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Background: Tooth decay and traumatic dental injuries are the main causes of pulp inflammation in primary teeth. The present study was conducted on photodynamic therapy for endodontic treatment of primary teeth. Materials & Methods: 40 primary anterior teeth with pulp necrosis were divided into 2 groups: Group I, conventional root canal therapy and group II, conventional root canal therapy combined with antimicrobial PDT. Each group with 20 patients. Microbiological samples of the intra-canal content were taken using paper cones. Results: Group I & II had 19 boys and 9 girls and 8 boys and 12 girls respectively. Colony-forming units (CFU/mL) before and after treatment in group I at sample 1, 2, 3, 4, 5, 6, 7, 8, 9& 10 were 94000 and 0, 87040 and 2433, 14530 and 0,154231 and 4500, 123145 and 3200, 28453 and 550, 110232 and 1250,112456 and 3100, 43294 and 0 and 142945 and 1955 respectively. CFU before and after treatment in group II at sample 1,2,3,4,5,6,7,8,9 &10 were 194000 and 248,187040 and 2450,24530 and 0, 254231 and 4200,23140 and 300,38453 and 430,120232 and 1280, 212438 and 3600,143294 and 0 and 242940 and 1924 respectively. Conclusion: Conventional treatment combined with antimicrobial PDT found to be equally

Background: Three key elements are essential for performing REPs: stem cells, scaffolds, and activators. The present study was conducted to assess outcome of regenerative endodontics for treatment of periapical lesions. Materials & Methods: 40 patients with mature incisors, canines, or mandibular premolars showing pulp necrosis and apical periodontitis were divided into group I (REP) and conventional root canal treatment (ENDO) group II. Calcium hydroxide medication was used, and the cavity was sealed. 3 weeks later, patients were treated following their assigned protocol of ENDO or REP. Clinical follow-up examinations were performed at 6 months and 12 months. Results: At 6 months, success was seen in 20 teeth in group I and 18 in group II and failure in 2 in group II. At 12 months, success was seen in 20 in each group. The mean height (mm) at 6 months was 2.5 and 2.8 in group I and II, at 12 months was 1.6 and 1.9. Anterior-posterior dimension at 6 months was 2.6 and 2.7 and at 12 months was 1.8 and 2.2. Mesiodistal dimension at 6 months was 3.1 and 2.5 and at 12 months was 1.8 and 2.1 in group I and II respectively.

This case series describes the use of ultrasound (US)-guided dorsal sacral foraminal block (DSFB) for providing postoperative analgesia in six patients who underwent foot and ankle surgeries under spinal anesthesia. Postoperatively, all of them received a US-guided DSFB at the level of the brim of the second sacral foramina (SF2). Needle placements were confirmed with fluoroscopic (FL) images and injected radiocontrast defined the diffusion with a postoperative CT scan. The images obtained depicted ipsilateral spread in the sacral epidural space, sacral nerve roots, and plexus. The US-guided DSFB could be effectively used as an alternative method for postoperative pain relief after foot and ankle surgery.

BACKGROUNDMembers of the cysteine-rich secretory proteins (CRISPS), antigen 5 (Ag5) and pathogenesis-related 1 (Pr-1) (CAP) superfamily of proteins are found across the bacterial, fungal, plant and animal kingdoms. Although many CAP superfamily proteins remain poorly characterized, over the past decade evidence has accumulated, which provides insights into the functional roles of these proteins in various processes, including fertilization, immune defence and subversion, pathogen virulence, venom toxicology and cancer biology.OBJECTIVE AND RATIONALEThe aim of this article is to summarize the current state of knowledge on CAP superfamily proteins in mammalian fertility, organismal homeostasis and disease pathogenesis.SEARCH METHODSThe scientific literature search was undertaken via PubMed database on all articles published prior to November 2019. Search terms were based on following keywords: ‘CAP superfamily’, ‘CRISP’, ‘Cysteine-rich secretory proteins’, ‘Antigen 5’, ‘Pathogenesis-related 1’, ‘male fertility’, ‘CAP and CTL domain containing’, ‘CRISPLD1’, ‘CRISPLD2’, ‘bacterial SCP’, ‘ion channel regulator’, ‘CatSper’, ‘PI15’, ‘PI16’, ‘CLEC’, ‘PRY proteins’, ‘ASP proteins’, ‘spermatogenesis’, ‘epididymal maturation’, ‘capacitation’ and ‘snake CRISP’. In addition to that, reference lists of primary and review article were reviewed for additional relevant publications.OUTCOMESIn this review, we discuss the breadth of knowledge on CAP superfamily proteins with regards to their protein structure, biological functions and emerging significance in reproduction, health and disease. We discuss the evolution of CAP superfamily proteins from their otherwise unembellished prokaryotic predecessors into the multi-domain and neofunctionalized members found in eukaryotic organisms today. At least in part because of the rapid evolution of these proteins, many inconsistencies in nomenclature exist within the literature. As such, and in part through the use of a maximum likelihood phylogenetic analysis of the vertebrate CRISP subfamily, we have attempted to clarify this confusion, thus allowing for a comparison of orthologous protein function between species. This framework also allows the prediction of functional relevance between species based on sequence and structural conservation.WIDER IMPLICATIONSThis review generates a picture of critical roles for CAP proteins in ion channel regulation, sterol and lipid binding and protease inhibition, and as ligands involved in the induction of multiple cellular processes.

Fertilization requires sperm to travel long distances through the complex environment of the female reproductive tract. Despite the strong association between poor motility and infertility, the kinetics of sperm tail movement and the role individual proteins play in this process is poorly understood. Here, we use a high spatiotemporal sperm imaging system and an analysis protocol to define the role of CRISPs in the mechanobiology of sperm function. Each of CRISP1, CRISP2, and CRISP4 is required to optimize sperm flagellum waveform. Each plays an autonomous role in defining beat frequency, flexibility, and power dissipation. We thus posit that the expansion of the CRISP family from one member in basal vertebrates, to three in most mammals, and four in numerous rodents, represents an example of neofunctionalization wherein proteins with a common core function, boosting power output, have evolved to optimize different aspects of sperm tail performance.

Background & objectives: Due to limited information available on the frequency and spectrum of cystic fibrosis (CF) transmembrane conductance regulator (CFTR) gene mutations in congenital bilateral absence of vas deferens (CBAVD) in Indian population, it is difficult to provide accurate genetic counselling to couples. The present study was undertaken to investigate the spectrum and frequency of CFTR gene mutations in Indian men with CBAVD and to determine the female CF carrier status. Methods: Direct DNA sequencing of the CFTR gene was carried out in eighty CBAVD men, their female partners and fifty controls from the general population. Pathological significance of the identified novel CFTR gene variants was carried out using in silico tools. Appropriate genetic counselling was provided to the couples prior to intracytoplasmic sperm injection (ICSI). Results: A significant association was observed for CFTR gene variants in Indian CBAVD men versus controls (odds ratio: 12.1; 95% confidence interval: 4.8-30.4; PG and IVS1+2T>G) and ten previously reported mutations (R75Q, c.1210-12[5], F508del, A309G, R334W, I444T, R668C, R709X, A1285V and Q1352H) were detected in CBAVD men. The novel and reported CFTR gene mutations were L926W (2.5%, P=0.26), R1453Q (2.5%, P=0.26), F508del (8.75%, P=0.03) and c.1210-12[5] (42.5%, P=0.002). A total of 13 (16.2%) female partners were found to be a CF carrier. Nine couples had a risk of transmitting mutant CFTR allele to the offspring. Interpretation & conclusions: The heterogeneous spectrum of CFTR gene in Indian population suggests the necessity of screening CBAVD men and female partners for accurate genetic counselling prior to undergoing ICSI.

While much is known about the microstructure of sperm flagella, the mechanisms behind the generation of flagellar beating patterns by the axoneme are still not fully understood. We demonstrate a technique for investigating the energetics of flagella or cilia. We record the planar beating of tethered wildtype andCrisp2-knockout mouse sperm at high-speed and high-resolution and extract centerlines using digital image processing techniques. We accurately reconstruct beating waveforms using a Chebyshev-polynomial based Proper Orthogonal Decomposition of the centerline tangent-angle profiles. External hydrodynamic forces and the internal resistance from the passive flagellar material are calculated from the observed kinematics of the beating patterns using a Soft, Internally-Driven Kirchhoff-Rod (SIDKR) model. Energy conservation is employed to further compute the flagellar energetics. We thus obtain the distribution of mechanical power exerted by the dynein motors without any further assumptions about mechanisms regulating axonemal function. We find that, in both the mouse genotypes studied, a large proportion of the mechanical power exerted by the dynein motors is dissipated internally, within the passive structures of the flagellum and by the motors themselves. This internal dissipation is considerably greater than the hydrodynamic dissipation in the aqueous medium outside. The net power input from the dynein motors in sperm fromCrisp2-knockout mice is significantly smaller than in corresponding wildtype samples. The reduced power is correlated with slower beating and smaller amplitudes. These measurements of flagellar energetics indicate that the ion-channel regulating cysteine-rich secretory proteins (CRISPs) may also be involved in regulating mammalian sperm motility.

Background The sperm protein IZUMO1 (Izumo sperm-egg fusion 1) and its recently identified binding partner on the oolemma, IZUMO1R, are among the first ligand-receptor pairs shown to be essential for gamete recognition and adhesion. However, the IZUMO1-IZUMO1R interaction does not appear to be directly responsible for promoting the fusion of the gamete membranes, suggesting that this critical phase of the fertilization cascade requires the concerted action of alternative fusogenic machinery. It has therefore been proposed that IZUMO1 may play a secondary role in the organization and/or stabilization of higher-order heteromeric complexes in spermatozoa that are required for membrane fusion. Results Here, we show that fertilization-competent (acrosome reacted) mouse spermatozoa harbor several high molecular weight protein complexes, a subset of which are readily able to adhere to solubilized oolemmal proteins. At least two of these complexes contain IZUMO1 in partnership with GLI pathogenesis-related 1 like 1 (GLIPR1L1). This interaction is associated with lipid rafts and is dynamically remodeled upon the induction of acrosomal exocytosis in preparation for sperm adhesion to the oolemma. Accordingly, the selective ablation of GLIPR1L1 leads to compromised sperm function characterized by a reduced ability to undergo the acrosome reaction and a failure of IZUMO1 redistribution. Conclusions Collectively, this study characterizes multimeric protein complexes on the sperm surface and identifies GLIPRL1L1 as a physiologically relevant regulator of IZUMO1 function and the fertilization process.

The cysteine-rich secretory proteins (CRISPs) are a group of proteins that show a pronounced expression biased to the male reproductive tract. Although sperm encounter CRISPs at virtually all phases of sperm development and maturation, CRISP2 is the sole CRISP produced during spermatogenesis, wherein it is incorporated into the developing sperm head and tail. In this study we tested the necessity for CRISP2 in male fertility using Crisp2 loss-of-function mouse models. In doing so, we revealed a role for CRISP2 in establishing the ability of sperm to undergo the acrosome reaction and in establishing a normal flagellum waveform. Crisp2-deficient sperm possess a stiff midpiece and are thus unable to manifest the rapid form of progressive motility seen in wild type sperm. As a consequence, Crisp2-deficient males are subfertile. Furthermore, a yeast two-hybrid screen and immunoprecipitation studies reveal that CRISP2 can bind to the CATSPER1 subunit of the Catsper ion channel, which is necessary for normal sperm motility. Collectively, these data define CRISP2 as a determinant of male fertility and explain previous clinical associations between human CRISP2 expression and fertility.

Background & objectives: The role of cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations in congenital bilateral absence of vas deferens and unilateral renal agenesis (CBAVD-URA) has been controversial. Here, we report the cases of five Indian males with CBAVD-URA. The objective was to evaluate the presence or absence of CFTR gene mutations and variants in CBAVD-URA. The female partners of these males were also screened for cystic fibrosis (CF) carrier status. Methods: Direct DNA sequencing of CFTR gene was carried out in five Indian infertile males having CBAVD-URA. Female partners (n=5) and healthy controls (n=32) were also screened. Results: Three potential regulatory CFTR gene variants (c.1540A>G, c.2694T>G and c.4521G>A) were detected along with IVS8-5T mutation in three infertile males with CBAVD-URA. Five novel CFTR gene variants (c.621+91A>G, c.2752+106A>T, c.2751+85_88delTA, c.3120+529InsC and c.4375-69C>T), four potential regulatory CFTR gene variants (M470V, T854T, P1290P, Q1463Q) and seven previously reported CFTR gene variants (c.196+12T>C, c.875+40A>G, c.3041-71G>C, c.3271+42A>T, c.3272-93T>C, c.3500-140A>C and c.3601-65C>A) were detected in infertile men having CBAVD and renal anomalies Interpretation & conclusions: Based on our findings, we speculate that CBAVD-URA may also be attributed to CFTR gene mutations and can be considered as CFTR-related disorder (CFTR-RD). The CFTR gene mutation screening may be offered to CBAVD-URA men and their female partners undergoing ICSI. Further studies need to be done in a large sample to confirm the findings.

The stability analysis of a nuclear reactor is an important aspect in the design and operation of the reactor. A stable neutronic response to perturbations is essential from the safety point of view. In this paper, a general methodology has been developed for the linear stability analysis of nuclear reactors using the lumped reactor model. The reactor kinetics has been modelled using the point kinetics equations and the reactivity feedbacks from fuel, coolant and xenon have been modelled through the appropriate time dependent equations. These governing equations are linearized considering small perturbations in the reactor state around a steady operating point. The characteristic equation of the system is used to establish the stability zone of the reactor considering the reactivity coefficients as parameters. This methodology has been used to identify the stability region of a typical pressurized heavy water reactor. It is shown that the positive reactivity feedback from xenon narrows down the stability region. Further, it is observed that the neutron kinetics parameters (such as the number of delayed neutron precursor groups considered, the neutron generation time, the delayed neutron fractions, etc.) do not have a significant influence on the location of the stability boundary. The stability boundary is largely influenced by the parameters governing the evolution of the fuel and coolant temperature and xenon concentration.

The aim of the study was to detect the frequency of the CFTR gene variants poly-T, TG repeats and c.1408A>G p.Met470Val (M470V) in Indian men with congenital bilateral absence of the vas deferens (CBAVD). Men diagnosed with CBAVD (n = 76), their female partners (n = 76) and healthy men from general population (n = 50) were recruited. Genomic DNA was isolated and the polymorphic regions of IVS9- c.1210-12T [5] and M470V were amplified using specific primers followed by Sanger's DNA sequencing. A statistically significant increase in the frequency of heterozygous IVS9- c.1210-12T [5] (39.4%) was observed in CBAVD men as compared to controls (14%). The allelic distribution of c.1210-12T [5], c.1210-12T [7] and c.1210-12T [9] in CBAVD men was 21%, 64.4% and 13% and that in healthy controls was 7%, 73% and 20% respectively. Longest TG repeat c.1210-34TG [13] was found in association with c.1210-12T [5] with an allelic frequency of 5.9% in CBAVD men. We found a significant association of c.1210-34TG [12]/c.1210-34TG [13] - c.1210-12[5] -V470 allele in CBAVD men. Twelve female partners harboured a heterozygous c.1210-12T [5] allele. The study emphasises the need to screen both partners for the polymorphisms M470V, poly-T, TG tract repeats in addition to population-specific known CFTR gene mutations.

A simple and rapid liquid chromatography/tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the simultaneous quantitation of Olanzapine and Fluoxetine in human plasma using Olanzapine-d3 and Fluoxetine-d5 HCl as internal standard (IS), respectively. After solid phase extraction of the plasma samples on Waters Oasis HLB Catridges, Olanzapine, Fluoxetine and IS were chromatographed on Thermo Hypersil Gold C18 (50 mm × 4.6 mmi.d., 5 μm) analytical column with isocratic elution using methanol: 2mM Ammonium acetate buffer (90:10). Quantitation was performed on a triple quadrupole mass spectrometer employing electrospray ionization technique and operating in multiple reaction monitoring (MRM) and positive ion mode with transitions at 313/256 for Olanzapine and 310/148 for Fluoxetine. The total chromatographic run time was 2.0 min and calibration curves were linear over the concentration range of 0.10-20.00 ng/mL for Olanzapine and 0.50-50.00 ng/mL for Fluoxetine. The method was validated for selectivity, sensitivity, recovery, linearity, accuracy and precision and stability studies. The recoveries obtained for the Olanzapine and its IS was ≥87% and Fluoxetine and its IS was ≥91%. Recoveries obtained were consistent and reproducible. Inter-batch and intra-batch coefficient of variation across three validation runs (LLOQ, LQC, MQC1, MQC and HQC) was less than 3.6 for Olanzapine and less than 5.2% for Fluoxetine. The method was successfully applied to a pharmacokinetic study of fixed dose combination of Olanzapine/Fluoxetine in healthy male volunteers.

Objective To develop a simple, sensitive and selective LC–MS–MS method for the simultaneous determination of amoxicillin and clavulanic acid in human plasma using amoxicillin D4 and ampicillin as internal standard (IS). Method Amoxicillin and clavulanic acid in plasma were concentrated by solid phase extraction and chromatographed on a C18 column using a mobile phase of acetonitrile: 2 mM ammonium acetate (80:20 v/v) under isocratic condition. Quantitation was performed on a triple quadrupole mass spectrometer by employing electrospray ionization technique and operating in multiple reaction monitoring (MRM) and negative ion mode with mass transitions 364.00/223.00, 198.00/136.00, 368.00/227.00 and 347.90/304.00 for Amoxicillin, Clavulanic acid, Amoxicillin D4 and Ampicillin respectively. Results The method was validated over a linear range of 50.43–31500.68 and 25.28–6185.18 ng/mL for amoxicillin and clavulanic acid respectively. The Lower Limit of Quantitation (LLOQ) were 50.43 and 25.28 ng/mL for amoxicillin and clavulanic acid respectively. Inter-batch and intra-batch coefficient of variation across three validation runs (LLOQ, LQC, MQC1, MQC and HQC) was less than 3.55% and 3.07% for amoxicillin and clavulanic acid respectively. Conclusion The method was validated and was suitable for the quantitation of amoxicillin and clavulanic acid from plasma samples in a pharmacokinetic study.