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Metabolic labeling is often used to measure protein turnover. Here the authors show that for interconvertible protein species like phosphoforms metabolic labeling does not provide information on turnover differences, but that the relative order of modification can determine the observed dynamics.

Highly concentrated antibody formulations are oftentimes required for subcutaneous, self-administered biologics. Here, we report the development of a unique formulation for our first-in-class FSH-blocking humanized antibody, MS-Hu6, which we propose to move to the clinic for osteoporosis, obesity, and Alzheimer’s disease. The studies were carried out using our Good Laboratory Practice (GLP) platform, compliant with the Code of Federal Regulations (Title 21, Part 58). We first used protein thermal shift, size exclusion chromatography, and dynamic light scattering to examine MS-Hu6 concentrations between 1 and 100 mg/mL. We found that thermal, monomeric, and colloidal stability of formulated MS-Hu6 was maintained at a concentration of 100 mg/mL. The addition of the antioxidant L-methionine and chelating agent disodium EDTA improved the formulation’s long-term colloidal and thermal stability. Thermal stability was further confirmed by Nano differential scanning calorimetry (DSC). Physiochemical properties of formulated MS-Hu6, including viscosity, turbidity, and clarity, confirmed with acceptable industry standards. That the structural integrity of MS-Hu6 in formulation was maintained was proven through Circular Dichroism (CD) and Fourier Transform Infrared (FTIR) Spectroscopy. Three rapid freeze–thaw cycles at –80 °C/25 °C or –80 °C/37 °C further revealed excellent thermal and colloidal stability. Furthermore, formulated MS-Hu6, particularly its Fab domain, displayed thermal and monomeric storage stability for more than 90 days at 4°C and 25°C. Finally, the unfolding temperature (Tm) for formulated MS-Hu6 increased by >4.80 °C upon binding to recombinant FSH, indicating highly specific ligand binding. Overall, we document the feasibility of developing a stable, manufacturable and transportable MS-Hu6 formulation at a ultra-high concentration at industry standards. The study should become a resource for developing biologic formulations in academic medical centers.

Abstract The aim of this study was the isolation and molecular characterization of fungi from untreated refinery effluent by using multiple conserved genes. The Fungi isolated were characterized based on PCR amplification and genomic sequencing of the internal transcribed spacer region (ITS), partial β-tubulin (BenA), calmodulin (CaM), and RNA polymerase second large subunit (RPB2) genes, along with morphological characterization. The obtained sequences were subjected to BLAST analysis and the corresponding fungal isolates were assigned species names after comparison with representative sequences available in GenBank. Fifteen (15) Fungi species belonging to four genera of Aspergillus, Penicillium, Fusarium, and Trichoderma with Aspergillus as the predominant genus were identified. Therefore these genes should be used as molecular markers for species level identification of fungi (especially Aspergillus and Penicillium as proven in this study.

BackgroundSimultaneous presence of elevated waist circumference and hypertriglyceridemia (HTGW) is a simple and low-cost measure of visceral obesity, and it is associated with a plethora of cardio-metabolic abnormalities that can increase the risk of cardiovascular diseases and incident Type 2 diabetes mellitus. We decided to study the prevalence, patterns, and predictors of metabolic abnormalities in Nigerian hypertensives with the HTGW phenotype.MethodsThe medical records of 582 hypertensives with complete data of interest were retrieved and analyzed for the study. Their socio-demographic data, anthropometric data, and booking blood pressure values were retrieved. The results of their fasting plasma glucose, lipid profile, uric acid and serum creatinine were also retrieved for analysis.ResultsThe mean age of the study population was 56.2 ±13.6, with 53.1% being males. The prevalence of smoking and use of alcohol was 4.3% and 26.5% respectively. The prevalence of the HTGW phenotype was 23.4% and were predominantly males (61%). Subjects with the HTGW phenotype were more obese assessed by waist circumference (WC) and body mass index (BMI). Mean serum total cholesterol, triglyceride, very low-density lipoprotein, uric acid, and creatinine were significantly higher in the HTGW phenotype (p = 0.003; ConclusionThe HTGW phenotype is common amongst Nigerian hypertensives, and it is associated with metabolic abnormalities.

Peripheral arterial disease (PAD) is the least studied complication of nephrotic syndrome (NS). Risk factors which predispose children with NS to developing PAD include hyperlipidaemia, hypertension and prolonged use of steroids. The development of PAD significantly increases the morbidity and mortality associated with NS as such children are prone to sudden cardiac death. The ankle brachial index (ABI) is a tool that has been proven to have high specificity and sensitivity in detecting PAD even in asymptomatic individuals. We aimed to determine the prevalence of PAD in children with NS and to identify risk factors that can independently predict its development. A comparative cross-sectional study was conducted involving 200 subjects (100 with NS and 100 apparently healthy comparative subjects that were matched for age, sex and socioeconomic class). Systolic blood pressures were measured in all limbs using the pocket Doppler machine (Norton Doppler scan machine). ABI was calculated as a ratio of ankle to arm systolic blood pressure. PAD was defined as ABI less than 0.9. The prevalence of PAD was significantly higher in children with NS than matched comparison group (44.0% vs 6.0%, p < 0.001). Average values of waist and hip circumference were significantly higher in subjects with PAD than those without PAD (61.68± 9.1cm and 67.6± 11.2 cm vs 57.03 ± 8.3cm and 65.60± 12.5cm respectively, p< 0.005). Serum lipids (triglyceride, very low density lipoprotein, total cholesterol and low density lipoprotein) were also significantly higher in subjects with PAD than those without PAD [106.65mg/dl (67.8-136.7) vs 45.72mg/dl (37.7-61.3), 21.33mg/dl (13.6-27.3) vs 9.14mg/dl (7.5-12.3), 164.43mg/dl (136.1-259.6) vs 120.72mg/dl (111.1-142.1) and 93.29mg/dl (63.5-157.3) vs 61.84mg/dl (32.6-83.1), respectively p< 0.05]. Increasing duration since diagnosis of NS, having a steroid resistant NS and increasing cumulative steroid dose were independent predictors of PAD in children with NS; p< 0.05 respectively. With these findings, it is recommended that screening for PAD in children with NS should be done to prevent cardiovascular complications before they arise.

Animal whole-brain functional magnetic resonance imaging (fMRI) provides a non-invasive window into brain activity. A collection of associated methods aims to replicate observations made in humans and to identify the mechanisms underlying the distributed neuronal activity in the healthy and disordered brain. Animal fMRI studies have developed rapidly over the past years, fueled by the development of resting-state fMRI connectivity and genetically encoded neuromodulatory tools. Yet, comparisons between sites remain hampered by lack of standardization. Recently, we highlighted that mouse resting-state functional connectivity converges across centers, although large discrepancies in sensitivity and specificity remained. Here, we explore past and present trends within the animal fMRI community and highlight critical aspects in study design, data acquisition, and post-processing operations, that may affect the results and influence the comparability between studies. We also suggest practices aimed to promote the adoption of standards within the community and improve between-lab reproducibility. The implementation of standardized animal neuroimaging protocols will facilitate animal population imaging efforts as well as meta-analysis and replication studies, the gold standards in evidence-based science.

The activated mutants of the α-subunits of G proteins G12 and G13 have been designated as the gep oncogenes owing to their ability to stimulate diverse oncogenic signaling pathways that lead to neoplastic transformation of fibroblast cell lines and tumorigenesis in nude mice models. Studies from our laboratory as well as others have shown that the growth-promoting activities of Gα12 and Gα13 involve potent activation of c-Jun N-terminal kinases (JNKs). Our previous studies have indicated that the JNK-interacting leucine zipper protein (JLP), a scaffold protein involved in the structural and functional organization of the JNK/p38 mitogen-activated protein kinase module, tethers Gα12 and Gα13 to the JNK signaling module. In the present study, in addition to demonstrating the physical association between JLP and Gα12, we show that this interaction is enhanced by the receptor- or mutation-mediated activation of Gα12. We also establish that JLP interacts with Gα12 through the C-terminal domain that has been previously identified to be involved in binding to Gα13. Furthermore, using this C-terminal domain as a competitively inhibitor of JLP that can disrupt Gα12-JLP interaction, we demonstrate that JLP is required for the stimulation of JNK by Gα12. Our results also indicate that such JLP interaction is required for Gα12 as well as Gα13-mediated neoplastic transformation of JLP. These studies demonstrate for the first time a functional role for JLP in the gep oncogene-regulated neoplastic signaling pathway.

he newborn. The study team developed the train-the-trainer program which was delivered at the University of Buea in the Southwest region of Cameroon in March, 2011. TTT is particularly effective for reaching large audiences and also permits a degree of sustainability such that the Cameroonian students will be trainers for subsequent cohorts of their peers. This study continues to strengthen the collaborative endeavors between the two nursing schools; the University of Buea (UB) and Goldfarb School of Nursing (GSON) at Barnes Jewish College in Saint Louis, Missouri, USA. The final aim of the intervention was the initiation of collaborative relationships between the faculty members of the two educational organizations.

HIV/AIDS is a major public health pandemic affecting the development, survival and life of young people both in Cameroon and the USA. Youths are more adaptive to change and less hindered by prejudice than adults. Student-to-student peer mentoring is a non-formal way for students to learn new life skills and different cross-cultural values. We report on a two-day consultative meeting on developing international collaboration for the fight against HIV/AIDS between Cameroonian and USA nursing students. We used adult learning approaches consisting of presentations, discussions, questions and answer sessions, role plays and demonstrations. Deliberations and resolutions from the consultative meeting enabled the Vice-Chancellor of the University of Buea to sign a memorandum of understanding with the Goldfarb School of Nursing in the USA on HIV/AIDS international collaboration paving the way forward for more developmental health projects in this domain.

Pathogen-pattern-recognition by Toll-like receptors (TLRs) and pathogen clearance after immune complex formation via engagement with Fc receptors (FcRs) represent central mechanisms that trigger the immune and inflammatory responses. In the present study, a linkage between TLR4 and FcgammaR was evaluated in vitro and in vivo. Most strikingly, in vitro activation of phagocytes by IgG immune complexes (IgGIC) resulted in an association of TLR4 with FcgammaRIII (CD16) based on co-immunoprecipitation analyses. Neutrophils and macrophages from TLR4 mutant (mut) mice were unresponsive to either lipopolysaccharide (LPS) or IgGIC in vitro, as determined by cytokine production. This phenomenon was accompanied by the inability to phosphorylate tyrosine residues within immunoreceptor tyrosine-based activation motifs (ITAMs) of the FcRgamma-subunit. To transfer these findings in vivo, two different models of acute lung injury (ALI) induced by intratracheal administration of either LPS or IgGIC were employed. As expected, LPS-induced ALI was abolished in TLR4 mut and TLR4(-/-) mice. Unexpectedly, TLR4 mut and TLR4(-/-) mice were also resistant to development of ALI following IgGIC deposition in the lungs. In conclusion, our findings suggest that TLR4 and FcgammaRIII pathways are structurally and functionally connected at the receptor level and that TLR4 is indispensable for FcgammaRIII signaling via FcRgamma-subunit activation.

The book "Salah Ben Youssef et les youssefistes: Au tournant de l'indépendance tunisienne, 1955–1956" by M'hamed Oualdi explores the rise and fall of the Youssefist movement in the Tunisian national movement. The author focuses on Salah Ben Youssef's leadership position in the Neo-Destour party, his expulsion from the party, and the support he garnered across Tunisia. The book also delves into the violent suppression of the movement after its official ban in 1956. The author draws on French Ministry of Foreign Affairs and French military archives for information. The book offers insights into Ben Youssef's background and his vision for Tunisia as a constitutional monarchy. The author questions why Tunisia did not evolve into a multiparty state like Morocco. However, the book could have benefited from further analysis of the rivalry between Bourguiba and Ben Youssef and the perspectives of other key figures. The book provides a useful chronology of events but lacks an index. [Extracted from the article]

"Les camps de regroupement en Algérie: Une histoire des déplacements forcés (1954–1962)" by Fabien Sacriste is a meticulously researched history of the forced relocation of Algeria's rural population by the French military between 1954 and 1962. The author examines the bureaucratic infighting and difficult relationships between French military and civil authorities during this period. The book also explores the negative connotations associated with the term "camp de regroupement" and the government's response to the displacement of Algerians. The author sheds light on the relationship between military tactics and modernization efforts in Algeria, as well as the long-lasting impact of the forced relocations. [Extracted from the article]