Prospective Assessment of Symptoms to Evaluate Asymptomatic SARS-CoV-2 Infections in a Cohort of Health Care Workers.

BACKGROUND: The frequency of asymptomatic severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections is unclear and may be influenced by how symptoms are evaluated. In this study, we sought to determine the frequency of asymptomatic SARS-CoV-2 infections in a prospective cohort of health care workers (HCWs). METHODS: A prospective cohort of HCWs, confirmed negative for SARS-CoV-2 exposure upon enrollment, were evaluated for SARS-CoV-2 infection by monthly analysis of SARS-CoV-2 antibodies as well as referral for polymerase chain reaction testing whenever they exhibited symptoms of coronavirus disease 2019 (COVID-19). Participants completed the standardized and validated FLU-PRO Plus symptom questionnaire scoring viral respiratory disease symptom intensity and frequency at least twice monthly during baseline periods of health and each day they had any symptoms that were different from their baseline. RESULTS: Two hundred sixty-three participants were enrolled between August 25 and December 31, 2020. Through February 28, 2021, 12 participants were diagnosed with SARS-CoV-2 infection. Symptom analysis demonstrated that all 12 had at least mild symptoms of COVID-19, compared with baseline health, near or at time of infection. CONCLUSIONS: These results suggest that asymptomatic SARS-CoV-2 infection in unvaccinated, immunocompetent adults is less common than previously reported. While infectious inoculum doses and patient factors may have played a role in the clinical manifestations of SARS-CoV-2 infections in this cohort, we suspect that the high rate of symptomatic disease was due primarily to participant attentiveness to symptoms and collection of symptoms in a standardized, prospective fashion. These results have implications for studies that estimate SARS-CoV-2 infection prevalence and for public health measures to control the spread of this virus.

Durability of Antibody Response and Frequency of SARS-CoV-2 Infection 6 Months after COVID-19 Vaccination in Healthcare Workers.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies decay but persist 6 months postvaccination; lower levels of neutralizing titers persist against Delta than wild-type virus. Of 227 vaccinated healthcare workers tested, only 2 experienced outpatient symptomatic breakthrough infections, despite 59/227 exhibiting serologic evidence of SARS-CoV-2 infection, defined as presence of nucleocapsid protein antibodies.

Adverse Effects and Antibody Titers in Response to the BNT162b2 mRNA COVID-19 Vaccine in a Prospective Study of Healthcare Workers.

BACKGROUND: The relationship between postvaccination symptoms and strength of antibody responses is unclear. The goal of this study was to determine whether adverse effects caused by vaccination with the Pfizer/BioNTech BNT162b2 vaccine are associated with the magnitude of vaccine-induced antibody levels. METHODS: We conducted a single-center, observational cohort study consisting of generally healthy adult participants that were not severely immunocompromised, had no history of coronavirus disease 2019, and were seronegative for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein before vaccination. Severity of vaccine-associated symptoms was obtained through participant-completed questionnaires. Testing for immunoglobulin G antibodies against SARS-CoV-2 spike protein and receptor-binding domain was conducted using microsphere-based multiplex immunoassays performed on serum samples collected at monthly visits. Neutralizing antibody titers were determined by microneutralization assays. RESULTS: Two hundred six participants were evaluated (69.4% female, median age 41.5 years old). We found no correlation between vaccine-associated symptom severity scores and vaccine-induced antibody titers 1 month after vaccination. We also observed that (1) postvaccination symptoms were inversely correlated with age and weight and more common in women, (2) systemic symptoms were more frequent after the second vaccination, (3) high symptom scores after first vaccination were predictive of high symptom scores after second vaccination, and (4) older age was associated with lower titers. CONCLUSIONS: Lack of postvaccination symptoms after receipt of the BNT162b2 vaccine does not equate to lack of vaccine-induced antibodies 1 month after vaccination.

A three-antigen Plasmodium falciparum DNA prime-Adenovirus boost malaria vaccine regimen is superior to a two-antigen regimen and protects against controlled human malaria infection in healthy malaria-naïve adults.

BACKGROUND: A DNA-prime/human adenovirus serotype 5 (HuAd5) boost vaccine encoding Plasmodium falciparum (Pf) circumsporozoite protein (PfCSP) and Pf apical membrane antigen-1 (PfAMA1), elicited protection in 4/15 (27%) of subjects against controlled human malaria infection (CHMI) that was statistically associated with CD8+ T cell responses. Subjects with high level pre-existing immunity to HuAd5 were not protected, suggesting an adverse effect on vaccine efficacy (VE). We replaced HuAd5 with chimpanzee adenovirus 63 (ChAd63), and repeated the study, assessing both the two-antigen (CSP, AMA1 = CA) vaccine, and a novel three-antigen (CSP, AMA1, ME-TRAP = CAT) vaccine that included a third pre-erythrocytic stage antigen [malaria multiple epitopes (ME) fused to the Pf thrombospondin-related adhesive protein (TRAP)] to potentially enhance protection. METHODOLOGY: This was an open label, randomized Phase 1 trial, assessing safety, tolerability, and VE against CHMI in healthy, malaria naïve adults. Forty subjects (20 each group) were to receive three monthly CA or CAT DNA priming immunizations, followed by corresponding ChAd63 boost four months later. Four weeks after the boost, immunized subjects and 12 infectivity controls underwent CHMI by mosquito bite using the Pf3D7 strain. VE was assessed by determining the differences in time to parasitemia as detected by thick blood smears up to 28-days post CHMI and utilizing the log rank test, and by calculating the risk ratio of each treatment group and subtracting from 1, with significance calculated by the Cochran-Mantel-Haenszel method. RESULTS: In both groups, systemic adverse events (AEs) were significantly higher after the ChAd63 boost than DNA immunizations. Eleven of 12 infectivity controls developed parasitemia (mean 11.7 days). In the CA group, 15 of 16 (93.8%) immunized subjects developed parasitemia (mean 12.0 days). In the CAT group, 11 of 16 (63.8%) immunized subjects developed parasitemia (mean 13.0 days), indicating significant protection by log rank test compared to infectivity controls (p = 0.0406) and the CA group (p = 0.0229). VE (1 minus the risk ratio) in the CAT group was 25% compared to -2% in the CA group. The CA and CAT vaccines induced robust humoral (ELISA antibodies against CSP, AMA1 and TRAP, and IFA responses against sporozoites and Pf3D7 blood stages), and cellular responses (IFN-γ FluoroSpot responses to CSP, AMA1 and TRAP) that were not associated with protection. CONCLUSIONS: This study demonstrated that the ChAd63 CAT vaccine exhibited significant protective efficacy, and confirmed protection was afforded by adding a third antigen (T) to a two-antigen (CA) formulation to achieve increased VE. Although the ChAd63-CAT vaccine was associated with increased frequencies of systemic AEs compared to the CA vaccine and, historically, compared to the HuAd5 vectored malaria vaccine encoding CSP and AMA1, they were transient and associated with increased vector dosing.

Adverse effects and antibody titers in response to the BNT162b2 mRNA COVID-19 vaccine in a prospective study of healthcare workers.

BACKGROUND: mRNA COVID-19 vaccines are playing a key role in controlling the COVID-19 pandemic. The relationship between post-vaccination symptoms and strength of antibody responses is unclear. OBJECTIVE: To determine whether adverse effects caused by vaccination with the Pfizer/BioNTech BNT162b2 vaccine are associated with the magnitude of vaccine-induced antibody levels. DESIGN: Single center, prospective, observational cohort study. SETTING: Participants worked at Walter Reed National Military Medical Center and were seen monthly at the Naval Medical Research Center Clinical Trials Center. PARTICIPANTS: Generally healthy adults that were not severely immunocompromised, had no history of COVID-19, and were seronegative for SARS-CoV-2 spike protein prior to vaccination. MEASURES: Severity of vaccine-associated symptoms was obtained through participant completed questionnaires. Testing for IgG antibodies against SARS-CoV-2 spike protein and receptor binding domain was conducted using microsphere-based multiplex immunoassays. RESULTS: 206 participants were evaluated (69.4% female, median age 41.5 years old). We found no correlation between vaccine-associated symptom severity scores and vaccine-induced antibody titers one month after vaccination. We also observed that 1) post-vaccination symptoms were inversely correlated with age and weight and more common in women, 2) systemic symptoms were more frequent after the second vaccination, 3) high symptom scores after first vaccination were predictive of high symptom scores after second vaccination, and 4) older age was associated with lower titers. LIMITATIONS: Study only observes antibody responses and consists of healthy participants. CONCLUSIONS: Lack of post-vaccination symptoms following receipt of the BNT162b2 vaccine does not equate to lack of vaccine-induced antibodies one month after vaccination. This study also suggests that it may be possible to design future mRNA vaccines that confer robust antibody responses with lower frequencies of vaccine-associated symptoms. FUNDING: This study was executed by the Infectious Disease Clinical Research Program (IDCRP), a Department of Defense (DoD) program executed by the Uniformed Services University of the Health Sciences (USUHS) through a cooperative agreement by the Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc. (HJF). This project has been funded by the Defense Health Program, U.S. DoD, under award HU00012120067. Project funding for JHP was in whole or in part with federal funds from the National Cancer Institute, National Institutes of Health, under Contract No. HHSN261200800001E. The funding bodies have had no role in the study design or the decision to submit the manuscript for publication.

Prospective Assessment of SARS-CoV-2 Seroconversion (PASS) study: an observational cohort study of SARS-CoV-2 infection and vaccination in healthcare workers.

BACKGROUND: SARS-CoV-2 is a recently emerged pandemic coronavirus (CoV) capable of causing severe respiratory illness. However, a significant number of infected people present as asymptomatic or pauci-symptomatic. In this prospective assessment of at-risk healthcare workers (HCWs) we seek to determine whether pre-existing antibody or T cell responses to previous seasonal human coronavirus (HCoV) infections affect immunological or clinical responses to SARS-CoV-2 infection or vaccination. METHODS: A cohort of 300 healthcare workers, confirmed negative for SARS-CoV-2 exposure upon study entry, will be followed for up to 1 year with monthly serology analysis of IgM and IgG antibodies against the spike proteins of SARS-CoV-2 and the four major seasonal human coronavirus - HCoV-OC43, HCoV-HKU1, HCoV-229E, and HCoV-NL63. Participants will complete monthly questionnaires that ask about Coronavirus Disease 2019 (COVID-19) exposure risks, and a standardized, validated symptom questionnaire (scoring viral respiratory disease symptoms, intensity and severity) at least twice monthly and any day when any symptoms manifest. SARS-CoV-2 PCR testing will be performed any time participants develop symptoms consistent with COVID-19. For those individuals that seroconvert and/or test positive by SARS-CoV-2 PCR, or receive the SARS-CoV-2 vaccine, additional studies of T cell activation and cytokine production in response to SARS-CoV-2 peptide pools and analysis of Natural Killer cell numbers and function will be conducted on that participant's cryopreserved baseline peripheral blood mononuclear cells (PBMCs). Following the first year of this study we will further analyze those participants having tested positive for COVID-19, and/or having received an authorized/licensed SARS-CoV-2 vaccine, quarterly (year 2) and semi-annually (years 3 and 4) to investigate immune response longevity. DISCUSSION: This study will determine the frequency of asymptomatic and pauci-symptomatic SARS-CoV-2 infection in a cohort of at-risk healthcare workers. Baseline and longitudinal assays will determine the frequency and magnitude of anti-spike glycoprotein antibodies to the seasonal HCoV-OC43, HCoV-HKU1, HCoV-229E, and HCoV-NL63, and may inform whether pre-existing antibodies to these human coronaviruses are associated with altered COVID-19 disease course. Finally, this study will evaluate whether pre-existing immune responses to seasonal HCoVs affect the magnitude and duration of antibody and T cell responses to SARS-CoV-2 vaccination, adjusting for demographic covariates.

Refinement of the CS6-expressing enterotoxigenic Escherichia coli strain B7A human challenge model: A randomized trial.

BACKGROUND: Human challenge models for enterotoxigenic Escherichia coli (ETEC) facilitate vaccine down-selection. The B7A (O148:H28 CS6+LT+ST+) strain is important for vaccine development. We sought to refine the B7A model by identifying a dose and fasting regimen consistently inducing moderate-severe diarrhea. METHODS: An initial cohort of 28 subjects was randomized (1:1:1:1) to receive B7A following an overnight fast at doses of 108 or 109 colony forming units (cfu) or a 90-minute fast at doses of 109 or 1010 cfu. A second cohort included naïve and rechallenged subjects who had moderate-severe diarrhea and were given the target regimen. Immune responses to important ETEC antigens were assessed. RESULTS: Among subjects receiving 108 cfu of B7A, overnight fast, or 109 cfu, 90-minute fast, 42.9% (3/7) had moderate-severe diarrhea. Higher attack rates (71.4%; 5/7) occurred in subjects receiving 109 cfu, overnight fast, or 1010 cfu, 90-minute fast. Upon rechallenge with 109 cfu of B7A, overnight fast, 5/11 (45.5%) had moderate-severe diarrhea; the attack rate among concurrently challenge naïve subjects was 57.9% (11/19). Anti-CS6, O148 LPS and LT responses were modest across all groups. CONCLUSIONS: An overnight fast enabled a reduction in the B7A inoculum dose; however, the attack rate was inconsistent and protection upon rechallenge was minimal.

Antimicrobial resistance (AMR) and molecular characterization of Neisseria gonorrhoeae in Ghana, 2012-2015.

Neisseria gonorrhoeae antimicrobial resistance (AMR) surveillance is essential for tracking the emergence and spread of AMR strains in local, national and international populations. This is crucial for developing or refining treatment guidelines. N. gonorrhoeae multiantigen sequence typing (NG-MAST) is beneficial for describing the molecular epidemiology of gonococci at national and international levels. Elucidation of AMR determinants to ß-lactam drugs, is a means of monitoring the development of resistance. In Ghana, little is known about the current gonococcal AMR prevalence and no characterization of gonococcal isolates has been previously performed. In this study, gonococcal isolates (n = 44) collected from five health facilities in Ghana from 2012 to 2015, were examined using AMR testing, NG-MAST and sequencing of penA. High rates of resistance were identified to tetracycline (100%), benzylpenicillin (90.9%), and ciprofloxacin (81.8%). One isolate had a high cefixime MIC (0.75 µg/ml). Twenty-eight NG-MAST sequence types (STs) were identified, seventeen of which were novel. The isolate with the high cefixime MIC contained a mosaic penA-34 allele and belonged to NG-MAST ST1407, an internationally spreading multidrug-resistant clone that has accounted for most cefixime resistance in many countries. In conclusion, AMR testing, NG-MAST, and sequencing of the AMR determinant penA, revealed high rates of resistance to tetracycline, benzylpenicillin, and ciprofloxacin; as well as a highly diverse population of N. gonorrhoeae in Ghana. It is imperative to continue with enhanced AMR surveillance and to understand the molecular epidemiology of gonococcal strains circulating in Ghana and other African countries.

Review: chronic and persistent diarrhea with a focus in the returning traveler.

BACKGROUND: Travelers' diarrhea is a common malady afflicting up to 50% of travelers after a 2-week travel period. An appreciable percentage of these cases will become persistent or chronic. We summarized the published literature reporting persistent/chronic diarrhea in travelers elucidating current understanding of disease incidence, etiology and regional variability. METHODS: We searched electronic databases (Medline, Embase, and Cochrane database of clinical trials) from 1990 to 2015 using the following terms: "chronic or persistent diarrh* and (returning) travel* or enteropathogen, GeoSentinel, and travel-associated infection. Included studies published in the English language on adult returning travelers (duration < 3-months) reporting denominator data. Point estimates and standard 95% confidence intervals were calculated for incidence using a random-effects model. Study incidence heterogeneity rates were assessed using x2 heterogeneity statistics, graphically represented with Forest plots. RESULTS: We identified 19 studies meeting the inclusion criteria (all published after 1999). 18 studies reported upon the incidence of persistent/chronic diarrhea as a syndromic diagnosis in returning travelers; one study reported adequate denominator data from which to assess pathogen specific etiology. Giardiasis comprise an appreicaible percentage of infectious mediated persistent/chronic diarrhea in returning travelers. The overall estimate of persistent/chronic diarrhea incidence was 6% (0.05-0.07) in 321,454, travelers; with significant heterogeniety observed across regions. The total number of regional travelers, and point estimates for incidence (95% CI) for Latin American, African, and Asian travelers were [15816 (0.09 [0.07-0.11]), 42290 (0.06 [0.05-0.07]), and 27433 (0.07 [0.06-0.09])] respectively. We identified lower published rates of chronic diarrhea from Sub-Saharan Africa relative to [North Africa, South Central Asia, and Central America]. Persistent/chronic diarrhea ranked fourth as a syndromic diagnosis in all regions. CONCLUSIONS: Persistent/Chronic diarrhea is a leading syndromic diagnosis in returning travelers across all regions. The 6% incidence [proportionate morbidity (PM) of 60] observed in over >300,000 global travelers is comparable to prior estimates. We identified lower published rates of chronic diarrhea from Sub-Saharan Africa relative to [North Africa, South Central Asia, and Central America]. Giardiasis comprises an appreciabile percentatge of travel-associated infectious mediated persistent/chronic diarrhea. There's a dearth of published data characterizing the incidence of specific enteropathogenic etiologies for persistent/chronic diarrhea in returning travelers.

Melioidosis in lower provincial Cambodia: A case series from a prospective study of sepsis in Takeo Province.

Melioidosis is a severe infectious disease caused by the gram-negative soil bacterium Burkholderia pseudomallei. Melioidosis is well known to be a major cause of morbidity and mortality in Southeast Asia, particularly in Thailand. However, melioidosis remains underreported in surrounding areas such as Cambodia. We report a case series of melioidosis in seven patients from Takeo Province, Cambodia. The patients, aged 24-65 years, were enrolled from May 2014 to May 2015 during a one year prospective study of sepsis at Takeo Provincial Hospital. They presented with fever, rigors, dyspnea, fatigue, diaphoresis, productive cough, and skin abscesses. Six of the seven patients were also hyponatremic. B. pseudomallei was cultured from the blood of six patients and the sputum of one patient. In this manuscript, we provide a detailed description of the clinical presentation, case management and laboratory confirmation of B. pseudomallei, as well as discuss the difficulties of identifying and treating melioidosis in low resource settings.

Clostridium sphenoides Chronic Osteomyelitis Diagnosed Via Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry, Conflicting With 16S rRNA Sequencing but Confirmed by Whole Genome Sequencing.

We report a case of successful treatment of chronic osteomyelitis (emanating from contaminated soil exposure) caused by Clostridium sphenoides, an organism infrequently identified as a cause of human infection and more saliently osteomyelitis (only 1 reported case in the literature). Additional impetus for reporting this case resides in the insights gained regarding pathogen identification exploiting sophisticated molecular platforms coupled to traditional microbial culture-based methods. The fastidious nature of cultivating anaerobic organisms required initial attempts at 16S rRNA sequencing to identify a Clostridium species (Clostridium celerecrescens). However, on exploiting matrix-assisted laser desorption ionization time of flight (MALDI TOF) technology, C. sphenoides was identified, and confirmed on whole genome sequencing. The discrepancies noted in the varying platforms require vigilance to seek complementary testing for conflicting results. Although highly accurate, the MALDI TOF and 16S rRNA sequencing platforms are not immune to false identification particularly in differentiating closely related organisms. More germane, whole genome sequencing should be entertained when conflicting results are obtained from MALDI TOF and 16S rRNA sequencing. Precise species and/or strain level identification can be clinically relevant as antimicrobial sensitivity profiles may be discrepant between closely related species influencing clinical outcomes. Thus, it is incumbent on us to strive to acquire the correct species characterization when resources allow to dictate optimal treatment.

Rectal cultures before transrectal ultrasound-guided prostate biopsy reduce post-prostatic biopsy infection rates.

OBJECTIVE: To test our hypothesis that a targeted rectal screening protocol before transrectal ultrasound (TRUS)-guided biopsy would potentiate streamlined prophylaxis, thereby reducing postbiopsy infectious rates while minimizing unnecessary broad-spectrum antibiotic use. To this end, we instituted preprocedure rectal cultures in an effort to identify fluoroquinolone (FQ)- resistant flora using selective media to optimally direct targeted prophylactic antibiotic administration. The inexorably increasing prevalence of multidrug-resistant microorganisms, notably extended spectrum beta lactamase (ESBL)-producing and FQ-resistant Enterobacteriaceae has increased the post-TRUS prostatic biopsy infection rates, including life-threatening sepsis. METHODS: A total of 235 rectal swabs were obtained and plated directly onto MacConkey agar plates containing 10-µg/mL ciprofloxacin. Following the screening procedure, antimicrobial susceptibility results were used to develop a customized antibiotic prophylaxis regimen to be administered before biopsy. Following the biopsy procedure, the patients were seen in follow-up within 7 days, and information was gathered on potential adverse effects, clinical appointments for infections, and potential antibiotics received. RESULTS: Thirty-two-patients (14%) had FQ-resistant isolates (most Escherichia coli), and 3 (1.3%) were ESBL-producing isolates. There were no infectious complications identified in this period, (compared with 3 septic complications among 103 biopsies in the 4 months preceding the study). CONCLUSION: Rectal cultures obtained before TRUS biopsy, using selective media to identify FQ-resistant Enterobacteriaceae, facilitate targeted antibiotic prophylaxis, and appear to be highly efficacious in reducing infectious complications.

Two cases illustrating successful adjunctive interferon-γ immunotherapy in refractory disseminated coccidioidomycosis.

Protective immunity and host resistance to coccidioidomycosis require a robust cell-mediated immunity with adequate production of Th1 cytokines including interleukin-12, and IFN-γ and appropriate regulation and coordinated functionality of Th1/Th2 responses and IL-12/IFN-γ cytokine axes. IFN-γ augments the anti-fungal activity of effector immune cells against a variety of fungi. Numerous animal models have demonstrated the potential efficacy of adjunctive IFN-γ in treatment of invasive mycoses. Yet, despite these promising data, a paucity of literature documents efficacious adjunctive IFN-γ administration in refractory coccidioidomycosis. We present two cases of refractory disease occurring at our institution who responded to adjunctive IFN-γ.

Inhalational diesel exhaust exposure in submariners: observational study.

OBJECTIVE: An observational study was performed with a convenience sample of 38 submariners exposed to diesel exhaust for 9 hours, to assess the development of reactive airways dysfunction syndrome (RADS) after prophylactic corticosteroid treatment. METHODS: Twenty-four subjects were available for baseline physical examinations, pulmonary function tests, and chest radiographs, and 16 more subjects were available for interviews; 30 subjects were available for 6-month follow-up surveys. Subjects were treated on the basis of presenting symptoms; 19 subjects were treated with a 10-day course of orally administered prednisone, accompanied by 30 days of inhaled fluticasone/salmeterol therapy. RESULTS: There were no cases of RADS diagnosed at 6-month follow-up evaluations. CONCLUSION: There were no cases of RADS diagnosed at 6-month follow-up evaluations in submariners with uncontrolled, isolated, heavy diesel exhaust exposure, despite many initial symptoms that portended the diagnosis. To our knowledge, this is the largest reported case study of corticosteroid treatment initiated with an expressed intention to prevent the development of RADS after an isolated diesel exhaust exposure. Although we cannot prove that early intervention with corticosteroids prevented RADS, we think that the implementation of prompt prophylactic treatment expedited symptom resolution and might have prevented RADS development, on the basis of previous historical control data. RADS resulting from diesel exhaust may be an important public health issue, and our hope is to promote increased recognition of the diagnosis, which often is not suspected upon initial presentation but is delayed by up to several years. Increasing awareness may prompt pursuit of more-aggressive interventions with acute and protracted corticosteroid treatment and execution of the necessary controlled trials to establish treatment efficacy in mitigating the severity and/or circumventing the development of RADS.

Investigating the potential of statin medications as a nitric oxide (NO) release agent to decrease decompression sickness: a review article.

Understanding the biochemical mechanisms influencing bubble pathophysiology may foster novel pharmacologic non-recompressive strategies that may prevent, ameliorate, and treat decompression sickness (DCS), and the injury sustained from arterial gas emboli (AGE) encountered in hyperbaric and hypobaric exposures, as well as in surgery and trauma. This review explores the biochemical effects of nitric oxide (NO) release agents, their potential impact on bubble pathophysiology, and possible use as a pharmacological intervention to reduce DCS risk and AGE injury. The hypotheses discussed contend that exogenous NO administration or mediators of endogenous NO up-regulation may reduce DCS risk and severity by mediating; (1) decreased populations of gaseous nuclei, (2) decreased bubble nuclei adherence, (3) depression of the deleterious bubble-mediated inflammatory and coagulation cascades and (4) preservation of endothelial integrity, which may defend against bubble-mediated injury. Statin medications alter numerous biochemical, and biophysical processes, which may influence bubble formation. Statins preserve endothelial integrity, reduce ischemia/reperfusion injury, and depress the interdependent inflammatory and coagulation cascades via pleiotropic properties involving up-regulation of endothelial nitric oxide synthase (eNOS) and NO. Numerous studies are researching statins, for their potential efficacy in reducing primary and secondary morbidity and mortality from cardiocerebrovascular, inflammatory (autoimmune), and infectious (sepsis) disease. Additionally, statin-mediated lipid reduction may reduce bubble generation via alterations in plasma "rheology", and surface tension. The statins are attractive potential NO release with minimal adverse side effects, and proven long-term safety, that may potentially mitigate the risk and severity of DCS. We will elaborate on the insight gained into the mechanisms proven and hypothesized for NO-mediated reductions in bubble formation, and DCS incidence and severity, with a focus on the potential for statin medications, in addition to the direct NO-donor medications such as isosorbide mononitrate and nitroglycerine.

Prophylactic statins as a possible method to decrease bubble formation in diving.

INTRODUCTION: Nitric oxide (NO) may decrease bubble formation in diving. Statin medications are attractive potential options to increase NO. Statins exhibit a proven safety profile, and possess a myriad of pleiotropic properties improving vascular endothelial function. Additionally, statin-mediated lipid reduction may reduce bubble generation via alterations in plasma surface tension. We investigated the efficacy of atorvastatin as a pharmacological intervention to reduce the risk of bubble formation after diving, a surrogate for decompression sickness (DCS). METHODS: There were 16 trained military divers who completed a provacative hyperbaric chamber dive protocol after taking either 80-mg of atorvastatin or placebo for 4 d. Subjects completed the alternate medication regimen no sooner than 2 wk. After each dive, subjects were subjected to precordial trans-thoracic echocardiographic exams via standardized protocols. Bubbles were graded via a non-parametric, ordinal grading system and statistically analyzed via Wilcoxon signed-rank tests. RESULTS: We found no within subject differences for the maximum bubble grade scores (z = 0.00, p = 1.00, n=16). Low-density lipoprotein (LDL), and total cholesterol (TC) levels decreased significantly (107.6 +/- 26.2 to 79.3 +/- 21.9 mg x dl(-1) and 175 +/- 20.9 to 147 +/- 22.4 mg x dl(-1), respectively) 1-2 wk post statin administration. Age, bioelectrical impedance (BEI), TC, LDL, potassium, and calcium demonstrated positive correlations to placebo bubble grades. DISCUSSION: Prophylactic 80-mg atorvastatin administration for 4 d failed to reduce the number of intravascular bubbles observed following a 60-ft, 80-min dry chamber dive despite significant acute reductions in lipid levels. Several hypotheses may explain why statins failed to decrease bubble volume: (1) differential influence of statins on the venous vs. arterial vasculature; (2) failure to elicit an improvement in endothelial function and, therefore, the hypothesized endothelial conditioning in younger patients possessing normal baseline; and (3) the ordinal grading system encompassing a substantial variation in bubble volume (bubbles Scm(-2)).

Vitamin D supplementation in underway submariners.

INTRODUCTION: We assessed the efficacy of vitamin D supplementation in maintaining normal calcium and bone homeostasis in underway submariners deprived of sunlight. METHODS: Serum 25-hydroxycholecalciferol (25(OH)D), 1,25-dihydroxycholecalciferol (1,25(OH)2D), calcium, parathyroid hormone (PTH), phosphate, osteocalcin, bone specific alkaline phosphatase, and urinary levels of N-telopeptide were examined in 51 subjects aboard a submarine. These levels were obtained prior to a 76-d deployment, before and after a 6-d liberty period (deployment day 49 and 55), and on return to homeport. There were 26 subjects who received 400-lU of vitamin D daily supplementation, and 25 who received placebo. RESULTS: Both groups exhibited significant reductions in 25(OH)D levels in the initial submergence (a decrease from 28.3 +/- 15 ng x ml(-1) to 24.1 +/- 10 ng x ml(-1) in the experimental group and 26.3 +/- 10 ng x ml(-1) to 20.7 +/- 9 ng x ml(-1) in the controls), an increase in 25(OH)D levels not significantly different from baseline during the liberty period, and decrements in 25(OH)D on repeat submergence (22.8 +/- 10 ng x ml(-1) in experimental and 21.4 +/- 10 ng x ml(-1) in controls). Both groups exhibited an increase in post-liberty osteocalcin (20.4 +/- 6 ng x ml(-1) to 24.5 +/- 5 ng x ml(-1) for experimental and 18.3 +/- 6 to 23.5 +/- 7 ng x ml(-1) for controls), and stable serum calcium levels throughout the patrol. CONCLUSIONS: 400-IU daily vitamin D supplementation was insufficient in maintaining serum vitamin D levels in underway submariners, engendering biochemical evidence of bone resorption and turnover. Six d sunlight exposure compensated for 49 d sunlight absence, supporting the enormous capacity of UV-B mediated vitamin D production.