Small Changes Huge Impact: The Role of Protein Posttranslational Modifications in Cellular Homeostasis and Disease

Posttranslational modifications (PTMs) modulate protein function in most eukaryotes and have a ubiquitous role in diverse range of cellular functions. Identification, characterization, and mapping of these modifications to specific amino acid residues on proteins are critical towards understanding their functional significance in a biological context. The interpretation of proteome data obtained from the high-throughput methods cannot be deciphered unambiguously without a priori knowledge of protein modifications. An in-depth understanding of protein PTMs is important not only for gaining a perception of a wide array of cellular functions but also towards developing drug therapies for many life-threatening diseases like cancer and neurodegenerative disorders. Many of the protein modifications like ubiquitination play a decisive role in various drug response(s) and eventually in disease prognosis. Thus, many commonly observed PTMs are routinely tracked as disease markers while many others are used as molecular targets for developing target-specific therapies. In this paper, we summarize some of the major, well-studied protein alterations and highlight their importance in various chronic diseases and normal development. In addition, other promising minor modifications such as SUMOylation, observed to impact cellular dynamics as well as disease pathology, are mentioned briefly

Bilateral facial nerve palsy: a rare complication of dengue fever – a case report

Dengue is the most common mosquito-borne viral infection worldwide. There is increased evidence for dengue virus neurotropism, and neurological manifestations could make part of the clinical picture of dengue virus infection. Pathogenic mechanisms include systemic complications and metabolic disturbances resulting in encephalopathy, direct effect of the virus provoking encephalitis, and post infectious immune mechanisms causing immune-mediated syndromes. Dengue viruses should be considered as a cause of neurological disorders in endemic regions. Standardized case definitions for specific neurological complications are still needed. We report a patient with dengue fever who later developed bilateral facial nerve palsy

A Novel Reporter Mouse Uncovers Endogenous Brn3b Expression.

Brn3b (Pou4f2) is a class-4 POU domain transcription factor known to play central roles in the development of different neuronal populations of the Central Nervous System, including retinal ganglion cells (RGCs), the neurons that connect the retina with the visual centers of the brain. Here, we have used CRISPR-based genetic engineering to generate a Brn3b-mCherry reporter mouse without altering the endogenous expression of Brn3b. In our mouse line, mCherry faithfully recapitulates normal Brn3b expression in the retina, the optic tracts, the midbrain tectum, and the trigeminal ganglia. The high sensitivity of mCherry also revealed novel expression of Brn3b in the neuroectodermal cells of the optic stalk during early stages of eye development. Importantly, the fluorescent intensity of Brn3b-mCherry in our reporter mice allows for noninvasive live imaging of RGCs using Scanning Laser Ophthalmoscopy (SLO), providing a novel tool for longitudinal monitoring of RGCs

Characterization of sperm heparin binding proteins (HBPs) using polyclonal antibodies raised against seminal plasma HBPs: Application in buffalo bull fertility

26-33This study aimed to evaluate rabbit polyclonal antibodies raised against purified seminal plasma sperm membrance extracts (SP) heparin binding protein (HBP) for identifying HBPs in buffalo bull spermatozoa by western blotting. Anti-SP-HBP recognized 11 polypeptides in SDS-sperm membrance extracts (SME) of 31 tested bulls. Thirty one bulls were divided into G-1 (>40%) and G-II (≤40%) based on acrosome reaction. Immunoblotting revealed that HBPs of 24, 30, 38 and 43 kDa were present in 3%, 7.02%, 1.16% and 4.83% more bulls of G-I, whereas, 20 and 46 kDa HBPs were present in 13.2 and 9.65% more bulls of G-II. Immunoblotting of anti-HBP with sperm extracts of 10 bulls (22-31) indicated that 31 kDa positive bulls had 10.9% higher conception rate than 31 kDa negative bulls. Although 24 kDa HBP was detected in 10 bulls, but its expression was very weak in bull number 22, 23 and 26, which had 10.7% lower conception rate than the bulls with strong expression of 24 kDa HBP. In the present study, 17/20 kDa positive bulls exhibited 4.46% and 8.67% low conception rate than 17/20 kDa negative bulls. Mass spectrometry analysis revealed matching of 24, 31, 33 and 38 kDa proteins with MHC class 1 antigen, tRNA methyl transferase 11 homolog partial, parvalbumin alpha-like and cilia- and flagella-associated protein 99. This study suggests that buffalo bull fertility can be predicted from sperm HBP

Manganese provides antioxidant protection for sperm cryopreservation that may offer new consideration for clinical fertility

Reactive oxygen species (ROS) are generated by sperm metabolism. While, ROS are required for maturation, capacitation and acrosome reaction, they also modify many peroxidable cellular compounds. There is production of ROS during cryopreservation and frozen spermatozoa are highly sensitive to lipid peroxidation (LPO). Antioxidants exert a protective effect on the plasma membrane of frozen bovine sperm preserving both metabolic activity and cellular viability. Manganese (Mn++) is proved to be a chain breaking antioxidant in biological system. Therefore, we examined the role of (Mn++) during cryopreservation of cattle bull semen. Semen was divided into four parts and cryopreserved in egg-yolk-citrate extender + glycerol (EYC-G), EYC-G + 100 µM of Mn++, EYC-G + 150 µM of Mn++ and EYC-G + 200 µM of Mn++. After four hours of cooling and 24 hrs of freezing, the spermatozoa were examined for percentage motility, Hypo-osmotic swelling (HOS), LPO and protein leakage. Addition of manganese to the semen during cryopreservation showed a protective effect and accounted for an increase in semen quality parameters [percentage motility, HOS percent and decrease in malondialdehyde (MDA) production and protein leakage]. The effect of manganese on motility and HOS was non-significant (p < 0.05) in cooled spermatozoa but significant with 150 µM of Mn++ in frozen-thawed spermatozoa. MDA production and protein leakage decreased to a significant and maximum level (p < 0.05) on addition of 200 µM of manganese. The addition of manganese to EYC-G dilutor will improve the quality/fertility of semen, which will result in improvement of in vitro fertilization and artificial insemination success rate

Peroxisome Proliferator-Activated Receptor-γ Coactivator 1-α (PPARGC1A) Genetic Associations with Type 2 Diabetes in Three Ethnicities

Genetic heterogeneity, lifestyle factors, gene-gene or gene-environment interactions are the determinants of T2D which puts Hispanics and populations with African ancestry at higher risk of developing T2D. In this dissertation, the genetic associations of PPARGC1A polymorphisms with T2D and its related phenotypes (metabolic markers) in Haitian Americans (cases=110, controls=116), African Americans (cases=120, controls=124) and Cuban Americans (cases=160, controls=181) of South Florida were explored. Five single nucleotide polymorphisms of gene PPARGC1A were evaluated in each ethnicity for their disease association. In Haitian Americans, rs7656250 (OR= 0.22, pp=0.03) had significant protective association with T2D but had risk association in African Americans for rs7656250 (OR=1.02, p=0.96) and rs4235308 (OR=2.53, p=0.03). We found that in Haitian American females, both rs7656250 (OR=0.23, pp=0.03) had protective association with T2D. In African American females, rs7656250 (OR=1.14, p=0.78) had risk association whereas in males, it had significant protective effect (OR=0.37, p=0.04). However, the risk association exhibited by rs4235308 was stronger in African American females (OR=2.69, p=0.03) than males (OR=1.16, p=0.72). In Cuban Americans, only rs7656250 showed significant risk association with T2D (OR=6.87, p=0.02) which was stronger in females alone (OR=7.67, p=0.01). We also observed significant differences among correlations of PPARGC1A SNPs and T2D phenotypes. Positive correlation was observed for log Hs-CRP with rs3774907 (pp=0.03) in Cuban Americans respectively. Correlation of log A1C with rs7656250 (p=0.02) was positive in Cuban Americans while it was negative for rs3774907 in Haitian Americans (ppPPARGC1A correlations with T2D and its phenotypes among the three ethnicities studied (ii) the associations of PPARGC1A SNPs showed significant effect modification by sex. The findings suggest that variations in effects of PPARGC1A gene polymorphisms among three ethnicities and between sexes may have biomedical implications for the development of T2D as well as the phenotypes related to T2D

Evaluation of a multidisciplinary Tier 3 weight management service for adults with morbid obesity, or obesity and comorbidities, based in primary care

A multidisciplinary Tier 3 weight management service in primary care recruited patients with a body mass index ≥40 kg·m−2, or 30 kg·m−2 with obesity-related co-morbidity to a 1-year programme. A cohort of 230 participants was recruited and evaluated using the National Obesity Observatory Standard Evaluation Framework. The primary outcome was weight loss of at least 5% of baseline weight at 12 months. Diet was assessed using the two-item food frequency questionnaire, activity using the General Practice Physical Activity questionnaire and quality of life using the EuroQol-5D-5L questionnaire. A focus group explored the participants' experiences. Baseline mean weight was 124.4 kg and mean body mass index was 44.1 kg·m−2. A total of 102 participants achieved 5% weight loss at 12 months. The mean weight loss was 10.2 kg among the 117 participants who completed the 12-month programme. Baseline observation carried forward analysis gave a mean weight loss of 5.9 kg at 12 months. Fruit and vegetable intake, activity level and quality of life all improved. The dropout rate was 14.3% at 6 months and 45.1% at 1 year. Focus group participants described high levels of satisfaction. It was possible to deliver a Tier 3 weight management service for obese patients with complex co-morbidity in a primary care setting with a full multidisciplinary team, which obtained good health outcomes compared with existing services

G × E Interaction and Heterosis in Elite Tomato Hybrids for Growth, Earliness and Fruit Parameters in Diverse Agro-Climatic Zones of Punjab

Six promising tomato hybrids selected from a pool of 60 F1 hybrids were evaluated for seven traits, along with the check hybrid (TH-1) at two locations falling under different agro-climatic zones of Punjab, India. G × E interaction was significant for early yield, fruit weight and total fruit yield, whereas, it was non-significant for fruit number, locule number, pericarp thickness and vine length. Overall higher mean-early-yield, fruit number, fruit weight and total yield at Ludhiana, rather than at Bathinda, may be due to higher organic carbon, available phosphorus and available potash and low electrical conductivity of the experimental soil at Ludhiana. Pooled analysis showed that hybrid TH-21 had the maximum early-yield (3.73 tha-1), fruit weight (72.7 g) and locule number (2.65), whereas, TH-23 had the highest fruit number per vine (53.7) and total fruit yield (51.2 tha-1). The magnitude of pooled standard heterosis was maximum for vine length (140.7%), followed by early yield (114.8%), total yield (88.3%), fruit number (49.7%), fruit weight (27.6%), pericarp thickness (16.4%) and locule number (-21.6%). On the basis of stability and superiority for fruit weight, fruit number, early and total yield, TH-21 was found to be the most promising hybrid, followed by TH-23