Immunoglobulin Gamma Subclasses and Corresponding Fc Receptors in Rhesus Macaques: Genetic Characterization and Engineering of Recombinant Molecules

Rhesus macaques represent a valuable model in biomedical research and in development of vaccines and therapeutics. Due to the lack of reagents, the general properties of IgG and corresponding cellular receptors (FcγR) in this species are poorly characterized. We engineered recombinant IgGs containing each of the four rhesus macaque heavy constant region (CH) subclasses. To define FcγRs that mediate IgGs, we identified and characterized three FcγR classes, and generated recombinant cDNA constructs. cDNA IgH constructs were created by fusing – by sequence overlap extension PCRs – a gene segment encoding the murine variable heavy domain specific for the hapten NIP, an established specificity system for assessing antibody effector functions, with rhesus macaque CH fragments. The complete IgH constructs were transfected into J558L cells, a murine IgH-lost myeloma cell line expressing anti-NIP light chain. Secretion of engineered IgGs was determined by ELISAs using NIP-BSA and anti-monkey IgG-specific antibodies. Molecular cloning methods were applied to identify and clone FcγR genes, and recombinant FcγR cDNA constructs were created by the recombinant DNA method. Four engineered IgH cDNA constructs were successfully created. Recombinant IgGs, in the intact Ig form and retaining the original anti-NIP specificity, were successfully produced. Compared to those in humans, FcγRs in rhesus macaques share high homology, yet also feature a relatively high level of intra-species polymorphism and possess different N-linked glycosylation patterns. FcγR constructs and expression vectors were successfully generated. The chimeric recombinant IgGs are powerful tools for defining IgG functional properties and studying CH structure/function relationship. These molecules can also be used as immunogens for generation of antibodies capable of unequivocally detecting individual IgG subclasses. The findings on FcγRs validate rhesus macaques as a model for studying antibody responses, and underscore the need to take into account of the genetic heterogeneity. The FcγR constructs and vectors serve as a tool for further studies of IgG/FcγR interactions. We also reported here our findings from a separate study that the main female hormone, 17β-estradiol, is capable of restoring antibody responses to an influenza vaccine in a postmenopausal mouse model, suggesting that immunogenicity and efficacy of influenza vaccines should be evaluated in postmenopausal women

Interfacial Layers between Ion and Water Detected by Terahertz Spectroscopy

Dynamic fluctuations in hydrogen-bond network of water occur from femto- to nano-second timescale and provides insights into structural/dynamical aspects of water at ion-water interfaces. Employing terahertz spectroscopy assisted with molecular dynamics simulations, we study aqueous chloride solutions of five monovalent cations, namely, Li, Na, K, Rb and Cs. We show that ions modify the behavior of surrounding water molecules and form interfacial layers of water around them with physical properties distinct from that of bulk water. Small cations with high charge densities influence the kinetics of water well beyond the first solvation shell. At terahertz frequencies, we observe an emergence of fast relaxation processes of water with their magnitude following the ionic order Cs>Rb>K>Na>Li, revealing an enhanced population density of weakly coordinated water at ion-water interface. The results shed light on the structure breaking tendency of monovalent cations and provide insights into the properties of ionic solutions at the molecular level

Effects of salinity and alkalinity on growth and survival of all-male giant freshwater prawn (Macrobrachium rosenbergii De Man, 1879) juveniles

All-male giant freshwater prawns (AMGFPs) have been a popular crop cultivated in the Mekong Delta, Vietnam, due to their proven production efficiency compared to all-female or mixed-sex prawn cultures. However, the crucial water quality factors impacting AMGFP aquaculture efficiency have yet to be elaborately investigated. Two separate experiments were randomly arranged with three replicates to evaluate the effects of salinity or alkalinity on the growth and survival of AMGFP juveniles during the grow-out period. The results show that the prawn survival rate in the salinity range of 0–15‰ varied from 66.1 to 74.8％ and in a salinity range of 0–5‰ was relatively low compared to the range of 10-15‰; however, the difference was not significant among salinities after 90 days of culture (p > 0.05). All the prawn growth performance parameters significantly decreased with increasing salinities of 0, 5, 10, and 15‰ after 30, 60, and 90 days of culture (p 0.05), and both were significantly higher than those at salinities of 10 and 15‰ (p < 0.05) after 90 days of culture. In addition, the survival rate reached 82.5–84.4％ and did not significantly differ among alkalinities of 80, 100, 120, 140, and 160 mgCaCO3 L−1. However, the growth performance parameters and yield of AMGFPs at an alkalinity of 160 mg L−1 were significantly higher than those at lower alkalinities (80, 100, 120, and 140 mg CaCO3 L−1) after 90 days of culture. Therefore, it is recommended that a salinity range of 0–5‰ and alkalinity of 160 mgCaCO3 L−1 is optimal for the growth-out culture of AMGFP juveniles

The Emergence of Rotavirus G12 and the Prevalence of Enteric Viruses in Hospitalized Pediatric Diarrheal Patients in Southern Vietnam

Diarrhea is a major cause of childhood morbidity and mortality in developing countries, and the majority of infections are of viral etiology. We aimed to compare the etiological prevalence of the major enteric viruses in an urban and a rural setting in southern Vietnam. We simultaneously screened fecal specimens from 362 children in Ho Chi Minh City and Dong Thap province that were hospitalized with acute diarrhea over a 1-month-long period for four viral gastrointestinal pathogens. Rotavirus was the most common pathogen identified, but there was a differential prevalence of rotavirus and norovirus between the urban and rural locations. Furthermore, rotavirus genotyping and phylogenetic analysis again differentiated the genotypes by the sampling location. Our data show a disproportional distribution of enteric viral pathogens in urban and rural locations, and we provide evidence of continual importation of new rotavirus strains into southern Vietnam and report the emergence of rotavirus genotype G12

Habitat Delineation in Highly Variable Marine Environments

The structure of the phytoplankton community in surface waters is the consequence of complex interactions between the physical and chemical properties of the upper water column as well as the interaction within the general biological community. Understanding the structure of phytoplankton communities is especially challenging in highly variable and dynamic marine environments. A variety of strategies have been employed to delineate marine planktonic habitats, including both biogeochemical and water-mass-based approaches. These methods have led to fundamental improvements in our understanding of marine phytoplankton distributions, but they are often difficult to apply to systems with physical and chemical properties and forcings that vary greatly over relatively short spatial or temporal scales. In this study, we have developed a method of dynamic habitat delineation based on environmental variables that are biologically relevant, that integrate over varying time scales, and that are derived from standard oceanographic measurements. As a result, this approach is widely applicable, simple to implement, and effective in resolving the spatial distribution of phytoplankton communities. As a test of our approach, we have applied it to the Amazon River-influenced Western Tropical North Atlantic (WTNA) and to the South China Sea (SCS), which is influenced by both the Mekong River and seasonal coastal upwelling. These two systems differ substantially in their spatial and temporal scales, nutrient sources/sinks, and hydrographic complexity, providing an effective test of the applicability of our analysis. Despite their significant differences in scale and character, our approach generated statistically robust habitat classifications that were clearly relevant to surface phytoplankton communities. Additional analysis of the habitat-defining variables themselves can provide insight into the processes acting to shape phytoplankton communities in each habitat. Finally, by demonstrating the biological relevance of the generated habitats, we gain insights into the conditions promoting the growth of distinct communities and the factors that lead to mismatches between environmental conditions and phytoplankton community structure

MyCTC chip: microfluidic-based drug screen with patient-derived tumour cells from liquid biopsies

Cancer patients with advanced disease are characterized by intrinsic challenges in predicting drug response patterns, often leading to ineffective treatment. Current clinical practice for treatment decision-making is commonly based on primary or secondary tumour biopsies, yet when disease progression accelerates, tissue biopsies are not performed on a regular basis. It is in this context that liquid biopsies may offer a unique window to uncover key vulnerabilities, providing valuable information about previously underappreciated treatment opportunities. Here, we present MyCTC chip, a novel microfluidic device enabling the isolation, culture and drug susceptibility testing of cancer cells derived from liquid biopsies. Cancer cell capture is achieved through a label-free, antigen-agnostic enrichment method, and it is followed by cultivation in dedicated conditions, allowing on-chip expansion of captured cells. Upon growth, cancer cells are then transferred to drug screen chambers located within the same device, where multiple compounds can be tested simultaneously. We demonstrate MyCTC chip performance by means of spike-in experiments with patient-derived breast circulating tumour cells, enabling >95% capture rates, as well as prospective processing of blood from breast cancer patients and ascites fluid from patients with ovarian, tubal and endometrial cancer, where sensitivity to specific chemotherapeutic agents was identified. Together, we provide evidence that MyCTC chip may be used to identify personalized drug response patterns in patients with advanced metastatic disease and with limited treatment opportunities

Formation and Degradation of Beta-casomorphins in Dairy Processing

Milk proteins including casein are sources of peptides with bioactivity. One of these peptides is beta-casomorphin (BCM) which belongs to a group of opioid peptides formed from b-casein variants. Beta-casomorphin 7 (BCM7) has been demonstrated to be enzymatically released from the A1 or B b-casein variant. Epidemiological evidence suggests the peptide BCM 7 is a risk factor for development of human diseases, including increased risk of type 1 diabetes and cardiovascular diseases but this has not been thoroughly substantiated by research studies. High performance liquid chromatography coupled to UV-Vis and mass spectrometry detection as well as enzyme–linked immunosorbent assay (ELISA) has been used to analyze BCMs in dairy products. BCMs have been detected in raw cow’s milk and human milk and a variety of commercial cheeses, but their presence has yet to be confirmed in commercial yoghurts. The finding that BCMs are present in cheese suggests they could also form in yoghurt, but be degraded during yoghurt processing. Whether BCMs do form in yoghurt and the amount of BCM forming or degrading at different processing steps needs further investigation and possibly will depend on the heat treatment and fermentation process used, but it remains an intriguing unknown