Anti-voltage-Gated Potassium Channel (VGKC) Antibodies and Acquired Neuromyotonia in Patients with Immune Dysregulation,Polyendocrinopathy, Enteropathy X-Lined (IPEX) Syndrome

Journal of Clinical Immunology -     

SpyA, a C3-like ADP-ribosyltransferase, contributes to virulence in a mouse subcutaneous model of Streptococcus pyogenes infection

Streptococcus pyogenes is an important human pathogen with an expansive repertoire of verified and putative virulence factors. Here we demonstrate that a mutant deficient in the production of the streptococcal ADP-ribosyltransferase SpyA generates lesions of reduced size in a subcutaneous mouse infection model. At early stages of infection, when the difference in lesion size is first established, inflamed tissue isolated from lesions of mice infected with spyA mutant bacteria has higher levels of mRNA encoding the chemokines CXCL1 and CCL2 than does tissue isolated from mice infected with wild-type bacteria. In addition, at these early times, the mRNA levels for the gene encoding the intermediate filament vimentin are higher in the mutant-infected tissue. As wound resolution progresses, mRNA levels of the gene encoding matrix metallopeptidase 2 are lower in mutant-infected tissue. Furthermore, we demonstrate that the spyA mutant is internalized more efficiently than wild-type bacteria by HeLa cells. We conclude that SpyA contributes to streptococcal pathogenesis in the mouse subcutaneous infection model. Our observations suggest that the presence of SpyA delays wound healing in this model.     

Newborn Screening for Primary Immunodeficiency Diseases: History,Current and Future Practice

The primary objective of population-based newborn screening is the early identification of asymptomatic infants with a range of severe diseases, for which effective treatment is available and where early diagnosis and intervention prevent serious sequelae. Primary immunodeficiency diseases (PID) are a heterogeneous group of inborn errors of immunity. Severe combined immunodeficiency (SCID) is one form of PID which is uniformly fatal without early, definitive therapy, and outcomes are significantly improved if infants are diagnosed and treated within the first few months of life. Screening for SCID using T cell receptor excision circle (TREC) analysis has been introduced in many countries worldwide. The utility of additional screening with kappa recombining excision circles (KREC) has also been described, enabling identification of infants with severe forms of PID manifested by T and B cell lymphopenia. Here, we review the early origins of newborn screening and the evolution of screening methodologies. We discuss current strategies employed in newborn screening programs for PID, including TREC and TREC/KREC-based screening, and consider the potential future role of protein-based assays, targeted sequencing, and next generation sequencing (NGS) technologies, including whole genome sequencing (WGS).     

DNA adduct formation of the food carcinogen 2-amino-3-methylimidazo[4, 5-f]quinoline (IQ) in liver, kidney and colo-rectum of rats

DNA adducts of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ)have been measured in the liver, kidney, and colorectum of maleFischer-344 rats given a single oral dose of IQ (20 mg/kg).The pattern and distribution of DNA adducts examined by ³²P-postlabelingwas similar in all tissues. N-(Deoxyguanosin-8-yl)-2-amino-3-methylimidazo-[4,5-f]quinoline(dG-C8-IQ) was the principal adduct identified and it accountedfor     

Hydrogels of N-isopropylacrylamide copolymers with controlled release of a model protein

Temperature- and pH-sensitive hydrogels, based on N-isopropylacrylamide (NiPAAm) and itaconic acid (IA), were synthesized by free radical crosslinking copolymerization in the presence of lipase from Candida rugosa. The samples were characterized for their sensitivity to the changes of external conditions and the ability to control the release of a hydrophilic model protein, lipase. These hydrogels were highly responsive to temperature and pH, at constant ionic strength. Parameters, such as the crosslinking degree and non-ionic/ionic (NiPAAm/IA) ratio, were found to impact the hydrogel structure, mechanical properties, morphology and swelling kinetics at different pH and temperatures. The hydrogels demonstrated protein loading efficiency as high as 95 wt%. Release studies of a hydrophilic model protein at a physiological temperature of 37 °C were performed at different pH values. High dependence of lipase release kinetics on hydrogel structure and the environmental pH was found, showing generally low release rates, lower in acidic media (pH 2.20) and higher at higher pHs (6.80). Lipase activity was retained even after treatment conditions that would provoke denaturation of the enzyme if it was not protected in the gel. The obtained hydrogels were found suitable for releasing therapeutic proteins in a controlled manner at specific sites in gastrointestinal tract.     

The polymerisation kinetics of lower dialkyl itaconates

The free radical polymerisation kinetics of diethyl- (DEI), dipropyl- (DnPI), dibutyl- (DnBI), and dihexyl itaconate (DHI) in the bulk were studied in the temperature range from 50 to 70°C. The concentration of the initiator, 2,2′-azoisobutyronitrile (AIBN), was varied between 0.02 and 0.085 mol/dm³. The rate of polymerisation (R_p), degree of polymerisation (DP), overall polymerization rate constant (K), the ratio of the propagation and termination rate constants (k_p/k_t^1/2), as well as the chain transfer constant to monomer (C_M) were determined. The values of R_p, K, and k_p/k_t^1/2 of the investigated monomers all increase with increasing size of the alkyl group in the ester substituent, whereas C_M decreases when going from the dimethyl to the dihexyl ester. The values of C_M are larger than the corresponding values for the alkyl esters of methacrylic acid.     

Host-pathogen interactions: leukocyte phagocytosis and associated sequelae

Polymorphonuclear leukocytes (PMNs) are a critical component of the human innate immune response and are the first line of defense against invading microorganisms. Phagocytosis of invading microbes induces production of reactive oxygen species (ROS) by PMNs, which facilitates bactericidal activity. In addition to eliminating microorganisms, phagocytosis also accelerates PMN apoptosis, a process critical for resolution of inflammation. Inasmuch as leukocyte phagocytosis and ROS production are key components of the innate immune response, we developed flow cytometric methods to evaluate these processes in human PMNs. In contrast to traditional microscopy-based analyses, the methods described herein provide objective and high throughput measures of host cell-pathogen interactions. Importantly, they can be adapted for use with a number of fluorometric probes, and bacterium and host cell of choice, and each is based upon a common phagocytosis assay system. We also describe methods to measure phagocytosis-induced PMN apoptosis with this assay system. These methods entail detecting surface-exposed phosphatidylserine (early apoptosis), and measuring PMN chromatin condensation and DNA fragmentation (late apoptosis). Taken together, these assays provide rapid and accurate assessment of critical PMN processes.