Expression of active and inactive forms of CIK and CARMA 2sh proteins in Normal Human Epidermal Keratinocytes (NHEK)

Psoriasis is a debilitating skin disease affecting approximately 23% of human population. The disease is considered to have key genetic underpinning and genome wide association studies and meta analysis have identified more than 40 susceptibility loci for psoriasis. CARMA proteins play a major role in regulating activation of transcription factor NF-kB which is ubiquitously expressed in mammalian cells that play a central role in the control of immune and inflammatory response. Missense mutations in the CARMA2 gene have been shown to dominantly transmit the psoriatic trait with high penetrance. Most of the CARMA2 mutations associated with skin disorders clusters in the coiled coil domain of the protein.NPRP 7-466-3-119 from the Qatar National Research Fund of Qatar Foundatio

Effect of CARMA 2sh gene in Mouse embryonic stem (ES) cells

CARMA2 belongs to the CARMA family of proteins. They are involved in the regulation and activation of NF-κB that have a central role in the control of immune and inflammatory response, and cell survival and proliferation. CARMA2short (CARMA2sh) which is the most prominent CARMA2 isoform expressed in human keratinocytes. It has already been identified that CARMA2sh induces activation of NF-κB, and this activity requires the function of another CARD-containing protein, namely BCL10, and the adapter protein TRAF2. This study identified a CARMA Inhibitory Kinase (CIK) which inhibit the ability to induce NF-κB. CIK is not tested for their function in Human Primary keratinocytes and hence we attempt to understand the function of CIK and its associated molecules by invitro and invivo models. The inhibitory activity of CIK on CARMA2 in primary human keratinocytes expressing wild (wt) & mutant CARMA2 was analyzed.This work was made possible by NPRP grant NPRP 7 - 466 - 3119 from the Qatar National Research Fund of Qatar Foundatio

Effect of psoriasis linked CARMA2sh gene in transgenic animal model

Background • CARMA2 belongs to the CARMA family of proteins. They are involved in the regulation and activation of NF-κB, that has a central role in controlling the immune and inflammatory response, cell survival and proliferation. • CARMA2short (CARMA2sh) is the most prominent CARMA2 isoform expressed in human keratinocytes • It has already been identified that CARMA2sh induces activation of NF-κB in association with another CARD-containing protein, namely BCL10, and the adapter protein TRAF2. • This study identified a CARMA Inhibitory Kinase(CIK) which inhibits the ability to induce NF-κB • CIK is not tested for their function in Human Primary keratinocytes and hence we attempt to understand the function of CIK and its associated molecules by invitro & invivo models • The inhibitory activity of CIK on CARMA2 in primary human keratinocytes expressing wild (wt) & mutant CARMA2 was analyzedNPRP grant NPRP 7 - 466 - 3119 from the Qatar National Research Fund of Qatar Foundatio

Animal Health Monitoring Program (AHMP) of LARC

Animal health monitoring is important to maintain the health of the animals used for research purpose. Maintaining healthy research animals is the prime requisite for best research outcomes. Health monitoring is performed as per the recommendation issued by the Federation of European Laboratory Animal Science Associations (FELASA)

Gut Microbiota Dysbiosis and Their Impact on Biochemical and Inflammatory Profiles in Cafeteria Diet Fed Sprague Dawley Rats

Gut microbiota plays a major role in regulating the host metabolism and immune system. However, the structure of microbiome population is altered constantly by diverse factors including diet and environment. In particular, the gut microbiome dynamics is influenced by diet composition and their associated metabolites. Many studies in the recent past reported on diet induced dysbiosis in the gut microbiome, the modulating ratio between Firmicutes and Bacteroidetes plays a central role in maintaining the microbiome diversity in the gut and their abundance regulates obese conditions. Although there are several reports on gut microbial dysbiosis (Firmicutes/Bacteroidetes) at phylum level but only few of them highlight at the genera level. In the present study, we focused more on the impact of cafeteria diet (CAF) with respect to the gut microbiome richness at the genera level in SD rats. Three weeks old Sprague Dawley (SD) rats were fed with normal chow diet and cafeteria diet (CAF). After 10 weeks, serum, tissue samples (small intestine and cecum), cecum fecal and fecal pellet were collected. Biochemical analysis from serum, Gene expression analysis of pro-inflammatory markers from tissues and microbiome analysis from fecal samples were analysed. CAF diet fed rat in the present study developed obesity with increased body weight, few of them developed the resistance to weight gain (WGR) and these animal shows significantly increased abundance of Bacteriodetes-Prevotella compared to obese animals. This study suggests that detailed research needed to address the contribution of microbiota abundance at the genera level. We further explored the influence of diet induced microbiota changes on immune response; increase in Bacteroidetes leads to increased LPS and this in turn elicit immune reaction in the gut. Moreover, variation in gut microbiota also affects the gut barrier function (Tight junction proteins) and increase metabolic endotoxemia that leads to activation of innate immune system and low-grade inflammation. Further studies on these lines targeting the microbiota changes in the gut at the genera level is warranted to gain more knowledge

Diagnostic Microbiology Laboratory Monitoring of LARC

Microbial free environment of a research facility is critical to maintain animal health. Establishing Microbiological Monitoring Program is mandatory to assure the quality of the system. Microbiological monitoring involves a set of microbial testing methods to detect, isolate and eliminate the harmful microorganisms to provide Specific Pathogen Free (SPF) environment within the facility. Microbial diagnostic practices should continuously monitor all the potential transmission routes to improve the methods and limiting the bioburden of clean areas of vivarium. Use of selective media and biochemical testing procedures can further identify the organisms to their genes & species leve

LABORATORY ANIMAL RESEARCH CENTRE (LARC) “A SPECIFIC PATHOGEN FREE RODENT FACILITY” - COVID – 19 PANDEMIC RESPONSE PLAN

Laboratory Animal Research Centre (LARC) has developed an early emergency operational plan for COVID-19 pandemic situation. Biosafety and biosecurity measures were planned and implemented ahead of time to check the functional requirement to prevent the infection. Identified necessary support for IT, Transport, Procurement, Finance, Admin and Research to make the operations remotely and successfull

Gut Microbiota Dysbiosis in Cafeteria Diet Fed Sprague Dawley Rats

Diet plays a major role in the body physiology and metabolism. The quantity, nature and stability of the macronutrients present in the diet have a major impact on the composition of gut microbiota. Gut microbiota plays a major role in the body metabolism and leads to obese or lean phenotype. Bacteriodetes, Firmicutes, Proteobacteria and Actinobacteria are the major microbes that inhabit in the region of the gut. We made an attempt to study the effects of Cafeteria (CAF) diets and normal chow diets on diet consumption, weight gain, metabolism and composition of gut microbiota in fecal and cecum samples from three weeks old Sprague Dawley (SD) rats (n = 18/group) using 16S rDNA high throughput sequencing. Results revealed that distinctive diet based phenotypical changes were observed in some of the Cafeteria diet fed rats. Interestingly, some weight gain resistant (WGR) animals in Cafeteria diet fed groups show similar trend like that of control normal chow fed rats. Fecal microbiome analysis indicates that the ratio of Bacteriodetes is higher than the Firmicutes in cecum samples of Cafeteria diet fed rats whereas no significant difference is found in fecal samples of Cafeteria diet fed rats and as well as in control rats. Further analysis of other taxa at the level of family and genus of microbial abundance are also discussed. Our study suggests that contribution of gut microbiota towards obesity is not at the phylum level, and microbiome composition even at the level of species or strain may exert impact on the metabolism of the Cafeteria diet

Early-Life Sugar Consumption Affects the Microbiome in Juvenile Mice

Scope: The composition of the gut microbiota is influenced by the dietary nutrient. Sugar has been linked with many metabolic health disorders such as heart disease, metabolic syndrome, and immune disorders. Long-term consumption of sugar influences the landscape of gut microbiota by altering the gut microbial population called dysbiosis. This study aims to evaluate the impact of long-term consumption of high sugar diet (HSD) on the diversity of gut microbiota. Methods and results: CD1 mice are given high concentration of sugar for 15 weeks followed by a recovery period of 10 weeks. Real-time polymerase chain reaction and 16S rRNA next-generation sequencing methods employ to identify microbiome diversity. The results show that Firmicutes and Bacteroidetes are the predominant phyla in control, cecum, and fecal samples. Firmicutes population are gradually increased in treated samples even after the recovery period, whereas Bacteroidetes abundance slightly reduces throughout the study. Conclusion: The present study shows that the impact of long period of high sugar diet consumption alters the diversity of normal gut flora which can be restored after 10 weeks of sugar withdrawal. This indicates that the intervention of healthy and nutritious diet influences gut microbes and this can be beneficial in reducing the implication of early life metabolic disorders such as obesity.The authors would like to acknowledge LARC for their support and LARC vivarium team, for their support in animal care and husbandry. The research is funded by Qatar University