Association between coronary artery vitamin D receptor expression and select systemic risks factors for coronary artery atherosclerosis.

OBJECTIVE: The aim of this study is to analyze the association between coronary artery vitamin D receptor (VDR) expression and systemic coronary artery atherosclerosis (CAA) risk factors. METHODS: Female cynomolgus monkeys (n = 39) consumed atherogenic diets containing the women's equivalent of 1000 IU/day of vitamin D3. After 32 months consuming the diets, each monkey underwent surgical menopause. After 32 postmenopausal months, CAA and VDR expression were quantified in the left anterior descending coronary artery. Plasma 25OHD3, lipid profiles and serum monocyte chemotactic protein-1 (MCP-1) were measured. RESULTS: In postmenopausal monkeys receiving atherogenic diets, serum MCP-1 was significantly elevated compared with baseline (482.2 ± 174.2 pg/ml vs. 349.1 ± 163.2 pg/ml, respectively; p < 0.001; d = 0.79) and at the start of menopause (363.4 ± 117.2 pg/ml; p < 0.001; d = 0.80). Coronary VDR expression was inversely correlated with serum MCP-1 (p = 0.042). Additionally, the change of postmenopausal MCP-1 (from baseline to necropsy) was significantly reduced in the group with higher, compared to below the median, VDR expression (p = 0.038). The combination of plasma 25OHD3 and total plasma cholesterol/high-density lipoprotein cholesterol was subsequently broken into low-risk, moderate-risk and high-risk groups; as the risk increased, the VDR quantity decreased (p = 0.04). CAA was not associated with various atherogenic diets. CONCLUSION: Coronary artery VDR expression was inversely correlated with markers of CAA risk and inflammation, including MCP-1, suggesting that systemic and perhaps local inflammation in the artery may be associated with reduced arterial VDR expression.

Development and evaluation of a novel taekwondo chest protector to improve mobility when performing axe kicks.

The axe kick, in Olympic style taekwondo, has been identified as the most popular scoring technique aimed to the head during full contact competition. The first purpose of this study was to identify and investigate design issues with the current World Taekwondo Federation approved chest protector. A secondary purpose was to develop a novel chest protector addressing the identified design issues and to conduct a biomechanical analysis. Fifteen male elite Taekwondo players were selected to perform three different styles of the axe kick, i.e., front, in-out, and out-in axe kick five times each for a total of 45 kicks. Two-way repeated measures ANOVA showed significant differences between the novel and existing chest protector conditions for vertical height of the toe, downward kicking foot speed, hip flexion angle and ipsilateral shoulder flexion extension range of motion (ROM) (p < 0.05). There were no significant differences between the control condition (no chest protector) and the novel chest protector condition for these variables (p > 0.05). These results indicate that the novel chest protector interferes less with both the lower and upper limbs during the performance of the axe kick and provides a more natural, free-moving alternative to the current equipment used.

Association between proton pump inhibitors and spontaneous bacterial peritonitis in cirrhotic patients - a systematic review and meta-analysis.

Acid suppressive therapy, in the form of proton pump inhibitor (PPI), is widely used in cirrhotic patients, often in indications which are not clearly justified. PPI facilitates enteric bacterial colonisation, overgrowth and translocation, which might predispose to spontaneous bacterial peritonitis. However, observational studies evaluating the association of PPI and SBP in cirrhotic patients have yielded inconsistent results. We therefore conducted a meta-analysis of relevant clinical studies to determine the nature of this association. Observational studies assessing the association between SBP and PPI in cirrhosis, conducted in adult population and published in all languages, were identified through systematic search in the MEDLINE, EMBASE and manual reviews of all major gastroenterology meeting proceedings up to May 2010. The relevant studies were pooled using traditional meta-analytic techniques with a random-effects model. Four studies were identified and included in the meta-analysis. The pooled analysis, involving a total of 772 patients, found a significant association between the use of PPI and the development of SBP (OR 2.77, 95% CI 1.82-4.23). There was very little degree of heterogeneity as reflected by an I(2) value of 22% and the visual inspection of the funnel plot. There is a potential association between use of PPI and development of SBP. Therefore, PPIs should be used judiciously and only when clearly indicated in cirrhotics. Further studies are essential to clarify this relationship and elucidate the underlying mechanisms.

Seroprevalence of SARS-CoV-2 antibodies, associated epidemiological factors and antibody kinetics among healthcare workers in Connecticut.

BACKGROUND: Healthcare workers (HCWs) are at the front line of the ongoing coronavirus 2019 (COVID-19) pandemic. Comprehensive evaluation of the seroprevalence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) among HCWs in a large healthcare system could help to identify the impact of epidemiological factors and the presence of symptoms on the immune response to the infection over time. AIM: To determine the seroprevalence of SARS-CoV-2-specific antibodies among HCWs, identify associated epidemiological factors and study antibody kinetics. METHODS: A longitudinal evaluation of the seroprevalence and epidemiology of SARS-CoV-2-specific antibodies was undertaken in approximately 30,000 HCWs in the largest healthcare system in Connecticut, USA. FINDINGS: At baseline, the prevalence of SARS-CoV-2 antibody among 6863 HCWs was 6.3% [95% confidence interval (CI) 5.7-6.9%], and was highest among patient care support (16.7%), medical assistants (9.1%) and nurses (8.2%), and lower for physicians (3.8%) and advanced practice providers (4.5%). Seroprevalence was significantly higher among African Americans [odds ratio (OR) 3.26 compared with Caucasians, 95% CI 1.77-5.99], in participants with at least one symptom of COVID-19 (OR 3.00, 95% CI 1.92-4.68), and in those reporting prior quarantine (OR 3.83, 95% CI 2.57-5.70). No symptoms were reported in 24% of seropositive participants. Among the 47% of participants who returned for a follow-up serological test, the seroreversion rate was 39.5% and the seroconversion rate was 2.2%. The incidence of re-infection in the seropositive group was zero. CONCLUSION: Although there is a decline in the immunoglobulin G antibody signal over time, 60.5% of seropositive HCWs had maintained their seroconversion status after a median of 5.5 months.

Four Ia invariant chain forms derive from a single gene by alternate splicing and alternate initiation of transcription/translation.

We determined the structural basis for the presence of electrophoretically-distinct, antigenically-related forms of invariant chains in Ia oligomers, and established the mechanisms by which they can be expressed from a single gene. S1 nuclease protection assays indicated that, in B cells, transcription of this gene initiates at a minimum of three sites. Thus, unlike previously thought, invariant chain mRNAs have heterogeneous 5' untranslated segments that may differentially affect initiation of translation. Further, restriction mapping and nucleotide sequencing of cDNAs revealed two kinds of invariant chain mRNAs differing by an internal coding segment of 192 bp. This segment represents an alternatively spliced exon, as demonstrated by nucleotide sequencing of corresponding genomic regions. The exon (exon X) encodes a cysteine-rich stretch of 64 amino acids near the COOH terminus that displays a striking and surprising homology to an internal amino acid repeat of thyroglobulin, suggesting an evolutionary mechanism of exon shuffling. Transient expression of cDNAs indicated that both types of alternatively spliced mRNAs contain two in-frame AUGs functioning as alternate start sites for translation. Thus, transfections with exon X-lacking cDNAs resulted in the expression of Mr 33,000 and 31,000 proteins, detected by immunoprecipitation with anti-invariant chain antisera, and identical by two-dimensional gel (2-D) analyses to the B cell invariant-chain forms gamma 1 (Mr 31,000), gamma 2, and gamma 3 (Mr 33,000). Similarly, exon X-containing cDNAs expressed Mr 43,000 and 41,000 proteins, also identical by 2-D migration to Ia-associated proteins. Thus, human Ia molecules contain four forms of invariant chain of closely related but nonidentical primary structure that are generated from a single gene by a complex pattern of alternate transcriptional start, exon splicing, and translational start.

The human invariant chain is the core protein of the human class II-associated proteoglycan.

The human class II-associated chondroitin sulfate proteoglycan (CSPG) was analyzed biochemically and immunologically to determine a possible relationship with the human invariant chain (gamma 1) and its related components. The CSPG was purified by a three-step procedure involving associative ion-exchange chromatography, immunoprecipitation, and dissociative ion-exchange chromatography. Treatment of the CSPG with chondroitinase revealed core proteins of Mr approximately 46,000, 38,000, and 28,000, with the 38,000 species most highly represented. Tryptic peptide analysis revealed identity of the peptides of the 38,000 Mr core protein and gamma 1, and of the 28,000 Mr species and p25. The CSPG and its core proteins were shown to react directly with the mouse anti-human invariant chain monoclonal antibody VIC-Y1 and a rabbit antiserum produced against a synthetic peptide corresponding to the C-terminal end of invariant chain. These results demonstrate that the invariant chain is the core protein of the class II-associated CSPG. In addition, virtually all the CSPG was shown to be present on the cell surface.

The effect of oxidative stress on the mutation rate of Mycobacterium tuberculosis with impaired catalase/peroxidase function.

OBJECTIVES: To determine the effect of oxidative stress on isoniazid-resistant Mycobacterium tuberculosis deficient in catalase/peroxidase activity to varying degrees through mutation in katG. METHODS: The mutation rate was determined for a set of isogenic strains with different katG alleles giving different catalase and/or peroxidase activities following exposure to the oxidizing agent, hydrogen peroxide. Mutants were selected on rifampicin, and the location and nature of the mutation were identified by sequencing the rpoB gene. RESULTS: No evidence was found to suggest that strains that had impaired catalase/peroxidase activity were hypermutable, and the presence of excess hydrogen peroxide had no effect on the mutation rate. An unusual pattern of mutations in rpoB was observed in catalase-deficient strains with only 3 of 66 having mutations within the rifampicin resistance-determining region. CONCLUSIONS: The mutation rate of M. tuberculosis in response to oxidative stress is not increased in strains with significant deficits in catalase and peroxidase activity. Our data suggest that isoniazid-resistant strains compensate for their reduced ability to detoxify oxidative stress effectively. Interestingly, mutations were found in unusual locations at positions similar to those found in clinical isoniazid-resistant strains.

Mycobacterium tuberculosis DNA repair in response to subinhibitory concentrations of ciprofloxacin.

OBJECTIVES: To investigate how the SOS response, an error-prone DNA repair pathway, is expressed following subinhibitory quinolone treatment of Mycobacterium tuberculosis. METHODS: Genome-wide expression profiling followed by quantitative RT (qRT)-PCR was used to study the effect of ciprofloxacin on M. tuberculosis gene expression. RESULTS: Microarray analysis showed that 16/110 genes involved in DNA protection, repair and recombination were up-regulated. There appeared to be a lack of downstream genes involved in the SOS response. qRT-PCR detected an induction of lexA and recA after 4 h and of dnaE2 after 24 h of subinhibitory treatment. CONCLUSIONS: The pattern of gene expression observed following subinhibitory quinolone treatment differed from that induced after other DNA-damaging agents (e.g. mitomycin C). The expression of the DnaE2 polymerase response was significantly delayed following subinhibitory quinolone exposure.

Technique for cloning and sequencing the ends of bacterial artificial chromosome inserts.

Bacterial artificial chromosome (BAC) libraries are an important tool for positional cloning, gene analysis and physical mapping. During studies using BAC clones, it is often necessary to organize them into contiguous sequences (contigs). To finalize, join and extend the contigs, both cloning and sequencing of the ends of the inserts are required. Here, we describe a low-cost, accessible, fast and powerful method for the routine isolation of BAC ends. This method allows the isolation of 20 BAC clone ends in one day. The analysis of the ends reveals fragment sizes compatible with sequencing, and the structure of these clones allows the sequencing of both ends using the same plasmid. Moreover, long end fragments can be sequenced in both directions.

The HLA-class II-associated chondroitin sulfate proteoglycan expressed by class II positive T and monocyte-like cell lines is larger than that expressed by EBV-transformed B-lymphoblastoid cell lines.

The human class II-associated chondroitin sulfate proteoglycan (CSPG) was originally detected as an approximately 40-70 kd species from normal human tonsil cells and Epstein-Barr virus (EBV) transformed B-lymphoblastoid cell lines. The identification of the invariant chain as the core protein of the CSPG allowed us to directly assay for the CSPG on both class II positive and negative immunocompetent cells of other lineages. Our results indicate that the CSPG is present on the class II-positive monocyte-like cell line U937 and T-cell line HUT-102, but not on the class-II negative T-cell line CCRF/CEM. No class II positive cells were found that did not also express the CSPG. The expression of the CSPG on U937 cells is increased after stimulation with gamma-interferon and PMA, paralleling the previously described increase in class II and invariant chain expression. In addition, the CSPGs from U937, HUT-102, and Raji, all cell lines derived from human malignancies, migrate as an approximately 55-90 kd species, larger than the CSPG previously characterized. However, the core proteins of the CSPG from all cells studied appear as two bands of 38 and 28 kd, indicating the size difference in the CSPG is attributable to differences in the glycosaminoglycan chains.

Maxillary anterior caries associated with increased caries risk in other primary teeth.

It has been suggested that children with caries of maxillary anterior primary teeth may have increased caries incidence in other teeth. This study aimed to quantify the extent of posterior dental caries in those children who initially presented with maxillary anterior caries compared with that of those who did not. Data were collected for 217 Head Start children participating in a one-year study to determine caries risk factors. Tooth surfaces were recorded as carious, restored, or extracted (missing due to caries). Caries was then categorized into the following pattern(s): maxillary anterior (MA), pit/fissure (PF), posterior proximal (PP), and posterior buccal/lingual smooth surface (BL). Compared with children who did not initially present with the maxillary anterior pattern, those with the maxillary anterior pattern had 2.5 times (p < 0.01) the mean number of posterior decayed, missing, and filled surfaces (dmfs) and approximately 3 times the prevalence (p < 0.01) of the PP and BL patterns. The positive predictive value of the MA pattern was 86.8% for children who developed the PF pattern, and the negative predictive values were greater than 91% for children who did not develop the PP and BL patterns. The prevalence of maxillary anterior caries among children in this study is associated with a significantly greater caries incidence in posterior teeth.

Dental caries prevalence and treatment levels in Arizona preschool children.

OBJECTIVES: To assess the prevalence of dental caries in a large group of preschool children, to determine the extent to which the children received dental treatment, to examine the association between demographic and socioeconomic factors and the prevalence of caries, and to compare these findings with those from previous studies of preschool populations in the United States. METHODS: Dental caries exams were performed on 5171 children ages 5 months through 4 years, and a parent or other caregiver was asked to complete a questionnaire giving information about the child and her or his household. The children were recruited from Head Start programs; Women, Infants, and Children (WIC) nutrition programs; health fairs; and day care centers in a representative sample of Arizona communities with populations of more than 1000 people. RESULTS: Of the 994 one-year-old children examined, 6.4% had caries, with a mean dmft (decayed, missing [extracted due to caries], and filled teeth) score of 0.18. Nearly 20% of the 2-year-olds had caries, with a mean dmft of 0.70. Thirty-five percent of the 3-year-olds had caries, with a mean dmft of 1.35, and 49% of the 4-year-olds had caries, with a mean dmft of 2.36. Children whose caregivers fell into the lowest education category had a mean dmft score three times higher than those with caregivers in the highest education category. Children with caregivers in the lowest income category had a mean dmft score four times higher than those with caregivers in the highest category. Children younger than age 3 had little evidence of dental treatment, and most of the children with caries in each age group had no filled or extracted teeth. CONCLUSIONS: The data show that dental caries is highly prevalent in this preschool population, with little of the disease being treated. Timing of diagnostic examinations and prevention strategies for preschool children need to be reconsidered, especially for children identified as having a high risk of caries.

Salivary mutans streptococci and dental caries patterns in pre-school children.

Recent studies have suggested that the identification of caries as discrete patterns may be valuable in describing and predicting caries experience on an individual basis. The purpose of this study was to assess the association between levels of salivary mutans streptococci and the prevalence, incidence and distribution of caries patterns in the primary dentition. A cohort of pre-school children (n = 146, mean age 3.8 yr) were examined for dental caries and sampled for salivary mutans streptococci (SMS) at baseline and once annually for 2 yr. Children's tooth surfaces were categorized into four patterns: pit/fissure, maxillary anterior, posterior proximal, and buccal/lingual smooth surface. Salivary mutans streptococci were enumerated using a tongue blade technique, and were categorized as low (0 CFU), moderate (1-50 CFU) and high (> 50 CFU). At year 2, children with high baseline SMS had the 1) highest prevalence of caries (87%) and the highest dmfs (9.15); 2) highest prevalence of each pattern, and 3) greatest number of patterns. Among children with the pit/fissure pattern, those with high baseline SMS had the greatest pit/fissure dmfs after 2 yr. Results show that baseline SMS levels were associated with both cross-sectional and longitudinal caries experience, numbers of caries patterns, and the prevalence and severity of those patterns.

Salivary mutans streptococci and caries development in the primary and mixed dentitions of children.

OBJECTIVES: For more than 25 years, both cross-sectional and longitudinal studies of dental caries have focused on the role of salivary mutans streptococci (SMS) as a predictor of caries risk. This study examined the relationship between SMS and longitudinal caries development in the primary and mixed dentitions. METHODS: Eighty-five children, initial mean age 3.8 years, were examined for dental caries at baseline and once annually for 6 years. Children's SMS were sampled with a tongue blade, which was impressed onto plates containing a medium selective for SMS. After incubation, colony forming units of SMS were determined semi-quantitatively and categorized as low, moderate or high. RESULTS: Children classified as high caries risk at baseline had significantly greater (P<0.05) dmfs scores for all teeth, and in the primary molars, than children classified as moderate or low caries risk at every age but 9 (P<0.10). Children classified as high risk at age 3 had the greatest DMFS increment through age 8. Based on annual examinations, there was a trend towards increasing mean dmfs/DMFS scores among children classified as high risk in every year. CONCLUSIONS: The current study is among the first to report on the ability of annual measurements of SMS to identify long-term caries risk in both the primary and the mixed dentitions. Despite limitations in predicting caries risk using microbiological methods, this longitudinal study supports the overall benefits of this type of testing.

Mutans streptococci and caries prevalence in preschool children.

Numerous studies have reported a correlation between mutants streptococci levels and dental caries. The aim of this study was to assess the relationship between salivary mutans streptococci levels and caries in preschool children of low socioeconomic status. A total of 462 Head Start children, mean age 3.8 yr (range 2.0-5.3 yr), were examined by the modified method of Radike. Saliva samples from 458 of these children were collected with tongue blades and impressed onto mutants streptococci selective agar. Children's mutants streptococci levels were categorized as low (0 CFU), moderate (1-50 CFU) or high (> 50 CFU), and the mean dmfs was 0.40, 1.92 and 4.88, respectively. All study groups (Black, Hispanic and White) had infection rates of approximately 83%; however, 39.1% of Black children had high mutans streptococci levels compared with 28.4% of White children. Pit/fissure caries was the most prevalent disease type in children with moderate or high mutants streptococci levels, although White children in the high group had significantly less of this pattern than Blacks and Hispanics. Sensitivity, specificity, and positive and negative predictive values for the high mutants streptococci group were 91.3%, 57.5%, 69.3% and 86.3%. Results from this study indicate that differences between Black, Hispanic and White preschool children may influence caries activity within populations that have similar mutants streptococci infection levels and socioeconomic backgrounds.

The relationship between bottle usage/content, age, and number of teeth with mutans streptococci colonization in 6-24-month-old children.

OBJECTIVES: Mutans streptococci (MS) are the primary pathogens involved in the development of early childhood caries. However, factors that may affect their acquisition in the mouths of young children are not well understood, and the period of initial colonization remains controversial. This study investigated the relationship of age, number of teeth, and bottle usage/content with regard to the isolation of MS in 6-24-month-old children. METHODS: A total of 122 children from low-income families attending a nutritional supplement program, and their mothers, participated in this study. Children were examined for dental caries and number of erupted teeth and were sampled for MS. Mothers were administered a questionnaire to obtain details of baby bottle use, including what food items were put in the bottle during the last week. RESULTS: MS was detected in more than one-third of the 6-24-month-olds. Unlike some studies that suggest a later period of infectivity, approximately 20% of children under 14 months of age, including 4 of 22 infants aged 6-9 months, were colonized with MS. When examined separately, age, number of teeth, and bottle usage/content were each found to be related to the presence of MS. Mutans streptococci colonization was more likely with increasing age and number of teeth, and children whose bottles contained sweetened beverages were more likely to be colonized than children whose bottles contained milk. Logistic regression models that controlled for both age and number of teeth indicated that children who consumed sweetened beverages in their baby bottle had a statistically significant, four-fold increase in the odds of colonization by MS relative to children who consumed milk. CONCLUSIONS: The finding that approximately 20% of the children under 14 months of age were infected with MS indicates that colonization in this sample of low-income preschool children may begin earlier than suggested by some investigations. Additionally, the risk of MS colonization appears lower among infants who consume milk rather than sweetened beverages in the bottle.

The association of early dental caries patterns with caries incidence in preschool children.

OBJECTIVES: This study assessed the development of caries in preschool children over two years according to baseline caries pattern. METHODS: Connecticut Head Start children (n = 142, mean age = 3.8 years) were examined for dental caries at baseline (spring 1991) and once annually for two years. Children were categorized at baseline as caries-free, having pit and fissure (PF) caries, or having maxillary anterior (MA) caries. RESULTS: After two years, children who presented at baseline with MA or PF caries had a mean posterior dmfs of greater than seven and four times, respectively, that of children who were caries-free at baseline. When dental caries of the primary dentition was categorized by specific posterior patterns (i.e., posterior proximal [PP] and buccal/lingual [BL]), change in dmfs for the PP and BL patterns in the group that presented with pit/fissure caries at baseline were nearly four and three times greater, respectively, than for those in the caries-free group. The group that presented with maxillary anterior caries at baseline had PP and BL caries increments eight times those of children who began caries-free. CONCLUSION: Dental caries presentation in 3- to 4-year-old children can identify those children and tooth surfaces that will be at the greatest risk for future caries development.