The Latest Developments in Using Proteomic Biomarkers from Urine and Serum for Non-Invasive Disease Diagnosis and Prognosis.

Due to diagnostic improvements, medical diagnostics is demanding non-invasive or minimally invasive methods. Non-invasively obtained body fluids (eg., Urine, serum) can replace cerebral fluid, amniotic fluid, synovial fluid, bronchoalveolar lavage fluid, and others for diagnostic reasons. Many illnesses are induced by perturbations of cellular signaling pathways and associated pathway networks as a result of genetic abnormalities. These disturbances are represented by a shift in the protein composition of the fluids surrounding the tissues and organs that is, tissue interstitial fluid (TIF). These variant proteins may serve as diagnostic "signatures" for a variety of disorders. This review provides a concise summary of urine and serum biomarkers that may be used for the diagnosis and prognosis of a variety of disorders, including cancer, brain diseases, kidney diseases, and other system diseases. The studies reviewed in this article suggest that serum and urine biomarkers of various illnesses may be therapeutically useful for future diagnostics. Correct illness management is crucial for disease prognosis, hence non-invasive serum and urine biomarkers have been extensively studied for diagnosis, subclassification, monitoring disease activity, and predicting treatment results and consequences.

Factors Affecting the Usage of Wearable Device Technology for Healthcare among Indian Adults: A Cross-Sectional Study.

BACKGROUND: Wearable device technology has recently been involved in the healthcare industry substantially. India is the world's third largest market for wearable devices and is projected to expand at a compound annual growth rate of ~26.33%. However, there is a paucity of literature analyzing the factors determining the acceptance of wearable healthcare device technology among low-middle-income countries. METHODS: This cross-sectional, web-based survey aims to analyze the perceptions affecting the adoption and usage of wearable devices among the Indian population aged 16 years and above. RESULTS: A total of 495 responses were obtained. In all, 50.3% were aged between 25-50 years and 51.3% belonged to the lower-income group. While 62.2% of the participants reported using wearable devices for managing their health, 29.3% were using them daily. technology and task fitness (TTF) showed a significant positive correlation with connectivity (r = 0.716), health care (r = 0.780), communication (r = 0.637), infotainment (r = 0.598), perceived usefulness (PU) (r = 0.792), and perceived ease of use (PEOU) (r = 0.800). Behavioral intention (BI) to use wearable devices positively correlated with PEOU (r = 0.644) and PU (r = 0.711). All factors affecting the use of wearable devices studied had higher mean scores among participants who were already using wearable devices. Male respondents had significantly higher mean scores for BI (p = 0.034) and PEOU (p = 0.009). Respondents older than 25 years of age had higher mean scores for BI (p = 0.027) and Infotainment (p = 0.032). CONCLUSIONS: This study found a significant correlation with the adoption and acceptance of wearable devices for healthcare management in the Indian context.

Comparative Transcriptomic and Molecular Pathway Analyses of HL-CZ Human Pro-Monocytic Cells Expressing SARS-CoV-2 Spike S1, S2, NP, NSP15 and NSP16 Genes.

The ongoing COVID-19 pandemic is a clear and present threat to global public health. Research into how the causative SARS-CoV-2 virus together with its individual constituent genes and proteins interact with target host cells can facilitate the development of improved strategies to manage the acute and long-term complications of COVID-19. In this study, to better understand the biological roles of critical SARS-CoV-2 proteins, we determined and compared the host transcriptomic responses of the HL-CZ human pro-monocytic cell line upon transfection with key viral genes encoding the spike S1 subunit, S2 subunit, nucleocapsid protein (NP), NSP15 (endoribonuclease), and NSP16 (2'-O-ribose-methyltransferase). RNA sequencing followed by gene set enrichment analysis and other bioinformatics tools revealed that host genes associated with topologically incorrect protein, virus receptor activity, heat shock protein binding, endoplasmic reticulum stress, antigen processing and presentation were up-regulated in the presence of viral spike S1 expression. With spike S2 expression, pro-monocytic genes associated with the interferon-gamma-mediated signaling pathway, regulation of phosphatidylinositol 3-kinase activity, adipocytokine signaling pathway, and insulin signaling pathway were down-regulated, whereas those associated with cytokine-mediated signaling were up-regulated. The expression of NSP15 induced the up-regulation of genes associated with neutrophil degranulation, neutrophil-mediated immunity, oxidative phosphorylation, prion disease, and pathways of neurodegeneration. The expression of NSP16 resulted in the down-regulation of genes associated with S-adenosylmethionine-dependent methyltransferase activity. The expression of NP down-regulated genes associated with positive regulation of neurogenesis, nervous system development, and heart development. Taken together, the complex transcriptomic alterations arising from these viral-host gene interactions offer useful insights into host genes and their pathways that potentially contribute to SARS-CoV-2 pathogenesis.

SARS coronavirus outbreaks past and present-a comparative analysis of SARS-CoV-2 and its predecessors.

The Coronavirus Disease 2019 (COVID-19), a pneumonic disease caused by the SARS Coronavirus 2 (SARS-CoV-2), is the 7th Coronavirus to have successfully infected and caused an outbreak in humans. Genome comparisons have shown that previous isolates, the SARS-related coronavirus (SARSr-CoV), including the SARS-CoV are closely related, yet different in disease manifestation. Several explanations were suggested for the undetermined origin of SARS-CoV-2, in particular, bats, avian and Malayan pangolins as reservoir hosts, owing to the high genetic similarity. The general morphology and structure of all these viral isolates overlap with analogous disease symptoms such as fever, dry cough, fatigue, dyspnoea and headache, very similar to the current SARS-CoV-2. Chest CT scans for SARS-CoV-2, SARS-CoV and MERS-CoV reveal pulmonary lesions, bilateral ground-glass opacities, and segmental consolidation in the lungs, a common pathological trait. With greatly overlapping similarities among the previous coronavirus, the SARS-CoV, it becomes interesting to observe marked differences in disease severity of the SARS-CoV-2 thereby imparting it the ability to rapidly transmit, exhibit greater stability, bypass innate host defences, and increasingly adapt to their new host thereby resulting in the current pandemic. The most recent B.1.1.7, B.1.351 and P.1 variants of SARS-CoV-2, highlight the fact that changes in amino acids in the Spike protein can contribute to enhanced infection and transmission efficiency. This review covers a comparative analysis of previous coronavirus outbreaks and highlights the differences and similarities among different coronaviruses, including the most recent isolates that have evolved to become easily transmissible with higher replication efficiency in humans.

The Immunomodulatory CEA Cell Adhesion Molecule 6 (CEACAM6/CD66c) Is a Protein Receptor for the Influenza a Virus.

To establish a productive infection in host cells, viruses often use one or multiple host membrane glycoproteins as their receptors. For Influenza A virus (IAV) such a glycoprotein receptor has not been described, to date. Here we show that IAV is using the host membrane glycoprotein CD66c as a receptor for entry into human epithelial lung cells. Neuraminidase (NA), a viral spike protein, binds to CD66c on the cell surface during IAV entry into the host cells. Lung cells overexpressing CD66c showed an increase in virus binding and subsequent entry into the cell. Upon comparison, CD66c demonstrated higher binding capacity than other membrane glycoproteins (EGFR and DC-SIGN) reported earlier to facilitate IAV entry into host cells. siRNA mediated knockdown of CD66c from lung cells inhibited virus binding on cell surface and entry into cells. Blocking CD66c by antibody on the cell surface resulted in decreased virus entry. We found that CD66c is a specific glycoprotein receptor for influenza A virus that did not affect entry of non-IAV RNA virus (Hepatitis C virus). Finally, IAV pre-incubated with recombinant CD66c protein when administered intranasally in mice showed decreased cytopathic effects in mice lungs. This publication is the first to report CD66c (Carcinoembryonic cell adhesion molecule 6 or CEACAM6) as a glycoprotein receptor for Influenza A virus.

COVID-19: A Review on the Novel Coronavirus Disease Evolution, Transmission, Detection, Control and Prevention.

Three major outbreaks of the coronavirus, a zoonotic virus known to cause respiratory disease, have been reported since 2002, including SARS-CoV, MERS-CoV and the most recent 2019-nCoV, or more recently known as SARS-CoV-2. Bats are known to be the primary animal reservoir for coronaviruses. However, in the past few decades, the virus has been able to mutate and adapt to infect humans, resulting in an animal-to-human species barrier jump. The emergence of a novel coronavirus poses a serious global public health threat and possibly carries the potential of causing a major pandemic outbreak in the naïve human population. The recent outbreak of COVID-19, the disease caused by SARS-CoV-2, in Wuhan, Hubei Province, China has infected over 36.5 million individuals and claimed over one million lives worldwide, as of 8 October 2020. The novel virus is rapidly spreading across China and has been transmitted to 213 other countries/territories across the globe. Researchers have reported that the virus is constantly evolving and spreading through asymptomatic carriers, further suggesting a high global health threat. To this end, current up-to-date information on the coronavirus evolution and SARS-CoV-2 modes of transmission, detection techniques and current control and prevention strategies are summarized in this review.

Combination between antibacterial and antifungal antibiotics with phytocompounds of Artemisia annua L: A strategy to control drug resistance pathogens.

ETHNOPHARMACOLOGICAL RELEVANCE: Artemisia annua L. is a traditional Chinese medicine used for the treatment of malaria, jaundice and intense fever. AIM OF THE STUDY: The aim of the present study was to investigate the phytochemicals, antioxidants, antimicrobial and synergistic potential of methanolic and petroleum ether extracts of A. annua against bacterial and fungal pathogens. METHOD: Antioxidant activity of different concentrations of methanolic and petroleum ether extracts of A. annua was determined by DPPH free radical scavenging assay. Antimicrobial activity was determined by agar well diffusion, whereas MIC and synergistic activity was done by broth dilution method.TLC and GC-MS were done to identify active phytocompounds present in methanolic and petroleum ether extracts. RESULTS: Methanolic extract of A. annua showed higher antioxidant potential (IC50 37 0.75 ± 0.34 µg ml-1) as compared to petroleum ether extract. In antimicrobial analysis, methanolic and petroleum ether extracts of A. annua produced potent inhibitory activity against Candida strains as compared to bacterial strains. Methanolic and petroleum ether extracts of A. annua produced synergistic potential with decrease in MIC from 4 to 264 folds against bacterial (S. aureus and E. coli) and Candida strains in combination with antibacterial and antifungal antibiotics. Sub fraction I of methanolic and petroleum ether extracts was isolated through silica TLC and showed 10-fold more antimicrobial activity as compared to crude extract. GC-MS analysis of sub-fraction I of A. annua revealed 13 major phytocompounds with area more than 1%. Interestingly, 2-Propenoic acid and ridecyl ester (25.88%) were the major phytocompounds. CONCLUSION: Phytocompounds of A. annua can be used as bioenhancer of antibacterial and antifungal agents to control drug resistance.

Influenza A Virus Nucleoprotein Activates the JNK Stress-Signaling Pathway for Viral Replication by Sequestering Host Filamin A Protein.

Influenza A virus (IAV) poses a major threat to global public health and is known to employ various strategies to usurp the host machinery for survival. Due to its fast-evolving nature, IAVs tend to escape the effect of available drugs and vaccines thus, prompting the development of novel antiviral strategies. High-throughput mass spectrometric screen of host-IAV interacting partners revealed host Filamin A (FLNA), an actin-binding protein involved in regulating multiple signaling pathways, as an interaction partner of IAV nucleoprotein (NP). In this study, we found that the IAV NP interrupts host FLNA-TRAF2 interaction by interacting with FLNA thus, resulting in increased levels of free, displaced TRAF2 molecules available for TRAF2-ASK1 mediated JNK pathway activation, a pathway critical to maintaining efficient viral replication. In addition, siRNA-mediated FLNA silencing was found to promote IAV replication (87% increase) while FLNA-overexpression impaired IAV replication (65% decrease). IAV NP was observed to be a crucial viral factor required to attain FLNA mRNA and protein attenuation post-IAV infection for efficient viral replication. Our results reveal FLNA to be a host factor with antiviral potential hitherto unknown to be involved in the IAV replication cycle thus, opening new possibilities of FLNA-NP interaction as a candidate anti-influenza drug development target.

Is tetherin a true antiviral: The influenza a virus controversy.

Tetherin, an interferon-inducible gene was first discovered to be an antiviral factor in 2008. A vast range of viruses, such as influenza A virus (IAV), dengue virus, Ebola virus, HIV, and RSV, have been reported to be susceptible to the antiviral activity of tetherin. Multiple reports have been published encompassing the role of tetherin in the IAV life cycle. To date, nine reports have been published regarding the role of tetherin in the IAV life cycle, with four reports supporting tetherin as an antiviral factor while five other reports suggesting no effect. To this end, this review summarizes the list of viruses currently known to be inhibited by tetherin and describes mechanisms used by viruses to overcome the antiviral potential of tetherin. Further, using IAV as disease model, we provide existing evidence in favor and against tetherin being considered as an antiviral candidate. Subsequent analysis of the experimental procedures across IAV-tetherin published reports revealed that the experimental setup (ie, cell lines, transfection reagents, and multiplicity of infection), strain-specific activity of NS1, and differing roles of NS1 in different cell lines may add up to the contributing factors leading to the discrepancies observed.

OBJECTIVE AND SUBJECTIVE EVALUATION OF ADOLESCENT'S ORTHODONTIC TREATMENT NEEDS AND THEIR IMPACT ON SELF-ESTEEM. / AVALIAÇÃO OBJETIVA E SUBJETIVA DA NECESSIDADE DE TRATAMENTO ORTODÔNTICO DO ADOLESCENTE E SEU IMPACTO SOBRE A AUTOESTIMA.

OBJECTIVE: To investigate the presence of association between objective and subjective evaluation of orthodontic treatment needs in adolescents and their impact on their self-esteem. METHODS: Cross-sectional study with adolescents aged 10-17 years old in Sri Ganganagar city, Rajasthan, India. The objective index of orthodontic treatment need (IOTN) dental health component (DHC) and the subjective aesthetic component (AC) were used to determine the normative and the self-perception need for orthodontic treatment, respectively. The selected students were further examined for dental trauma, tooth loss, and dental caries. Rosenberg Self-Esteem Scale was applied for self-esteem level determination. Linear regression analysis was executed to test the individual association of different independent clinical variables with self-esteem scores. RESULTS: Among 1,140 studied adolescents, the prevalence of dental normative orthodontic treatment need was in 56.9% of individuals, whereas 53.3% of individuals considered themselves as needy for the treatment. Multivariate analyses revealed that out of all dental disorders, DHC followed by AC of IOTN had maximum impact on the self-esteem of the adolescence. CONCLUSIONS: Dissatisfaction with dental appearance is a strong predictor for low self-esteem in adolescence.

OBJETIVO: Verificar a presença de associação entre a avaliação objetiva e subjetiva da necessidade de tratamento ortodôntico em adolescentes e seu impacto na autoestima. MÉTODOS: Estudo transversal com adolescentes de 10 a 17 anos da cidade de Sri Ganganagar, Rajastão, Índia. Foi utilizado o Índice de Ortodontia de Saúde Dental (IOTN) para avaliar de modo objetivo a necessidade de tratamento (DHC) e o componente estético desse índice (AC) para a avaliação subjetiva. Os estudantes selecionados foram também avaliados quanto a traumatismos dentários, perda dentária e cárie dentária. Aplicou-se a escala de autoestima de Rosenberg para determinar o nível de autoestima dos estudantes. Os dados foram analisados por regressão linear múltipla, testando a associação individual das diferentes variáveis clínicas independentemente da autoestima dos adolescentes, de acordo com o escore da escala de Rosenberg. RESULTADOS: Entre 1.140 adolescentes estudados, a prevalência da necessidade de tratamento ortodôntico avaliado de modo objetivo pelo IOTN-DHC ocorreu em 56,9% dos indivíduos. A avaliação subjetiva do adolescente (IOTN-AC) revelou prevalência de necessidade de tratamento de 53,3%. A análise multivariada mostrou que, além de todos os problemas dentários, o componente objetivo (IOTN-DHC), seguido do componente estético subjetivo (IOTN-AC), teve maior impacto na autoestima dos estudantes analisados. CONCLUSÕES: A insatisfação com a aparência dental é um forte preditor de baixa autoestima na adolescência.

Traumatic Dental Injuries Prevalence and their Impact on Self-esteem among Adolescents in India: A Comparative Study.

INTRODUCTION: Adolescents are mostly injured during sport activities, traffic accidents and some forms of violence which may lead to traumatic injuries. Traumatic injuries not only pose a health risk worldwide but are also regarded among serious social problems. Poor oral health has been related to poor social relationships that affect the ability to learn and grow which may lead to reduced self-esteem. AIM: This study was conducted to compare self-esteem of adolescents with and without anterior Traumatic Dental Injury (TDI). MATERIALS AND METHODS: A population based comparative study was conducted among 10 to 17 years old adolescents from November 2014 to January 2016. The study sample comprised of 424 controls and 212 cases with 2:1 control-to-case ratio that were selected by four examiners along with a gold standard examiner. TDI was recorded using Ellis classification and self-esteem was recorded using Rosenberg's Self-Esteem Scale (RSES). For comparing self-esteem of cases and controls according to RSES questions, t-test is used. Odds ratio (OR) was utilized to assess and compare the self-esteem according to high and low esteem in cases and controls. RESULTS: Ellis Class I, Ellis Class II and Ellis Class III fractures were observed in 53.3%, 33.49% and 13.20% of cases, respectively. The maxillary arch was the most affected (72.48%) and on the hierarchy of causes, this study found that on most occasions dental injuries were caused by sports (40.09%). The total mean score of RSES in cases (23.16) and in controls (24.43) was also statistically significant (p<0.001). CONCLUSION: Improving oral health in schools may contribute considerably to increase the self-esteem of adolescents in a positive direction and help them in becoming socially competent adults.

Impact of Dental Disorders and its Influence on Self Esteem Levels among Adolescents.

INTRODUCTION: Self esteem is more of a psychological concept therefore, even the common dental disorders like dental trauma, tooth loss and untreated carious lesions may affect the self esteem thus influencing the quality of life. AIM: This study aims to assess the impact of dental disorders among the adolescents on their self esteem level. MATERIALS AND METHODS: The present cross-sectional study was conducted among 10 to 17 years adolescents. In order to obtain a representative sample, multistage sampling technique was used and sample was selected based on Probability Proportional to Enrolment size (PPE). Oral health assessment was carried out using WHO type III examination and self esteem was estimated using the Rosenberg Self Esteem Scale score (RSES). The descriptive and inferential analysis of the data was done by using IBM SPSS software. Logistic and linear regression analysis was executed to test the individual association of different independent clinical variables with self esteem. RESULTS: Total sample of 1140 adolescents with mean age of 14.95 ±2.08 and RSES of 27.09 ±3.12 were considered. Stepwise multiple linear regression analysis was applied and best predictors in relation to RSES in the descending order were Dental Health Component (DHC), Aesthetic Component (AC), dental decay {(aesthetic zone), (masticatory zone)}, tooth loss {(aesthetic zone), (masticatory zone)} and anterior fracture of tooth. CONCLUSION: It was found that various dental disorders like malocclusion, anterior traumatic tooth, tooth loss and untreated decay causes a profound impact on aesthetics and psychosocial behaviour of adolescents, thus affecting their self esteem.

AVALIAÇÃO OBJETIVA E SUBJETIVA DA NECESSIDADE DE TRATAMENTO ORTODÔNTICO DO ADOLESCENTE E SEU IMPACTO SOBRE A AUTOESTIMA / OBJECTIVE AND SUBJECTIVE EVALUATION OF ADOLESCENT'S ORTHODONTIC TREATMENT NEEDS AND THEIR IMPACT ON SELF-ESTEEM

RESUMO Objetivo: Verificar a presença de associação entre a avaliação objetiva e subjetiva da necessidade de tratamento ortodôntico em adolescentes e seu impacto na autoestima. Métodos: Estudo transversal com adolescentes de 10 a 17 anos da cidade de Sri Ganganagar, Rajastão, Índia. Foi utilizado o Índice de Ortodontia de Saúde Dental (IOTN) para avaliar de modo objetivo a necessidade de tratamento (DHC) e o componente estético desse índice (AC) para a avaliação subjetiva. Os estudantes selecionados foram também avaliados quanto a traumatismos dentários, perda dentária e cárie dentária. Aplicou-se a escala de autoestima de Rosenberg para determinar o nível de autoestima dos estudantes. Os dados foram analisados por regressão linear múltipla, testando a associação individual das diferentes variáveis clínicas independentemente da autoestima dos adolescentes, de acordo com o escore da escala de Rosenberg. Resultados: Entre 1.140 adolescentes estudados, a prevalência da necessidade de tratamento ortodôntico avaliado de modo objetivo pelo IOTN-DHC ocorreu em 56,9% dos indivíduos. A avaliação subjetiva do adolescente (IOTN-AC) revelou prevalência de necessidade de tratamento de 53,3%. A análise multivariada mostrou que, além de todos os problemas dentários, o componente objetivo (IOTN-DHC), seguido do componente estético subjetivo (IOTN-AC), teve maior impacto na autoestima dos estudantes analisados. Conclusões: A insatisfação com a aparência dental é um forte preditor de baixa autoestima na adolescência.

ABSTRACT Objective: To investigate the presence of association between objective and subjective evaluation of orthodontic treatment needs in adolescents and their impact on their self-esteem. Methods: Cross-sectional study with adolescents aged 10-17 years old in Sri Ganganagar city, Rajasthan, India. The objective index of orthodontic treatment need (IOTN) dental health component (DHC) and the subjective aesthetic component (AC) were used to determine the normative and the self-perception need for orthodontic treatment, respectively. The selected students were further examined for dental trauma, tooth loss, and dental caries. Rosenberg Self-Esteem Scale was applied for self-esteem level determination. Linear regression analysis was executed to test the individual association of different independent clinical variables with self-esteem scores. Results: Among 1,140 studied adolescents, the prevalence of dental normative orthodontic treatment need was in 56.9% of individuals, whereas 53.3% of individuals considered themselves as needy for the treatment. Multivariate analyses revealed that out of all dental disorders, DHC followed by AC of IOTN had maximum impact on the self-esteem of the adolescence. Conclusions: Dissatisfaction with dental appearance is a strong predictor for low self-esteem in adolescence.

Zika Virus: Transmission, Detection, Control, and Prevention.

Zika virus (ZIKV) is a mosquito-borne Flavivirus discovered in Uganda in the 1940s. To date, three major ZIKV outbreaks have been reported. ZIKV infections have known to be primarily asymptomatic while causing mild illness in a few cases. However, the recent emergence and spread of ZIKV in the Americas has resulted in the declaration of "Public Health Emergency of International Concern" due to the potential association between the infection and prenatal microcephaly or other brain anomalies. In Brazil, a 20-fold increase in prenatal microcephaly cases and 19% increase in Guillain-Barré Syndrome (GBS) cases were reported in 2015, as compared to the preceding year. The probable deleterious effects of ZIKV infection prompt the urgent development of diagnostics and therapeutics. To this end, the existing evidences supporting the increasingly common prenatal microcephaly and GBS association and the current known ZIKV transmission dynamics, modes of detection (molecular and serology-based), and current control strategies are summarized in this review. This review also emphasizes the importance of understanding ZIKV transmission in order to design a sensitive yet cost and time-efficient detection technique. Development of an efficient detection technique would subsequently allow for better surveillance and control of ZIKV infection. Currently, limited literature is available on the pathogenesis of ZIKV, hence, focusing on the modes of ZIKV transmission could potentially contribute to the understanding of the disease spectrum and formulation of targeted treatment and control.

Inhibition of the Human ABC Efflux Transporters P-gp and BCRP by the BDE-47 Hydroxylated Metabolite 6-OH-BDE-47: Considerations for Human Exposure.

High body burdens of polybrominated diphenyl ethers (PBDEs) in infants and young children have led to increased concern over their potential impact on human development. PBDE exposure can alter the expression of genes involved in thyroid homeostasis, including those of ATP-binding cassette (ABC) transporters, which mediate cellular xenobiotic efflux. However, little information exists on how PBDEs interact with ABC transporters such as P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP). The purpose of this study was to evaluate the interactions of 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) and its hydroxylated metabolite 6-OH-BDE-47 with P-gp and BCRP, using human MDR1- and BCRP-expressing membrane vesicles and stably transfected NIH-3T3-MDR1 and MDCK-BCRP cells. In P-gp membranes, BDE-47 did not affect P-gp activity; however, 6-OH-BDE-47 inhibited P-gp activity at low µM concentrations (IC50 = 11.7 µM). In BCRP membranes, BDE-47 inhibited BCRP activity; however, 6-OH-BDE-47 was a stronger inhibitor [IC50 = 45.9 µM (BDE-47) vs. IC50 = 9.4 µM (6-OH-BDE-47)]. Intracellular concentrations of known P-gp and BCRP substrates [(3H)-paclitaxel and (3H)-prazosin, respectively] were significantly higher (indicating less efflux) in NIH-3T3-MDR1 and MDCK-BCRP cells in the presence of 6-OH-BDE-47, but not BDE-47. Collectively, our results indicate that the BDE-47 metabolite 6-OH-BDE-47 is an inhibitor of both P-gp and BCRP efflux activity. These findings suggest that some effects previously attributed to BDE-47 in biological systems may actually be due to 6-OH-BDE-47. Considerations for human exposure are discussed.

Plasma protein binding limits the blood brain barrier permeation of the pyrethroid insecticide, deltamethrin.

Previous pharmacokinetic studies of deltamethrin (DLM) have revealed that brain levels of this highly lipophilic pyrethroid insecticide are only 15-20% of plasma levels. Experiments were performed to assess determinants limiting CNS access including plasma protein binding and the efflux transporter, P-gp. A human brain microvascular endothelial cell line, hCMEC/D3, was utilized as a model in vitro system to evaluate blood-brain barrier (BBB) permeation. Incubation of DLM with a series of human serum albumin (HSA) concentrations showed that unbound (fu) DLM ranged from 80% with 0.01% HSA to ∼20% at the physiologically-relevant 4% HSA. A positive correlation (R=0.987) was seen between fu and cellular uptake. Concentration-dependent uptake of DLM in 0.01% HSA was non-linear and was reduced at 4°C and by the P-gp inhibitor cyclosporine (CSA), indicative of a specific transport process. Cellular accumulation of [(3)H]-paclitaxel, a P-glycoprotein (P-gp) substrate, was increased by CSA but not by DLM, suggesting that DLM is neither a substrate nor an inhibitor of P-gp. The concentration-dependent uptake of DLM from 4% HSA was linear and not significantly impacted by temperature or CSA. In situ brain perfusion studies monitoring brain association of DLM at 0.01% and 4% HSA confirmed the aforementioned in vitro findings. This study demonstrates that brain uptake of DLM under normal physiological conditions appears to be a passive, non-saturable process, limited by the high protein binding of the pyrethroid.

Synthesis of lipoteichoic acids in Bacillus anthracis.

Lipoteichoic acid (LTA), a glycerol phosphate polymer, is a component of the envelope of Gram-positive bacteria that has hitherto not been identified in Bacillus anthracis, the causative agent of anthrax. LTA synthesis in Staphylococcus aureus and other microbes is catalyzed by the product of the ltaS gene, a membrane protein that polymerizes polyglycerol phosphate from phosphatidyl glycerol. Here we identified four ltaS homologues, designated ltaS1 to -4, in the genome of Bacillus anthracis. Polyglycerol phosphate-specific monoclonal antibodies were used to detect LTA in the envelope of B. anthracis strain Sterne (pXO1(+) pXO2(-)) vegetative forms. B. anthracis mutants lacking ltaS1, ltaS2, ltaS3, or ltaS4 did not display defects in growth or LTA synthesis. In contrast, B. anthracis strains lacking both ltaS1 and ltaS2 were unable to synthesize LTA and exhibited reduced viability, altered envelope morphology, aberrant separation of vegetative forms, and decreased sporulation efficiency. Expression of ltaS1 or ltaS2 alone in B. anthracis as well as in other microbes was sufficient for polyglycerol phosphate synthesis. Thus, similar to S. aureus, B. anthracis employs LtaS enzymes to synthesize LTA, an envelope component that promotes bacterial growth and cell division.