proliferator-activated receptor (PPAR) ligands seem to be highly promising given the preliminary, positive results in Phase 2 and 3 trials. Bezafibrate, the most evaluated, is currently used in clinical practice in combination with UDCA in referral centers. We herein describe completed and ongoing trials involving PPAR agonists use in PBC, analyzing pits and falls. Expert opinion: Testing new therapeutic opportunities in PBC is challenging due to its low prevalence and slow progression. However, new drugs including PPAR agonists, are currently under investigation and should be considered for at-risk PBC patients

Evaluation of Commercial Meat Products of Red Chicken Reared under LED Lights

The objective of our study was to investigate the role of three different light-color temperatures of Light-Emitting Diodes (LEDs) [Neutral (K=3300−3700); Warm (K=3000−2500) and Cool (K=5500−6000)] on the qualitative attributes of breast meat obtained from male AZ Extra Heavy Red chickens. The comparison was made with meat deriving from chickens reared in the presence of classic neon lighting (Control). The meat was analyzed for the determination of both physical and chemical properties (cooking loss, moisture, total lipids and fatty acid composition). Furthermore, meat samples subjected to cooking were also analyzed for the identification of volatile compounds produced during the process; such evaluation was performed both immediately after cooking (T0) and after 7 days (T7) of cooked-meat storage at 4 °C. Cooking-loss values were higher for samples from chickens raised with Neutral LED (p < 0.05) compared to the other groups. For the fatty acid profiles of the meat, higher values were found for monounsaturated fatty acids (MUFAs) such as C18:1, C9 and C16:1 in Cool LED compared to the Control. Regarding the volatile profile of cooked meat, compounds belonging to the families of aldehydes, alcohols, ketones, and aromatic compounds were identified. Compounds belonging to the aldehyde family, such as hexanal, increased in Cool LED samples at T0 in comparison to the Control. On the other hand, the amounts of 1-Pentanol, 1-Octanol and 2-Octen-1-ol, which belong to the alcohol family, increased at T7 in Cool LED samples compared to the Warm LED. In conclusion, LED lighting showed to be effective in inducing significant variations on chicken breast meat ready to be introduced to the market, in particular regarding fatty acid profiles and the accumulation of volatile compounds. However more in-depth evaluation is needed for the identification of modifications regarding the sensorial sphere, which could have an impact on the consumer acceptability of the product

Whole Blood Transcriptome Profiling Reveals Positive Effects of Olive Leaves-Supplemented Diet on Cholesterol in Goats

Agro-industrial by-products represent an important source of compounds credited with high biotechnological potential. In the last decade, considerable interest has developed toward the use of these matrices as dietary supplements in the zootechnical field, paying particular attention to the qualitative aspects associated with animal products. However, less is known about the effect of these matrices on gene expression and thus on animal metabolism. Therefore, the aim of this study was to analyze the whole blood transcriptome of lactating goats fed a dietary supplementation with 10% olive leaves (OL), one of the main by-products deriving from the olive oil chain supply. By applying a false discovery rate (FDR) &lt; 0.05 and a Log2 Fold change (Log2Fc) lower than −0.5 or higher than +0.5, it was possible to identify the differential regulation of gene coding for the apolipoprotein B (apoB) mRNA editing enzyme catalytic subunit 2 (APOBEC2), which showed downregulation in goats that received the dietary supplementation. An evaluation of both blood and milk cholesterol was performed, taking into account the strong association between plasma apoB and low-density lipoprotein (LDL). Results showed significantly lower concentrations of circulating cholesterol and cholesterol released into the milk through the mammary gland, demonstrating positive effects of OL feeding on animal welfare and potential health benefits for consumers

Intravenous neridronate in adults with osteogenesis imperfecta

Intravenous neridronate in adults with osteogenesis imperfect

Pattern of fractures across pediatric age groups: analysis of individual and lifestyle factors

Abstract Background Knowledge of the epidemiology of children's fractures is essential to develop preventive strategies. The aim of this study was to analyze the individual/lifestyle determinants of fractures across pediatric age groups. Methods A cross-sectional study was performed in the first six months of 2008 through questionnaire on a sample of children from an outpatient clinic for pediatric fractures. Differences in gender, anatomic site, circumstances and location of fracture occurrence, behavioural lifestyle, and calcium intake were investigated among three different age classes (pre-school children, school children, and adolescents). Results The sample consisted of 382 subjects (2-14 years of age) sustaining a fracture after low or moderate trauma. Males were at a higher risk of fractures than females; greater than two-thirds of injuries occurred after low-energy trauma and the upper limb was more frequently involved. With increasing age, the male/female ratio and time spent in sports participation increased (p Conclusions Gender differences were shown in the prevalence of injuries, characteristics, and circumstances across ages. These differences may be explained by the related changes in behaviors, together with attending different places. Individual and lifestyle factors can in part explain the variability in the occurrence of fractures and can also address targeted preventive strategies.</p