Overexpression of the Cdk5 inhibitory peptide in motor neurons rescue of amyotrophic lateral sclerosis phenotype in a mouse model.

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease that affects motor nerve cells in the brain and the spinal cord. Etiological mechanisms underlying the disease remain poorly understood; recent studies suggest that deregulation of p25/Cyclin-dependent kinase 5 (Cdk5) activity leads to the hyperphosphorylation of Tau and neurofilament (NF) proteins in ALS transgenic mouse model (SOD1G37R). A Cdk5 involvement in motor neuron degeneration is supported by analysis of three SOD1G37R mouse lines exhibiting perikaryal inclusions of NF proteins and hyperphosphorylation of Tau. Here, we tested the hypothesis that inhibition of Cdk5/p25 hyperactivation in vivo is a neuroprotective factor during ALS pathogenesis by crossing the new transgenic mouse line that overexpresses Cdk5 inhibitory peptide (CIP) in motor neurons with the SOD1G37R, ALS mouse model (TriTg mouse line). The overexpression of CIP in the motor neurons significantly improves motor deficits, extends survival and delays pathology in brain and spinal cord of TriTg mice. In addition, overexpression of CIP in motor neurons significantly delays neuroinflammatory responses in TriTg mouse. Taken together, these data suggest that CIP may serve as a novel therapeutic agent for the treatment of neurodegenerative diseases.

A truncated peptide from p35, a Cdk5 activator, prevents Alzheimer's disease phenotypes in model mice.

Alzheimer's disease (AD), one of the leading neurodegenerative disorders of older adults, which causes major socioeconomic burdens globally, lacks effective therapeutics without significant side effects. Besides the hallmark pathology of amyloid plaques and neurofibrillary tangles (NFTs), it has been reported that cyclin-dependent kinase 5 (Cdk5), a critical neuronal kinase, is hyperactivated in AD brains and is, in part, responsible for the above pathology. Here we show that a modified truncated 24-aa peptide (TFP5), derived from the Cdk5 activator p35, penetrates the blood-brain barrier after intraperitoneal injections, inhibits abnormal Cdk5 hyperactivity, and significantly rescues AD pathology (up to 70-80%) in 5XFAD AD model mice. The mutant mice, injected with TFP5 exhibit behavioral rescue, whereas no rescue was observed in mutant mice injected with either saline or scrambled peptide. However, TFP5 does not inhibit cell cycle Cdks or normal Cdk5/p35 activity, and thereby has no toxic side effects (even at 200 mg/kg), a common problem in most current therapeutics for AD. In addition, treated mice displayed decreased inflammation, amyloid plaques, NFTs, cell death, and an extended life by 2 mo. These results suggest TFP5 as a potential therapeutic, toxicity-free candidate for AD.

Topographic regulation of neuronal intermediate filaments by phosphorylation, role of peptidyl-prolyl isomerase 1: significance in neurodegeneration.

The neuronal cytoskeleton is tightly regulated by phosphorylation and dephosphorylation reactions mediated by numerous associated kinases, phosphatases and their regulators. Defects in the relative kinase and phosphatase activities and/or deregulation of compartment-specific phosphorylation result in neurodegenerative disorders. The largest family of cytoskeletal proteins in mammalian cells is the superfamily of intermediate filaments (IFs). The neurofilament (NF) proteins are the major IFs. Aggregated forms of hyperphosphorylated tau and phosphorylated NFs are found in pathological cell body accumulations in the central nervous system of patients suffering from Alzheimer's disease, Parkinson's disease, and Amyotrophic Lateral Sclerosis. The precise mechanisms for this compartment-specific phosphorylation of cytoskeletal proteins are not completely understood. In this review, we focus on the mechanisms of neurofilament phosphorylation in normal physiology and neurodegenerative diseases. We also address the recent breakthroughs in our understanding the role of different kinases and phosphatases involved in regulating the phosphorylation status of the NFs. In addition, special emphasis has been given to describe the role of phosphatases and Pin1 in phosphorylation of NFs.

The Economic Cost of Rising Non-communicable Diseases in India: A Systematic Literature Review of Methods and Estimates.

BACKGROUND AND OBJECTIVES: India has one of the world's highest proportions of out-of-pocket expenditure (OOPE) payments. The low share of public health expenditure coupled with the double burden of disease (communicable and non-communicable) has a direct financial impact on individual OOPE and an indirect impact in the form of decreasing life expectancy, reduced productivity, and hence a negative impact on economic growth. This systematic review aims to compare and assess the estimated economic cost of non-communicable diseases (NCDs) in India and ascertain the methods used to derive these estimates. METHODS: This paper reviews the past 12-year (2010-22) literature on the economic impact of health shocks due to NCDs. Three databases were searched for the literature: PubMed, Scopus, and Google Scholar. Thematic analysis has been performed to analyse the findings of the study. RESULTS: The OOPE was very high for NCDs. The increasing cost was high and unaffordable, pushing many people into financial distress measured by catastrophic payments and rising impoverishment. CONCLUSION: The results indicate both the direct and indirect impact of NCDs, but the indirect burden of loss of employment and productivity, despite its relevance, has been less studied in the literature. A robust economic analysis will allow an evidence-based policy decision perspective to reduce the rising burden of NCDs.

A 24-residue peptide (p5), derived from p35, the Cdk5 neuronal activator, specifically inhibits Cdk5-p25 hyperactivity and tau hyperphosphorylation.

The activity of Cdk5-p35 is tightly regulated in the developing and mature nervous system. Stress-induced cleavage of the activator p35 to p25 and a p10 N-terminal domain induces deregulated Cdk5 hyperactivity and perikaryal aggregations of hyperphosphorylated Tau and neurofilaments, pathogenic hallmarks in neurodegenerative diseases, such as Alzheimer disease and amyotrophic lateral sclerosis, respectively. Previously, we identified a 125-residue truncated fragment of p35 called CIP that effectively and specifically inhibited Cdk5-p25 activity and Tau hyperphosphorylation induced by Aß peptides in vitro, in HEK293 cells, and in neuronal cells. Although these results offer a possible therapeutic approach to those neurodegenerative diseases assumed to derive from Cdk5-p25 hyperactivity and/or Aß induced pathology, CIP is too large for successful therapeutic regimens. To identify a smaller, more effective peptide, in this study we prepared a 24-residue peptide, p5, spanning CIP residues Lys(245)-Ala(277). p5 more effectively inhibited Cdk5-p25 activity than did CIP in vitro. In neuron cells, p5 inhibited deregulated Cdk5-p25 activity but had no effect on the activity of endogenous Cdk5-p35 or on any related endogenous cyclin-dependent kinases in HEK293 cells. Specificity of p5 inhibition in cortical neurons may depend on the p10 domain in p35, which is absent in p25. Furthermore, we have demonstrated that p5 reduced Aß(1-42)-induced Tau hyperphosphorylation and apoptosis in cortical neurons. These results suggest that p5 peptide may be a unique and useful candidate for therapeutic studies of certain neurodegenerative diseases.

Metal-induced oxidative stress level in patients with fibromyalgia syndrome and its contribution to the severity of the disease: A correlational study.

BACKGROUND: Fibromyalgia syndrome (FMS) is an extra-articular rheumatological disease characterised by widespread chronic musculoskeletal pain. Metal-induced oxidative stress contributes to the severity of FMS. AIMS: First, this study evaluated the association between plasma levels of toxic heavy metals and essential metals with oxidative stress (OS) markers. Second, the OS markers and metal contents were correlated with the disease severity by assessing the Fibromyalgia Impact Questioner Revised (FIQR) and tender points (TP). METHOD: A total of 105 FMS patients and 105 healthy controls of similar age and sex were recruited. OS parameter such as lipid peroxidation (LPO), protein carbonyl group (PCG), nitric oxide (NO) and essential metals such as zinc (Zn), magnesium (Mg), manganese (Mn), copper (Cu) and toxic heavy metals such as aluminium (Al), arsenic (As), lead (Pb) were estimated. RESULTS: Levels of LPO, PCG, NO (p< 0.001) and Cu, Mn, and Al (p< 0.001), were significantly higher, and Mg (p< 0.001) and Zn (p< 0.001) were significantly lower in patients compared to controls. A positive association was observed between OS parameters, FIQR and TP with Cu, Al and Mn. A significant negative association was observed between Zn and Mg with FIQR, TP and OS parameters. CONCLUSION: Heavy metals such as Al induce OS parameters and decrease the levels of essential trace elements such as Mg and Zn, which may be responsible for the severity of FMS.

Nitric oxide, lipid peroxidation products, and antioxidants in primary fibromyalgia and correlation with disease severity.

BACKGROUND: Fibromyalgia syndrome (FMS) is characterized by altered pain perception with chronic, widespread musculoskeletal pain. The relationship between nitric oxide, oxidative stress and the severity of FMS has not been studied. This study evaluated NO levels in plasma, LPO products and antioxidants in Red Cell lysate in patients of FMS and correlated it with disease severity. METHODS: 105 FMS patients who fulfilled 1990 ACR Criteria and 105 age- and sex-matched healthy controls were recruited over two years from 2013 to 2015. Antioxidative enzyme activity was assessed by the estimation of catalase, glutathione peroxidase (GPx) and glutathione reductase (GR) and superoxide dismutase (SOD). Nitric oxide in plasma, MDA marker of lipid per - oxidation (LPO) in the lysate was donen for estimating oxidative stress. FIQR was used to assess the severity of fibromyalgia. RESULTS: The catalase, superoxide dismutase, glutathione reductase and glutathione peroxidase levels were significantly low in patients than controls (p<0.001). Plasma NO levels and LPO were also significantly high (p<0.05). NO and LPO levels showed a significant positive correlation with FIQR (r: 0.57, 0.8 and p: <0.001) whereas a negative correlation was observed between antioxidants (Cat, GR and GPx, but not SOD) and FIQR. CONCLUSIONS: Low antioxidants and raised LPO in RBC lysate in patients with FM together with high plasma NO correlated with the severity of FMS.

Validation, residue analysis, and risk assessment of fipronil and flonicamid in cotton (Gossypium sp.) samples and soil.

Cotton crop is highly susceptible to attack by sucking pests. Being an important oilseed and feed crop, it is essential to monitor the pesticides and ensure health protection at consumer level. Therefore, a method was validated to estimate fipronil and flonicamid in various cotton samples and risk assessment was performed. Contamination of oil in the extracts from the various oil seeds and cake samples is a major problem as this oil contaminates the column and interferes with the detection of pesticides. The present manuscript for the first time describes successful analysis of the pesticides from various cotton samples including cotton oil, seed, and cake. Quick, easy, cheap, effective, rugged, and safe (QuEChERS)-based methods were validated for estimation of fipronil and flonicamid in cotton samples and in soil by LC-MS/MS. Recoveries were within the acceptable range of 70-120% with relative standard deviation ≤ 20% and HorRat values < 0.3-1.3. R2 was > 0.99. Matrix effects of 150 and 13.5% were observed for fipronil and flonicamid, respectively, in cotton leaves. Limits of quantitation (LOQs) were in the range of 0.0004 to 0.004 mg kg-1 for fipronil and flonicamid. Cotton samples collected from a field study at different locations were analyzed. Half-life ranged from 2.2 to 5.8 for fipronil and 4.6 to 7.0 days for flonicamid. A pre-harvest interval of 33 days is suggested. The risk assessment studies at maximum residue level values showed HQ < 1 at pre-harvest interval (PHI). The methods being short and easy can be extended to estimate more types of pesticides in different oilseeds. Following a PHI of 33 days, fipronil and flonicamid can be used on cotton at standard dose. As the levels of fipronil and flonicamid were below determination limit in all the soils, the environmental risk is negligible.

TFP5, a Peptide Inhibitor of Aberrant and Hyperactive Cdk5/p25, Attenuates Pathological Phenotypes and Restores Synaptic Function in CK-p25Tg Mice.

It has been reported that cyclin-dependent kinase 5 (cdk5), a critical neuronal kinase, is hyperactivated in Alzheimer's disease (AD) and may be, in part, responsible for the hallmark pathology of amyloid plaques and neurofibrillary tangles (NFTs). It has been proposed by several laboratories that hyperactive cdk5 results from the overexpression of p25 (a truncated fragment of p35, the normal cdk5 regulator), which, when complexed to cdk5, induces hyperactivity, hyperphosphorylated tau/NFTs, amyloid-ß plaques, and neuronal death. It has previously been shown that intraperitoneal (i.p.) injections of a modified truncated 24-aa peptide (TFP5), derived from the cdk5 activator p35, penetrated the blood-brain barrier and significantly rescued AD-like pathology in 5XFAD model mice. The principal pathology in the 5XFAD mutant, however, is extensive amyloid plaques; hence, as a proof of concept, we believe it is essential to demonstrate the peptide's efficacy in a mouse model expressing high levels of p25, such as the inducible CK-p25Tg model mouse that overexpresses p25 in CamKII positive neurons. Using a modified TFP5 treatment, here we show that peptide i.p. injections in these mice decrease cdk5 hyperactivity, tau, neurofilament-M/H hyperphosphorylation, and restore synaptic function and behavior (i.e., spatial working memory, motor deficit using Rota-rod). It is noteworthy that TFP5 does not inhibit endogenous cdk5/p35 activity, nor other cdks in vivo suggesting it might have no toxic side effects, and may serve as an excellent therapeutic candidate for neurodegenerative disorders expressing abnormally high brain levels of p25 and hyperactive cdk5.

Broad Spectrum Anti-Quorum Sensing Activity of Tannin-Rich Crude Extracts of Indian Medicinal Plants.

Quorum sensing (QS) mechanisms have been demonstrated to have significance in expression of pathogenicity in infectious bacteria. In Gram negative bacteria the autoinducer molecules that mediate QS are acyl homoserine lactones (AHL) and in Gram positive bacteria they are peptides called autoinducing peptides (AIP). A screening of tannin-rich medicinal plants was attempted to identify extracts that could interrupt the QS mechanisms in both Gram positive and Gram negative bacteria over a wide range of concentrations and therefore potentially be potent agents that could act as broad spectrum QS inhibitors. Six out of the twelve Indian medicinal plant extracts that were analyzed exhibited anti-QS activity in Chromobacterium violaceum 12472 and in S. aureus strain with agr:blaZ fusion over a broad range of subinhibitory concentrations, indicating that the extracts contain high concentration of molecules that can interfere with the QS mechanisms mediated by AHL as well as AIP.

The interaction of Munc 18 (p67) with the p10 domain of p35 protects in vivo Cdk5/p35 activity from inhibition by TFP5, a peptide derived from p35.

In a series of studies, we have identified TFP5, a truncated fragment of p35, the Cdk5 kinase regulatory protein, which inhibits Cdk5/p35 and the hyperactive Cdk5/p25 activities in test tube experiments. In cortical neurons, however, and in vivo in Alzheimer's disease (AD) model mice, the peptide specifically inhibits the Cdk5/p25 complex and not the endogenous Cdk5/p35. To account for the selective inhibition of Cdk5/p25 activity, we propose that the "p10" N-terminal domain of p35, absent in p25, spares Cdk5/p35 because p10 binds to macromolecules (e.g., tubulin and actin) as a membrane-bound multimeric complex that favors p35 binding to Cdk5 and catalysis. To test this hypothesis, we focused on Munc 18, a key synapse-associated neuronal protein, one of many proteins copurifying with Cdk5/p35 in membrane-bound multimeric complexes. Here we show that, in vitro, the addition of p67 protects Cdk5/p35 and has no effect on Cdk5/p25 activity in the presence of TFP5. In cortical neurons transfected with p67siRNA, we also show that TFP5 inhibits Cdk5/p35 activity, whereas in the presence of p67 the activity is protected. It does so without affecting any other kinases of the Cdk family of cyclin kinases. This difference may be of significant therapeutic value because the accumulation of the deregulated, hyperactive Cdk5/p25 complex in human brains has been implicated in pathology of AD and other neurodegenerative disorders.

Profiling of p5, a 24 Amino Acid Inhibitory Peptide Derived from the CDK5 Activator, p35 CDKR1 Against 70 Protein Kinases.

Cyclin-dependent kinase 5 (CDK5) is a multifunctional serine/threonine kinase that regulates a large number of neuronal processes essential for nervous system development and function with its activator p35 CDK5R1. Upon neuronal insults, p35 is proteolyzed and cleaved to p25 producing deregulation and hyperactivation of CDK5 (CDK5/p25), implicated in tau hyperphosphorylation, a pathology in some neurodegenerative diseases. A truncated, 24 amino acid peptide, p5, derived from p35 inhibits the deregulated CDK5 phosphotransferase activity and ameliorates Alzheimer's disease (AD) phenotypes in AD model mice. In the present study, we have screened a diverse panel of 70 human protein kinases for their sensitivities to p5, and a subset of these to p35. At least 16 of the tested protein kinases exhibited IC50 values that were 250 µM or less, with CAMK4, ZAP70, SGK1, and PIM1 showing greater sensitivity to inhibition by p5 than CDK5/p35 and CDK5/p25. In contrast, the p5 peptide modestly activated LKB1 and GSK3ß. A sub set of kinases screened against p35 showed that activity of CAMK4 in the absence of calcium and calmodulin was also markedly inhibited by p35. The Cyclin Y-dependent kinases PFTK1 (CDK14) and PCTK1 (CDK16) were activated by p35 at least 10-fold in the absence of Cyclin Y and by approximately 50% in its presence. These findings provide additional insights into the mechanisms of action for p5 and p35 in the regulation of protein phosphorylation in the nervous system.

Knockdown of Expression of Cdk5 or p35 (a Cdk5 Activator) Results in Podocyte Apoptosis.

Podocytes are terminally differentiated glomerular epithelial cells. Podocyte loss has been found in many renal diseases. Cdk5 is a cyclin-dependent protein kinase which is predominantly regulated by p35. To study the role of Cdk5/p35 in podocyte survival, we first applied western blotting (WB) analysis to confirm the time-course expression of Cdk5 and p35 during kidney development and in cultured immortalized mouse podocytes. We also demonstrated that p35 plays an important role in promoting podocyte differentiation by overexpression of p35 in podocytes. To deregulate the expression of Cdk5 or p35 in mouse podocytes, we used RNAi and analyzed cell function and apoptosis assaying for podocyte specific marker Wilms Tumor 1 (WT1) and cleaved caspase 3, respectively. We also counted viable cells using cell counting kit-8. We found that depletion of Cdk5 causes decreased expression of WT1 and apoptosis. It is noteworthy, however, that downregulation of p35 reduced Cdk5 activity, but had no effect on cleaved caspase 3 expression. It did, however, reduce expression of WT1, a transcription factor, and produced podocyte dysmorphism. On the other hand increased apoptosis could be detected in p35-deregulated podocytes using the TUNEL analysis and immunofluorescent staining with cleaved caspase3 antibody. Viability of podocytes was decreased in both Cdk5 and p35 knockdown cells. Knocking down Cdk5 or p35 gene by RNAi does not affect the cycline I expression, another Cdk5 activator in podocyes. We conclude that Cdk5 and p35 play a crucial role in maintaining podocyte differentiation and survival, and suggest these proteins as targets for therapeutic intervention in podocyte-damaged kidney diseases.

Sustained release of a purified tannin component of Terminalia chebula from a titanium implant surface prevents biofilm formation by Staphylococcus aureus.

Although biofilms are formed on a variety of surfaces, of utmost significance are those formed on prosthetic devices used as implants. Such biofilms can lead to severe device-related infections that are difficult to treat. In a search for new antibiofilm agents that can be used as "active" implant coatings, purified fraction from a tannin-rich extract of Terminalia chebula was isolated and tested for its antibiofilm properties on a titanium implant material. The fraction, named as Fraction 7, was found to significantly reduce biofilm formation by hospital isolates of Staphylococcus aureus, at sub-inhibitory concentrations that were 64 times lower than the minimum inhibitory concentration (MIC). Simulated local delivery systems of the Fraction 7 set upon the surface of titanium alloy released the fraction in a controlled manner from a biodegradable carrier (PDLLA) and were found to significantly reduce biofilm formation by a methicillin-resistant hospital isolate of S. aureus in a load concentration dependent manner without preventing growth. This study therefore identifies a novel fraction from tannin-rich extract of T. chebula that has potential to be used as an antibiofilm coat on implant surfaces.

Antimicrobial Susceptibility Pattern of Bacterial Isolates in Burn Patients in a Tertiary Care Center: A Prospective Study

Background: Burn injuries and deaths pose a major public health concern globally, especially in developing and underdevelopedcountries. As burn patients have lost their primary barrier and exposed to microorganism invasion continually and chronically,Staphylococcus aureus becomes one of the greatest causes of nosocomial infection in burn patients though it is a normalskin flora. The cases of antibiotic resistance have increased, and resistant species such as methicillin-resistant S. aureus(MRSA) provide additional challenges in the form of virulence factors. Multimodal infection control concept is required to limitthe spread of infection with multidrug-resistant organism including MRSA in a burn unit. The common pathogens isolated fromburn wounds are S. aureus, Pseudomonas aeruginosa, Streptococcus pyogenes, Acinetobacter baumannii, and various coliformbacilli. Hence, antimicrobial susceptibility pattern of bacterial isolates in burn patients plays a key role in the management ofthese patients.Materials and Methods: This prospective observational study involved the collection of wound swabs from burn patientsfrom June 2018 to May 2019. All patients with burn wounds irrespective of age and sex, admitted through surgery outpatientdepartment or casualty, during the period of study were included in the study.Results: Maximum prevalence was found for P. aeruginosa, i.e., 37.5% followed by S. aureus, for which the prevalencewas found to be 18.75%. The organism least commonly cultured was Acinetobacter; the prevalence of MRSA was found tobe 57.14% and the prevalence of methicillin resistance was found to be 42.8% in patients with Staphylococcus epidermidis.Overall, the prevalence of methicillin resistance was 51.72%. The drugs most effective against P. aeruginosa, the most commonisolate, were meropenem (97.62%) and piperacillin/tazobactam (90.48%) followed by gentamicin (64.29%). Meropenem andpiperacillin/tazobactam showed 100% efficacy against the other Gram-negative bacilli isolated as well. MRSA isolates showed100% sensitivity to vancomycin and linezolid closely followed by piperacillin-tazobactam combination. Klebsiella pneumoniaeshowed 100% sensitivity to meropenem and piperacillin/tazobactam.Conclusions: The overall isolation rate was 75%. Only solitary isolates were studied. Overall, Gram-negative organisms(66.66%) were more common than Gram-positive organisms (33.33%). P. aeruginosa (37.5%) was the most commonisolate followed by S. aureus (18.75%). The prevalence of MRSA was 57.14%, but all the MRSA isolates showed 100%sensitivity to vancomycin and linezolid. On antibiotic sensitivity testing, piperacillin/tazobactam (95.24%) was found to bethe most effective drug against all the organisms isolated. Meropenem (99.40%) was the most effective drug against theGram-negative organisms. Vancomycin (100%) and linezolid (100%) were the most effective drugs for the Gram-positiveorganisms.

A Clinical Study of Effect of Centchroman on Fibroadenoma

Background: Centchroman has been studied earlier and literature mentions it to be a novel non-steroidal, selective estrogenreceptor modulator, antiestrogen, and mild anti-inflammatory drug with a significant decrease in size of fibroadenoma. Ourstudy aims to check the reduction in the size of fibroadenoma in response to centchroman.Materials and Methods: A prospective observational study was carried out in patients of fibroadenoma attending surgeryoutpatient department and wards of Sanjay Gandhi Memorial Hospital associated with Shyam Shah Medical College, Rewa(Madhya Pradesh) from August 2015 to July 2016. Patients were included in the study after obtaining an informed consent.Patients were followed up for 12 weeks.Results: In our study, the maximum number of fibroadenomas was found in the left upper outer quadrant; of 102 patients,total lesions (fibroadenoma) were 130. Of 102 patients studied for the effect of centchroman on fibroadenoma, there was aresponse in 36 patients, which accounted for 35.29%. In 66 (64.8%) patients, there was no response after the treatment withcentchroman and they were subjected to excision and biopsy. The mean difference in volumes of fibroadenoma was statisticallyinsignificant. However, there was a reduction in size of mean volume of fibroadenoma which was 4.085 at the presentation andwhich was 3.24 at the end of the 12th week.Conclusion: Fibroadenoma was common in the age group of 21–30 years. The left breast was more involved in fibroadenomas.The most common site for fibroadenoma was the left upper outer quadrant. The effect of centchroman on decrease in volume offibroadenoma was seen in 35.29% of patients. Reduction in volume of fibroadenoma was statistically insignificant. More than 50%reduction in volume of fibroadenoma was seen only in 3 patients (2.94%). Surgical excision and biopsy were the preferred modalityof treatment for fibroadenomas in patients where the drug centchroman showed no response in regression in volume (64.8%).

Methicillin-resistant Staphylococcus aureus-prevalence and Antimicrobial Sensitivity Pattern among Burn Patients in a Tertiary Care Hospital in Vindhya Region

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a well-recognized public health problem throughout theworld. The evolution of new genetically distinct community-acquired and livestock-acquired MRSA and extended resistance toother non-β-lactams including vancomycin has only amplified the crisis. This paper presents data on the prevalence of MRSAand resistance pattern to other antibiotics on the selected specimen from burn patients.Materials and Methods: This is a prospective study conducted in the burn unit of Shyam Shah Medical College and SanjayGandhi Memorial Hospital, Rewa (M.P.), from June 2018 to May 2019, where all patients with flame and scald burns wereincluded in the study who had up to a second degree or partial-thickness burns.Results: A total of 558 patients were admitted in the burn unit throu`ghout the year, the age ranged from 2 months to 85 years.About 56.10% were females and 43.90% were males. Pseudomonas aeruginosa (37.5%) was the most common isolatefollowed by S. aureus (18.75%). The prevalence of MRSA was 57.14% but all the MRSA isolates showed 100% sensitivity tovancomycin and linezolid closely followed by piperacillin and tazobactam combination. The prevalence of methicillin resistanceoverall among S. aureus and Staphylococcus epidermidis was found to be 51.72%.Conclusion: MRSA is prevalent among the burn wounds but is 100% sensitive to vancomycin and linezolid. To ensure earlyand appropriate therapy, routine microbiological surveillance and a regular update of their antimicrobial susceptibility patterncould help in the prevention of development of multidrug resistance.

A Prospective Study of the Demographic Pattern and Site of Perforation of Non-traumatic Hollow Viscus Perforation Peritonitis in Vindhya Region

Background: Perforation peritonitis is a commonly encountered surgical emergency and it is defined as inflammation of theserosal membrane that lines the abdominal cavity and the visceral organs. The aim of this study is to analyze the demographicpattern and site of perforation of non-traumatic hollow viscus perforation peritonitis in Vindhya region.Materials and Methods: A total of 209 cases were studied with hollow viscus perforation peritonitis admitted in the surgicalwards in Sanjay Gandhi Memorial Hospital associated with Shyam Shah Medical College, Rewa (MP), India, in the period fromJune 1, 2018, to May 31, 2019. All necessary investigations were carried out. X-ray, Ultrasonography abdomen, and bloodinvestigations were done. The patient underwent emergency exploratory laparotomy and a careful record of pre-operative andpost-operative findings was made and was carefully filled in the pro forma. All the patients were advised to attend the surgicaloutpatient department for follow-up.Results: Of 10,887 patients admitted to Sanjay Gandhi Memorial Hospital associated with Shyam Shah Medical College, Rewa(MP), India, from June 1, 2018, to May 31, 2019, in which non-traumatic hollow viscus perforation peritonitis was diagnosedin 209 patients (1.9%), among which most of the patients were male (177) and rest were female (32). Most of the patientsbelonged to the low-socio-economic status of 21–40 years of the age group. From this study, the duodenum was found to bethe most common site of perforation, followed by stomach.Conclusion: Patients were admitted in the Department of Surgery, Shyam Shah Medical College and Sanjay Gandhi HospitalRewa, the Vindhya region in the Madhya Pradesh, patients diagnosed as a case of non-traumatic hollow viscus perforationperitonitis were included in the study. The majority of the patients of the perforation peritonitis belonged to 21–40 years ofage group. 41–60 years of age group was the second most common age group of patients who presented with perforationperitonitis with a male-to-female ratio of 5.5:1. The most common site of perforation was duodenum followed by gastric andappendicular and the least common site of perforation was colon.

Analysis of the Inhibitory Elements in the p5 Peptide Fragment of the CDK5 Activator, p35, CDKR1 Protein.

Besides the hallmark pathology of amyloid plaques and neurofibrillary tangles, it is well documented that cyclin-dependent kinase 5 (CDK5), a critical neuronal protein kinase in nervous system development, function, and survival, when deregulated and hyperactivated induces Alzheimer's disease (AD) and amyotrophic lateral sclerosis and Parkinson's disease-like phenotypes in mice. In a recent study, we demonstrated that p5, a small, truncated fragment of 24 amino acid residues derived from the CDK5 activator protein 35 (NCK5A, p35), selectively inhibited deregulated CDK5 hyperactivity and ameliorated AD phenotypes in model mice. In this study, we identified the most inhibitory elements in the p5 peptide fragment. Each amino acid residue in p5 was systematically replaced with its homologous residues that may still be able to functionally substitute. The effects of these p5 peptide analogs were studied on the phosphotransferase activities of CDK5/p35, CDK5/p25, ERK1, and GSK3ß. The mimetic p5 peptide (A/V substitution at the C-terminus of the peptide) in the sequence, KNAFYERALSIINLMTSKMVQINV (p5-MT) was the most effective inhibitor of CDK5 kinase activity of 79 tested mimetic peptides including the original p5 peptide, KEAFWDRCLSVINLMSSKMLQINA (p5-WT). Replacement of the residues in C-terminus end of the peptide affected CDK5 phosphotransferase activity most significantly. These peptides were strong inhibitors of CDK5, but not the related proline-directed kinases, ERK1 and GSK3ß.

Peptide TFP5/TP5 derived from Cdk5 activator P35 provides neuroprotection in the MPTP model of Parkinson's disease.

Parkinson's disease (PD) is a chronic neurodegenerative disorder characterized by the loss of dopamine neurons in the substantia nigra, decreased striatal dopamine levels, and consequent extrapyramidal motor dysfunction. Recent evidence indicates that cyclin-dependent kinase 5 (Cdk5) is inappropriately activated in several neurodegenerative conditions, including PD. To date, strategies to specifically inhibit Cdk5 hyperactivity have not been successful without affecting normal Cdk5 activity. Previously we reported that TFP5 peptide has neuroprotective effects in animal models of Alzheimer's disease. Here we show that TFP5/TP5 selective inhibition of Cdk5/p25 hyperactivation in vivo and in vitro rescues nigrostriatal dopaminergic neurodegeneration induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP/MPP+) in a mouse model of PD. TP5 peptide treatment also blocked dopamine depletion in the striatum and improved gait dysfunction after MPTP administration. The neuroprotective effect of TFP5/TP5 peptide is also associated with marked reduction in neuroinflammation and apoptosis. Here we show selective inhibition of Cdk5/p25 -hyperactivation by TFP5/TP5 peptide, which identifies the kinase as a potential therapeutic target to reduce neurodegeneration in Parkinson's disease.