It's all relative: analyzing microbiome compositions, its significance, pathogenesis and microbiota derived biofilms: Challenges and opportunities for disease intervention.

Concept of microorganisms has largely been perceived from their pathogenic view point. Nevertheless, it is being gradually revisited in terms of its significance to human health and now appears to be the most dominant force that shapes the immune system of the human body and also determines an individual's predisposition to diseases. Human inhabits bacterial diversity (which is predominant among all microbial communities in human body) occupying 0.3% of body mass, known as microbiota. On birth, a part of microbiota that child obtains is essentially a mother's legacy. So, the review was initiated with this critical topic of microbiotal inheritance. Since, each body site has distinct physiological specifications; therefore, they contain discrete microbiome composition that has been separately discussed along with dysbiosis-induced pathologies originating in different body organs. Factors affecting microbiome composition and may cause dysbiosis like antibiotics, delivery, feeding method etc. as well as the strategies that immune system adopts to prevent dysbiosis have been highlighted. We also tried to bring into attention the topic of dysbiosis induced biofilms, that enables cohort to survive stresses, evolve, disseminate and infection resurgence that is still in dormancy. Eventually, we put spotlight on microbiome significance in medical therapeutics. We didn't merely confine article to gut microbiota, that is being studied more extensively. Numerous community forms at diverse body sites are inter-related, and being exposed to awfully variable perturbations appear to be challenging to evaluate perturbation risks holistically. All aspects have been elaborately discussed to achieve a global depiction of human microbiota in order to meet urgent necessity for protocol standardisation. Demonstrates that environmental challenges (antibiotic use, alterations in diet, stress, smoking etc.) might cause dysbiosis i.e. transition of healthy microbiome composition to the one in which pathogenic microorganisms become more abundant, and eventually results in an infected state.

Immediate and Long-Term Effectiveness of Proprioceptive Neuromuscular Facilitation and Static Stretching on Joint Range of Motion, Flexibility, and Electromyographic Activity of Knee Muscles in Older Adults.

Introduction and Objective: Previously, various stretching techniques were compared to study their effects on the different physiological parameters of hamstring muscles in the elderly population. There is no research that compares the immediate and long-term effects of proprioceptive neuromuscular facilitation-contract-relax (PNF-CR) and static stretching (SS) techniques on knee range of motion (ROM), hamstring flexibility, and knee flexor muscle EMG activity in the elderly. This study intends to compare the same. Methods: A total of 30 males aged 55-75 years were randomly assigned into the PNF-CR group (n = 10), SS group (n = 10), and control group (n = 10). The PNF-CR group received four trials of the contract-relax technique, the SS group received passive stretching of an 80 s duration by the therapist, and the control group received no intervention. A total of 12 sessions were given during the four-week period. Knee range of motion, electromyographic activity of the biceps femoris, and the sit-and-reach test were taken for the dominant side thrice: pre-intervention, immediately after stretching, and after the training period. Results: A statistically significant difference was observed in the maximum voluntary isometric contraction (MVIC) of biceps femoris between the PNF and the control groups (p = 0.01) after four weeks of intervention. The knee ROM and hamstring flexibility for the PNF group showed significant improvement immediately post-test (p = 0.01) and after four weeks of training (p = 0.07 and p = 0.001). SS showed significant results for both ROM and flexibility after four weeks of intervention (p = 0.001), and significant immediate post-test improvements were seen for ROM only (p = 0.007). Conclusions: PNF stretching has an immediate, as well as long-term, effect on knee ROM and hamstring flexibility, whereas it has only a long-term effect on muscle electromyographic activity. SS has an immediate, as well as long-term, effect on knee ROM and only a long-term effect on hamstring flexibility, without any immediate or long-term effects on muscle electromyographic activity.

Establishing Responsiveness and Minimal Clinically Important Difference of Quebec Back Pain Disability Scale (Hindi Version) in Chronic Low Back Pain Patients Undergoing Multimodal Physical Therapy.

Increasing emphasis is placed on physical functional measures to examine treatments for chronic low back pain (CLBP). The Quebec Back Pain Disability Scale (Hindi version) (QBPDS-H) has never been evaluated for responsiveness. The objectives of this study were to (1) examine the internal and external responsiveness of the Quebec Back Pain Disability Scale (Hindi version) (QBPDS-H) and (2) find out the minimal clinically important difference (MCID) and minimal detectable change (MDC) in the functional ability of patients with chronic low back pain (CLBP) undergoing multimodal physical therapy treatment. In this prospective cohort study, QBPDS-H responses were recorded at the baseline and after eight weeks from 156 CLBP patients undergoing multimodal physiotherapy treatment. To differentiate between the clinically unimproved (n = 65, age: 44.16 ± 11.8 years) and clinically improved (n = 91, age: 43.28 ± 10.7 years) scores of patients from the initial assessment to the last follow-up, the Hindi version of the Patient's Global Impression of Change (H-PGIC) scale was utilized. Internal responsiveness was large (E.S. (pooled S.D.) (n = 91): 0.98 (95% CI = 1.14-0.85) and Standardized Response Mean (S.R.M.) (n = 91): 2.57 (95% CI = 3.05-2.17)). In addition, the correlation coefficient and receiver operative characteristics (R.O.C.) curve were used to assess the QBPDS-H external responsiveness. MCID and MDC were detected by the R.O.C. curve and standard error of measurements (S.E.M.), respectively. The H-PGIC scale showed moderate responsiveness (ρ = 0.514 and area under the curve (A.U.C.) = 0.658; 95% CI, 0.596-0.874), while the MDC achieved 13.68 points, and the MCID was found have 6 points (A.U.C. = 0.82; 95% CI: 0.74-0.88, sensitivity = 90%, specificity = 61%). This study shows that QBPDS-H has moderate levels of responsiveness in CLBP patients receiving multimodal physical therapy treatment, so it can be used to measure the changes in disability scores. MCID and MDC changes were also reported with QBPDS-H.

mltG gene deletion mitigated virulence potential of Streptococcus mutans: An in-vitro, ex-situ and in-vivo study.

Bacterial cells are surrounded by a peptidoglycan (PG) cell wall, which is essential for cell integrity and intrinsic biogenesis pathways; hence, the cell wall is a potential target for several antibiotics. Among several lytic transglycosylases (LTs), the mltG gene plays a crucial role in the synthesis of peripheral PG. It localises the re-modelled PGs for septum formation and cleavage across the bacterial cell wall during daughter cells separation. However, the role of mltG gene in bacterial virulence, particularly in Gram-positive bacteria during dentine biofilm and caries development, has remained unexplored. Hence, we exploited Gram-positive Streptococcus mutans cells for the very first time to construct a mltG knock-out bacterial strain, e.g., ΔmltG S. mutans. Systematic comparative investigations revealed that doubling time (Td), survival, enzymatic efficiencies, pH tolerance, bio-synthesise of lipid, proteins and DNA, biofilm formation and dentine lesions were significantly (p < 0.001) compromised in case of ΔmltG S. mutans than wild type strain. The qRT-PCR based gene expression profiling revealed that transcriptional expression of critically important genes involved in biofilm, metabolism, and stress response were dysregulated in the mutant. Besides, an incredible reduction in dentine caries development was found in the molar teeth of Wistar rats and also in human extracted teeth. Concisely, these trends obtained evidently advocated the fact that the deletion of mltG gene can be a potential target to impair the S. mutans virulence through severe growth retardation, thereby reducing the virulence potential of S. mutans.

Orally fed EGCG coronate food released TiO2 and enhanced penetrability into body organs via gut.

Owing to unique nano-scale properties, TiO2-NPs (T-NPs) are employed as food-quality enhancers in >900 processed food products. Whereas, epigallocatechin-3-gallate (EGCG), a green tea polyphenol is consumed in traditional brewed tea, globally. Taken together, we aimed to investigate whether human gastric-acid digested T-NPs and complex tea catechins yield ionic species (Ti4+, Ti3+ etc.) and active EGCG forms to meet favourable conditions for in vivo bio-genesis of EGCG-coronated TiO2-NPs (ET-NPs) in human gut. Secondly, compared to bare-surface micro and nano-scale TiO2, i.e., T-MPs and T-NPs, respectively, how EGCG coronation on ET-NPs in the gut facilitates the modulation of intrinsic propensity of internalization of TiO2 species into bacteria, body-organs, and gut-microbiota (GM), and immune system. ET-NPs were synthesized in non-toxic aqueous solution at varied pH (3-10) and characterised by state-of-the-arts for crystallinity, surface-charge, EGCG-encapsulation, stability, size, composition and morphology. Besides, flow-cytometry (FCM), TEM, EDS, histopathology, RT-PCR, 16S-rRNA metagenomics and ELISA were also performed to assess the size and surface dependent activities of ET-NPs, T-NPs and T-MPs vis-a-vis planktonic bacteria, biofilm, GM bacterial communities and animal's organs. Electron-microscopic, NMR, FTIR, DLS, XRD and EDS confirmed the EGCG coronation, dispersity, size-stability of ET-NPs, crystallinity and elemental composition of ET-NPs-8 and T-NPs. Besides, FCM, RT-PCR, 16S-rRNA metagenomics, histopathology, SEM and EDS analyses exhibited that EGCG coronation in ET-NPs-8 enhanced the penetration into body organs (i.e., liver and kidney etc.) and metabolically active bacterial communities of GM.

Protein expression profiling, in silico classification and pathway analysis of cariogenic bacteria Streptococcus mutans under bacitracin stress conditions.

Introduction. Streptococcus mutans is a cariogenic bacterium that causes dental caries as well as being implicated in other dental pathologies and infective endocarditis. Bacitracin is a bactericidal antibiotic that induces cell wall stress in Gram-positive bacteria.Gap Statement. S. mutans is among the most characterized Gram-positive bacteria. However, the transcriptome and proteome of S. mutans have received less attention, and they are actually key in understanding the pathogenesis of any bacteria. In this study, we extracted the whole proteome of S. mutans grown under bacitracin stress. Such a proteome is anticipated to offer deep insights related to physiological dynamic fluctuations and, consequently, it may provide 'proteomic signatures' to be identified as potential targets.Aim. The aim of the study is to explore the general stress response that S. mutans exhibits at the proteome level when cell wall stress is imposed on it.Methodology. A sub-MIC concentration of bacitracin was added to the growth media of S. mutans followed by whole-cell protein extraction. The proteome was then subjected to high-throughput proteomics analysis, i.e. liquid chromatography tandem mass spectrometry (LC-MS/MS). Differentially expressed proteins obtained through LC-MS/MS underwent analyses such as gene ontology, KEGG (Kyoto Encyclopaedia of Genes and Genomes) and DAVID (Database for Annotation, Visualization and Integrated Discovery) analysis, and STRING for functional annotation, pathway enrichment and protein-protein interaction (PPI) networks, respectively. These proteins were also categorized into functional classes using the PANTHER (Protein Annotation Through Evolutionary Relationship) classification system.Result. LC-MS/MS produced data from 321 identified proteins. From these, 41 and 30 were found to be significantly over- (≥2 fold change) and underexpressed (≤0.4 fold change), respectively. In the upregulated proteins we mostly observed sortases and proteins involved in the EPS biosynthesis pathway, whereas among the downregulated proteins the majority related to glycolysis.Conclusion. The sortase family of proteins appear to be potential targets because they regulate various virulence factors and therefore can be targeted to inhibit multiple virulence pathways simultaneously. This study offers an understanding of proteomic fluctuations in response to cell wall stress and can thus help in identifying key players mediating virulence.

Effects of scapular stabilization and upper limb proprioception as an adjunct to cervical stabilization in chronic neck pain patients: A randomized controlled trial.

BACKGROUND: Chronic neck pain is a prevalent health condition and a leading cause of disability worldwide. Prompt therapeutic measures are required to overcome this condition. OBJECTIVES: To evaluate the efficacy of incorporation of scapular stabilization and upper limb proprioceptive exercises to cervical stabilization exercises in patients with chronic neck pain (CNP). DESIGN: A single-blinded randomized controlled design. METHODS: A sample of convenience was deployed to recruit twenty-eight patients having CNP (18-45 years) and was randomized into two groups: group A (cervical stabilization exercises group) and group B (scapular stabilization and upper limb proprioceptive exercises group + cervical stabilization exercises). Pain intensity, disability, sleep quality, quality of life, scapular muscles strength and proprioception were assessed at 4 weeks follow up to determine the efficacy of the intervention. RESULTS: A mixed model ANOVA was used. A statistically significant (p < 0.05) group by time interaction for pain intensity (p = 0.000), scapular muscles strength of all muscles (p = 0.000) was observed. Significant group interaction for absolute error (p = 0.00), for pain (p = 0.001), disability (p = 0.04) and scapular muscle's strength (p = 0.000) was also demonstrated. CONCLUSION: The results indicated that scapular stabilization and upper limb proprioceptive exercises when combined with cervical stabilization exercises are more beneficial in alleviating pain and disability and improving scapular muscle strength and proprioception in patients with CNP.

Proteomic Characterization and Target Identification Against Streptococcus mutans Under Bacitracin Stress Conditions Using LC-MS and Subtractive Proteomics.

The aim of the present study, is to identify potential targets against the highly pathogenic bacteria Streptococcus mutans that causes dental caries as well as the deadly infection of endocarditis. The powerful and highly sensitive technique of liquid chromatography-mass spectrometry (LC-MS/MS) identified 321 proteins of S. mutans when grown under stressful conditions induced by the antibiotic bacitracin. These 321 proteins were subjected to the insilico method of subtractive proteomics to screen out potential targets by utilizing different analyses like CD-HIT, non-homologous sequence screening, KEGG pathway, essentiality screening, gut-flora non-homology, and codon usage analysis. A database of essential proteins was employed to find sequence homology of non-paralogous proteins to determine proteins which are essential for bacterial survival. Cellular localization analysis of the selected proteins was done to localize them inside the cell along with physico-chemical characterization and druggability analysis. Using computational tools, 22 proteins out of 321, that are functionally distinguishable from their human counterparts and passed the criterion of a potential therapeutic candidate were identified. The selected proteins comprise central energy metabolic proteins, virulence factors, proteins of the sortase family, and essentiality factors. The presented analyses identified proteins of the sortase family, which appear as key therapeutic targets against caries infection. These proteins regulate a number of virulence factors, thus can be simultaneously inhibited to obstruct multiple virulence pathways.

Exploring antibiofilm potential of bacitracin against streptococcus mutans.

Bacitracin has well familiar effects on growth and colonization of bacteria but its antibiofilm action on majority of bacteria is still not studied. Bacitracin is a bactericidal antibiotic that primarily acts on Gram positive bacteria by obstructing the process of cell wall synthesis. In this study, we have investigated antibiofilm potential and the mechanism of bacitracin against a cariogenic bacteria 'Streptococcus mutans' which has not been reported so far. Bacitracin has been found to affect propensity of S. mutans to form biofilm. On treatment with sub-MIC concentration of bacitracin resulted in significant reduction in bifilm formation as evaluated by crystal violet and congo red assays. The architecture of S. mutans biofilm was observed by scanning electron microscopy which revealed astonishing phenotype of biofilm. Deficient biofilm was found to be composed of abnormally elongated cells. Transmission electron microscopy showed multiple septa formation in each cell of biofilm thereby indicating, cell division defect as the most probable cause of cell elongation. To elucidate the effect of bacitracin on molecular level, expression profiling of genes critically important for cell division and biofilm formation was performed, which were found many folds downregulated. Bacitracin at very low concentration has been found to have potent antibiofilm activity, therefore is a potential antibiofilm agent to treat oral biofilms. It is being anticipated, this study will offer novel information to identify potential targets and effectively creates true innovation to understand the biofilm's basic biology. Besides, discovering new uses for currently marketed drugs makes commercial as well as research sense.

Efficacy of photodynamic therapy against Streptococcus mutans biofilm: Role of singlet oxygen.

In photodynamic therapy (PDT), killing is entirely based on the ROS generation and among different types of ROS generated during PDT, singlet oxygen is considered as the most potential as illustrated in many studies and therefore it is predominantly responsible for photodamage and cytotoxic reactions. The aim of this study was to check whether singlet oxygen (Type II photochemistry) is more potential than free radicals (Type I photochemistry) against Streptococcus mutans biofilm. We have taken two phenothiazinium dyes i.e. toluidine blue O (TBO) and new methylene blue (NMB). TBO was found to have better antibacterial as well as antibiofilm effect than NMB. Antibacterial effect was evaluated by colony forming unit while antibiofilm action by crystal violet and congo red binding assays. We have also evaluated the disruption of preformed biofilm by biofilm reduction assay, confocal laser electron and scanning electron microscopy. More singlet oxygen production was detected in case of TBO than NMB while more Free radical (HO) was produced by NMB than TBO. TBO showed better antibacterial as well as antibiofilm effect than NMB so; we conclude that potency of a photosensitizer is correlated with the capability to produce singlet oxygen.

Transcultural adaptation and validation of Hindi version of Quebec Back Pain Disability Scale.

PURPOSE: To transculturally adapt the Quebec Back Pain Disability Scale for Hindi-speaking population and examine its psychometric properties in patients with low back pain. MATERIALS AND METHODS: The Quebec Back Pain Disability Scale was translated and cross-culturally adapted into Hindi following international guidelines. Hindi version of the scale was completed by 120 patients with low back pain and 60 healthy controls. Patients with low back pain were also administered the Hindi-Roland Morris Disability Questionnaire and Visual Analog Scale. Psychometric evaluation included test-retest reliability, convergent and discriminative validity. Exploratory factor analysis was carried out to determine the factor structure. RESULTS: The factorial analysis revealed a four-factor solution (bending/carrying, ambulation/reach, prolonged postures and rest). Convergent validity was confirmed by high correlation of Hindi Quebec Back Pain Disability Scale to the Hindi version of Roland Morris Disability Questionnaire (r = 0.77 and p < 0.001) as well as Visual Analog Scale (r = 0.682 and p < 0.001) scores. Discriminative validity was established by significantly different scores for patients with low back pain and the healthy controls (35.36 ± 18.6 vs. 9.13 ± 6.08 and p < 0.001). The translated version of the scale showed remarkable internal consistency (Cronbach α = 0.98) and the intraclass correlation coefficient of test-retest reliability was excellent (ICC2,1=0.96). MDC95 and SEM scores obtained were 10.28 and 3.71, respectively. CONCLUSION: The Hindi version of Quebec Back Pain Disability Scale has good test-retest reliability, discriminative and convergent validity and is appropriate for clinical and research use in Hindi-speaking low back pain patients. Implications for rehabilitation Linguistically and culturally adapted questionnaires help researchers make adequate inferences about instruments measuring health and quality of life. The translated version would serve as a valid research tool allowing comparability of data across cultures thus, providing opportunities for large multicenter, multicountry trials. A Hindi Quebec Back Pain Disability Scale version will help to improve the quality and efficacy of assessment of low back pain by developing in patients, a better understanding of the items which can be easily correlated with the activities of daily living.

Nano-therapeutics: A revolution in infection control in post antibiotic era.

With the arrival of antibiotics 70 years ago, meant a paradigm shift in overcoming infectious diseases. For decades, drugs have been used to treat different infections. However, with time bacteria have become resistant to multiple antibiotics, making some diseases difficult to fight. Nanoparticles (NPs) as antibacterial agents appear to have potential to overcome such problems and to revolutionize the diagnosis and treatment of bacterial infections. Therefore, there is significant interest in the use of NPs to treat variety of infections, particularly caused by multidrug-resistant (MDR) strains. This review begins with illustration of types of NPs followed by the literature of current research addressing mechanisms of NPs antibacterial activity, steps involved in NP mediated drug delivery as well as areas where NPs use has potential to improve the treatment, like NP enabled vaccination. Besides, recently emerged innovative NP platforms have been highlighted and their progress made in each area has been reviewed.

A comparison of antibacterial and antibiofilm efficacy of phenothiazinium dyes between Gram positive and Gram negative bacterial biofilm.

BACKGROUND: Antimicrobial photodynamic therapy (APDT) is a process that generates reactive oxygen species (ROS) in presence of photosensitizer, visible light and oxygen which destroys the bacterial cells. We investigated the photoinactivation efficiency of phenothiazinium dyes and the effect of ROS generation on Gram positive and Gram negative bacterial cell as well as on biofilm. MATERIAL AND METHODS: Enterococcus faecalis and Klebsiella pneumonia were incubated with all the three phenothiazinium dyes and exposed to 630nm of light. After PDT, colony forming unit (CFU) were performed to estimate the cell survival fraction. Intracellular reactive oxygen species (ROS) was detected by DCFH-DA. Crystal violet (CV) assay and extracellular polysaccharides (EPS) reduction assay were performed to analyze antibiofilm effect. Confocal laser electron microscope (CLSM) scanning electron microscope (SEM) was performed to assess the disruption of biofilm. RESULTS: 8log10 reduction in bacterial count was observed in Enterococcus faecalis while 3log10 in Klebsiella pneumoniae. CV and EPS reduction assay revealed that photodynamic inhibition was more pronounced in Enterococcus faecalis. In addition to this CLSM and SEM study showed an increase in cell permeability of propidium iodide and leakage of cellular constituents in treated preformed biofilm which reflects the antibiofilm action of photodynamic therapy. CONCLUSION: We conclude that Gram-positive bacteria (Enterococcus faecalis) are more susceptible to APDT due to increased level of ROS generation inside the cell, higher photosensitizer binding efficiency and DNA degradation. Phenothiazinium dyes are proved to be highly efficient against both planktonic and biofilm state of cells.

Efficacy of Complementary Therapies in the Quality of Life of Breast Cancer Survivors.

Breast cancer (BC) is the most common cancer diagnosed in women and the second most common cancer overall, ranking as the fifth cause of death from cancer. The chronicity of the disease produces long-term physiological and psychological manifestations, which adversely affect the quality of life of the individual. The primary treatment while managing cancer presents with various debilitating side effects. With the recent advances in treatment techniques that have improved the survival rate, patients suffer from continuing posttreatment complications. Patients seem to cope well with the stress of treatment of BC and sustain a normal life; however, the deterioration in physical well-being makes the patient functionally inefficient. Exercise has been proven to be an effective, safe, and feasible tool in combating the adverse effects of treatment, prevents complications and decreases the risk of BC-specific mortality. This review briefly presents an overview of the burden of the disease and its management strategies. Owing to the heterogeneity of the population and the multitude of therapies they receive, the response of each patient to treatment is different and so is the magnitude of adverse effects. The review discusses the late sequelae following treatment and evidence supporting the role of physical activity in their management. In conclusion, there is a need for personalized physical activity plans to be developed to suit the individual and their circumstances.

Cytochrome P450 1B1 Contributes to the Development of Atherosclerosis and Hypertension in Apolipoprotein E-Deficient Mice.

Cytochrome P450 (CYP) 1B1 contributes to vascular smooth muscle cell growth and hypertension in male mice. This study was conducted to determine the contribution of CYP1B1 to the development of atherosclerosis and hypertension and associated pathogenesis in 8-week-old male apolipoprotein E-deficient (ApoE(-/-)/Cyp1b1(+/+)), and ApoE- and CYP1B1-deficient (ApoE(-/-)/Cyp1b1(-/-)) mice fed a normal or atherogenic diet for 12 weeks. A separate group of ApoE(-/-)/Cyp1b1(+/+) mice on an atherogenic diet was injected every third day with the CYP1B1 inhibitor, 2,3',4,5'-tetramethoxystilbene (300 µg/kg), or its vehicle, dimethyl sulfoxide (30 µL, IP); systolic blood pressure was measured by the tail cuff method. After 12 weeks, mice were euthanized, blood collected for lipid analysis, and aortas harvested for measuring lesions and remodeling, and for infiltration of inflammatory cells by histological and immunohistochemical analysis, respectively, and for reactive oxygen species production. Blood pressure, areas of lipids and collagen deposition, elastin breaks, infiltration of macrophages and T lymphocytes, reactive oxygen species generation in the aorta, and plasma lipid levels were increased in ApoE(-/-)/Cyp1b1(+/+) mice on an atherogenic diet; these changes were minimized in mice given 2,3',4,5'-tetramethoxystilbene, and in ApoE(-/-)/Cyp1b1(-/-) mice on an atherogenic diet; absorption/production of lipids remained unaltered in these mice. These data suggest that aortic lesions, hypertension, and associated pathogenesis in ApoE(-/-)/Cyp1b1(+/+) mice on an atherogenic diet are most likely dependent on CYP1B1-generated oxidative stress and increased plasma lipid levels independent of blood pressure and absorption of lipids. CYP1B1 could serve as a novel target for developing drugs to treat atherosclerosis and hypertension caused by hypercholesterolemia.

The developmental evolution of the seizure phenotype and cortical inhibition in mouse models of juvenile myoclonic epilepsy.

The GABA(A) receptor (GABA(A)R) α1 subunit mutation, A322D, causes autosomal dominant juvenile myoclonic epilepsy (JME). Previous in vitro studies demonstrated that A322D elicits α1(A322D) protein degradation and that the residual mutant protein causes a dominant-negative effect on wild type GABA(A)Rs. Here, we determined the effects of heterozygous A322D knockin (Het(α1)AD) and deletion (Het(α1)KO) on seizures, GABA(A)R expression, and motor cortex (M1) miniature inhibitory postsynaptic currents (mIPSCs) at two developmental time-points, P35 and P120. Both Het(α1)AD and Het(α1)KO mice experience absence seizures at P35 that persist at P120, but have substantially more frequent spontaneous and evoked polyspike wave discharges and myoclonic seizures at P120. Both mutant mice have increased total and synaptic α3 subunit expression at both time-points and decreased α1 subunit expression at P35, but not P120. There are proportional reductions in α3, ß2, and γ2 subunit expression between P35 and P120 in wild type and mutant mice. In M1, mutants have decreased mIPSC peak amplitudes and prolonged decay constants compared with wild type, and the Het(α1)AD mice have reduced mIPSC frequency and smaller amplitudes than Het(α1)KO mice. Wild type and mutants exhibit proportional increases in mIPSC amplitudes between P35 and P120. We conclude that Het(α1)KO and Het(α1)AD mice model the JME subsyndrome, childhood absence epilepsy persisting and evolving into JME. Both mutants alter GABA(A)R composition and motor cortex physiology in a manner expected to increase neuronal synchrony and excitability to produce seizures. However, developmental changes in M1 GABA(A)Rs do not explain the worsened phenotype at P120 in mutant mice.

Prevalence and progression of chronic kidney disease in adult patients with sickle cell disease.

AIM: We evaluated the prevalence and progression of chronic kidney disease (CKD) during the 5-year period in a cohort of patients with sickle cell disease (SCD) aged 18 years and older. METHODS: We studied 98 patients with SCD. Chronic kidney disease stages I through V were defined based on estimated glomerular filtration rate (eGFR), and albuminuria grades were defined based on spot urine protein-to-creatinine ratio according to the 2012 Kidney Disease Improving Global Outcomes recommendations. In patients with eGFR of greater than 60 mL/min per 1.73 m(2), CKD was diagnosed if grade A2 or A3 albuminuria was present. Chronic kidney disease progression was defined as an increase in CKD stage with an additional eGFR reduction of more than 25% from baseline. RESULTS: At baseline, 28.6% of patients had CKD. After a mean follow-up of 5.0 (SD, 0.9) years, 17 patients developed new CKD and the overall CKD prevalence increased to 41.8%. In addition, 8 patients experienced CKD progression. The following baseline variables were associated with the development and progression of CKD in univariate analysis: older age (P = 0.003), higher systolic blood pressure (BP; P = 0.003), lower eGFR (P = 0.001), higher serum creatinine (P = 0.001), and A3 albuminuria (P = 0.008). In multivariate analysis, baseline A3 albuminuria (adjusted odds ratio, 5.0; 95% confidence interval, 1.1-24.3; P = 0.048) and each 1-mm Hg increase in systolic BP (adjusted odds ratio, 1.04; 95% confidence interval, 1.0-1.07; P = 0.039) predicted CKD development and progression. CONCLUSIONS: Chronic kidney disease is common in patients with SCD and its prevalence increases with age. Several baseline modifiable and nonmodifiable factors were associated with the development and progression of CKD in patients with SCD. Strategies targeting BP control and proteinuria may be beneficial for individuals with SCD.