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Madopar, a combination of levodopa with benserazide, induced an inconsistent rise in plasma growth hormone in unmedicated patients with Parkinson's disease and in controls, and a greater growth hormone rise in Parkinsonian subjects on chronic Madopar therapy. In subjects on chronic therapy with levodopa and carbidopa (Sinemet), the growth hormone releasing effect of Madopar was blunted. Madopar increased plasma prolactin (PRL) in controls, unmedicated patients and patients on Madopar therapy while in patients on Sinemet therapy the PRL-releasing effect of Madopar was strikingly reduced. Since these data were interpreted as due to a defective dopamine tone in the hypothalamus of Parkinsonian subjects on Madopar but not Sinemet therapy, a direct dopamine receptor agonist, lisuride was administered. Lisuride, however, elicited a blunted growth hormone response both in patients on Madopar and Sinemet therapy, without revealing a state of supersensitivity of dopamine receptors for growth hormone control in Parkinsonian subjects on Madopar therapy. No difference was present in the PRL-lowering effect of lisuride in the different experimental groups. These findings suggest that: (1) hypothalamic dopamine function is impaired in Parkinsonian subjects on Madopar therapy, preserved in unmedicated patients and enhanced in patients on Sinemet therapy; (2) the endocrine effects observed in Parkinsonian subjects on chronic Madopar therapy may be due to some penetration of benserazide across the blood brain barrier in the region of the hypothalamus; (3) since Madopar and Sinemet are in essence equally effective antiparkinsonian remedies, penetration of benserazide does not occur across the blood brain barrier surrounding the nigrostriatal system. [ABSTRACT FROM AUTHOR]

Background: In South Africa, extragenital etiological sexually transmitted infection (STI) screening among men who have sex with men (MSM) is not routinely available. We aimed to determine the prevalence of STI pathogens at rectal and pharyngeal sites, syphilis seroprevalence, and associated risk factors among a selection of high-risk MSM without symptomatic urethritis attending a men's health clinic in Johannesburg, South Africa., Methods: A cross-sectional study was conducted in 2022. Enrolled clients self-reported demographic, sexual behavioral risks, and clinical information. Client or clinician-collected rectal and pharyngeal swabs were tested for Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, and Trichomonas vaginalis. C. trachomatis-positive rectal samples were reflex tested for lymphogranuloma venereum. Blood specimens were screened for syphilis. Univariate and multivariate regression models were used to determine factors independently associated with the presence of an extragenital STI or syphilis., Results: Among the 97 participants (median age, 29 years), 24.7% had an extragenital STI and 9.4% had high nontreponemal antibody titers (rapid plasma reagin ≥1:16). Rectal STIs were detected in 26.4% participants: N. gonorrhoeae (14.3%), C. trachomatis (9.9%), and M. genitalium (5.5%). Pharyngeal STIs were less prevalent (4.1%). Overall, the prevalence of any STI was 41%. Sex under the influence of drugs (adjusted odds ratio, 4.94; 95% confidence interval, 1.56-15.69) and engaging in condomless receptive anal intercourse with a casual partner (adjusted odds ratio, 8.36; 95% confidence interval, 1.73-40.28) were independent risk factors for having an extragenital STI., Conclusions: The high burden of extragenital STIs and active syphilis in asymptomatic MSM underscores the importance of routine etiological screening in this key population, as the syndromic approach would not enable detection or treatment of these infections., Competing Interests: Conflict of Interest and Sources of Funding: The authors have no conflicts of interest to declare. This work was internally funded by the Centre of HIV & STIs at the National Institute for Communicable Diseases, a division of the National Health Laboratory Service, South Africa., (Copyright © 2024 American Sexually Transmitted Diseases Association. All rights reserved.)

Background: In South Africa, Neisseria gonorrhoeae , which is the predominant cause of male urethritis, is treated syndromically using dual ceftriaxone and azithromycin therapy. We determined antimicrobial susceptibilities of N. gonorrhoeae isolates from urethral discharge specimens, and genetically characterised those with elevated minimum inhibitory concentrations (MICs) for first-line antimicrobials., Methods: Routine antimicrobial susceptibility testing (AST) of N. gonorrhoeae isolates included E-test for ceftriaxone, cefixime and gentamicin and agar dilution for azithromycin and spectinomycin. Neisseria gonorrhoeae Sequence Typing for Antimicrobial Resistance (NG-STAR) was performed for isolates with elevated MICs to identify antimicrobial resistance (AMR) determinants, and Neisseria gonorrhoeae Multi-Antigen Sequence Typing (NG-MAST) was used to determine strain relatedness., Results: N. gonorrhoeae was cultured from urethral discharge swab specimens obtained from 196 of 238 (82.4%) men presenting to a primary healthcare facility in Johannesburg in 2021. All viable isolates were susceptible to extended-spectrum cephalosporins. Four isolates had high azithromycin MICs ranging from 32mg/L to >256mg/L and grouped into two novel NG-MAST and NG-STAR groups. Two isolates from Group 1 (NG-MAST ST20366, NG-STAR ST4322) contained mutated mtrR (G45D) and 23S rRNA (A2059G) alleles, while the two isolates from Group 2 (NG-MAST ST20367, NG-STAR ST4323) had different mutations in mtrR (A39T) and 23S rRNA (C2611T)., Conclusions: We report the first cases of high-level azithromycin resistance in N. gonorrhoeae from South Africa. Continued AMR surveillance is critical to detect increasing azithromycin resistance prevalence in N. gonorrhoeae , which may justify future modifications to the STI syndromic management guidelines.

The prevalence of Mycoplasma genitalium (MG) and MG antimicrobial resistance (AMR) appear to be high internationally, however, prevalence data remain lacking globally. We evaluated the prevalence of MG and MG AMR-associated mutations in men who have sex with men (MSM) in Malta and Peru and women at-risk for sexually transmitted infections in Guatemala, South Africa, and Morocco; five countries in four WHO regions mostly lacking MG prevalence and AMR data, and estimated MG coinfections with Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Trichomonas vaginalis (TV). Male urine and anorectal samples, and vaginal samples were tested for MG, CT, NG, and TV (only vaginal samples) using Aptima assays (Hologic). AMR-associated mutations in the MG 23S rRNA gene and parC gene were identified using ResistancePlus MG kit (SpeeDx) or Sanger sequencing. In total, 1,425 MSM and 1,398 women at-risk were recruited. MG was detected in 14.7% of MSM (10.0% in Malta and 20.0% Peru) and in 19.1% of women at-risk (12.4% in Guatemala, 16.0% Morocco, 22.1% South Africa). The prevalence of 23S rRNA and parC mutations among MSM was 68.1 and 29.0% (Malta), and 65.9 and 5.6% (Peru), respectively. Among women at-risk, 23S rRNA and parC mutations were revealed in 4.8 and 0% (Guatemala), 11.6 and 6.7% (Morocco), and 2.4 and 3.7% (South Africa), respectively. CT was the most frequent single coinfection with MG (in 2.6% of MSM and 4.5% of women at-risk), compared to NG + MG found in 1.3 and 1.0%, respectively, and TV + MG detected in 2.8% of women at-risk. In conclusion, MG is prevalent worldwide and enhanced aetiological MG diagnosis, linked to clinical routine detection of 23S rRNA mutations, in symptomatic patients should be implemented, where feasible. Surveillance of MG AMR and treatment outcome would be exceedingly valuable, nationally and internationally. High levels of AMR in MSM support avoiding screening for and treatment of MG in asymptomatic MSM and general population. Ultimately, novel therapeutic antimicrobials and/or strategies, such as resistance-guided sequential therapy, and ideally an effective MG vaccine are essential., Competing Interests: Hologic provided the Aptima M. genitalium, Aptima Combo 2, and Aptima T. vaginalis tests (special thanks to Philip Mueller and Damon Getman). However, no financial support for the testing was obtained, and Hologic had no role in data collection, analysis, interpretation or writing the paper., (Copyright © 2023 Shipitsyna, Kularatne, Golparian, Müller, Vargas, Hadad, Padovese, Hancali, Alvarez, Oumzil, Camey, Blondeel, Toskin and Unemo.)

Background: Antimicrobial resistance in Mycoplasma genitalium is a global concern, as therapeutic options are limited. We aimed to determine the prevalence of macrolide and fluoroquinolone resistance-associated genetic determinants and strain diversity in M. genitalium-positive surveillance specimens from symptomatic primary health care center attendees in South Africa (2015-2018). A secondary objective was to investigate for an association between M. genitalium strain type, HIV serostatus, and antimicrobial resistance., Methods: A total of 196 M. genitalium-positive specimens from adult males and females presenting with genital discharge to primary health care centers were tested for resistance-associated mutations in 23S rRNA, parC and gyrA. A dual-locus sequence type (DLST) was assigned to M. genitalium strains based on the detection of single nucleotide polymorphisms in the semiconserved 5' region of the mgpB gene (MG191-sequence typing) as well as the enumeration of short tandem repeats within the lipoprotein gene (MG309 short tandem repeat typing)., Results: The A2059G mutation in 23S rRNA, associated with macrolide resistance, was detected in 3 of 182 specimens (1.7%; 95% confidence interval, 0.3-4.7). We did not detect gyrA or parC mutations associated with fluoroquinolone resistance in specimens that could be sequenced. Molecular typing with DLST revealed genetic heterogeneity, with DLST 4-11 being the most common M. genitalium strain type detected. There were no associations between DLST and macrolide resistance or HIV infection., Conclusions: We found a low prevalence of M. genitalium strains with macrolide resistance-associated mutations over a 4-year surveillance period. Ongoing antimicrobial resistance surveillance is essential for informing genital discharge syndromic treatment guidelines., Competing Interests: Conflict of Interest and Sources of Funding: None declared., (Copyright © 2022 American Sexually Transmitted Diseases Association. All rights reserved.)

Treponema pallidum macrolide resistance and clinical treatment failure have emerged rapidly within communities where macrolides have been used as convenient, oral therapeutic alternatives to benzathine penicillin G for syphilis or for other clinical indications. Macrolides are not included in the South African syndromic management guidelines for genital ulcer disease; however, in 2015, a 1-g dose of azithromycin was incorporated into treatment algorithms for genital discharge. We determined the prevalence of 23S rRNA macrolide resistance-associated point mutations in 135 T. pallidum-positive surveillance specimens from Botswana, Zimbabwe, and South Africa between 2008 and 2018. Additionally, we investigated the association between macrolide resistance, T. pallidum strain type, and HIV coinfection. A significant increase in the prevalence of the A2058G macrolide resistance-associated point mutation was observed in specimens collected after 2015. There was a high level of molecular heterogeneity among T. pallidum strains circulating in the study communities, with strain type 14d/f being the most predominant in South Africa. Fourteen novel strain types, derived from three new tpr gene restriction fragment length polymorphism patterns and seven new tp0548 gene sequence types, were identified. There was an association between A2058G-associated macrolide resistance and T. pallidum strain types 14d/f and 14d/g but no association between T. pallidum macrolide resistance and HIV coinfection. The majority of T. pallidum strains, as well as strains containing the A2058G mutation, belonged to the SS14-like clade. This is the first study to extensively detail the molecular epidemiology and emergence of macrolide resistance in T. pallidum in southern Africa.

Objectives: Micro ribonucleic acids (miRNAs) are small non-coding RNA molecules that control gene expression by translational inhibition. Exercise has been shown to affect several miRNAs' expression in healthy subjects, but this has not yet been studied in patients with coronary artery disease (CAD). Since exercise training confers beneficial long-term effects and may also trigger acute coronary events, it is of utmost interest to be able to identify those who are risk for untoward effects. Therefore, we set out to assess miRNA expression in response to maximal ergospirometry in patients with CAD., Methods: Total RNA was extracted from blood drawn immediately before and 5 min after maximal cycle-ergospirometry (10 male and 10 female CAD patients). A qRT-PCR was performed for 187 target miRNAs associated with endothelial function/dysfunction, cardiovascular disease, myocardial infarction, and sudden cardiac death., Results: In response to a maximal ergospirometry, 33 miRNAs significantly changed their expression levels. Of these miRNAs 16 were significantly differently expressed between gender. Using multi-variance analysis, nine miRNAs (let-7e-5p; miR-1; miR-19b-1-5p; miR-103a-3p; miR-148b-3p; miR-181b-5p; miR-188-5p; miR-423-5p; miR-874-3p) showed significantly different responses to maximal ergospirometry between genders., Conclusions: We report for the first time that in patients with CAD, miRNA expression is amenable to maximal ergospirometry and that the extent of changes differs between genders. Affected by exercise and gender were miRNAs that are associated, among others, with pathways for glucose metabolism, oxidative stress, and angiogenesis. Future studies should assess whether disease-specific miRNA expression in response to maximal exercise might serve as a marker for patient outcome., (© 2021 Walter de Gruyter GmbH, Berlin/Boston.)

Sedentary lifestyle predisposes to endothelial dysfunction, increased arterial stiffness and cardiovascular diseases, all of which can be positively modified by regular physical exercise training. A decrease in physical activity during winter months coincides with higher rates of cardiovascular events. In order to identify winter sports suitable to overcome this seasonal exercise deficit and thus contribute to cardiovascular health, it was the aim of this study to compare immediate effects of cross-country skiing (XCS) and alpine skiing (AS) on arterial stiffness as an alternative to indoor cycling (IC). After baseline assessment, eighteen healthy subjects performed one session of XCS, AS, and IC in randomized order. Pulse wave analysis was conducted (Mobil-o-Graph ® ) before and 10-min after exercise. Parameters of arterial stiffness and wave reflection were reduced after XCS and IC, but not after AS: central systolic blood pressure (IC: -8.0 ± 5.4 mmHg; p < 0.001), amplitude of the backward pressure wave (IC: -1.4 ± 2.7 mmHg; p < 0.05), reflection coefficient (XCS: -6.0 ± 7.8%; IC: -5.7 ± 8.1%; both p < 0.1), and pulse wave velocity (IC by -0.19 ± 0.27 m/s; p < 0.01). Higher exercise intensities correlated with greater reductions of arterial stiffness (all p < 0.05). Single sessions of XCS, IC but not AS led to comparable improvement in arterial stiffness, which was even more pronounced during higher exercise intensities. With regard to arterial stiffness, IC and XCS emerge as more effective to counteract the winter exercise deficit and thus the deleterious cardiovascular effects of a sedentary lifestyle., (© Journal of Sports Science and Medicine.)

Background: Mycoplasma genitalium is associated with genital discharge syndrome, but limited prevalence data are available in South Africa. The prevalence rates of M. genitalium infection and human immunodeficiency virus (HIV) coinfection were determined in urogenital specimens collected from male and female patients presenting with genital discharge syndrome to a primary health care center in Johannesburg, South Africa from 2007 through 2014., Methods: Genital specimens from 4731 patients were tested by a validated in-house multiplex real-time polymerase chain reaction assay for the detection of Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, and M. genitalium. Sera were tested for HIV infection using the Determine HIV 1/2 and Unigold assays., Results: The relative prevalence of M. genitalium in males and females was 8.9% and 10.6%, respectively. The prevalence of HIV infection in those infected with M. genitalium, without other sexually transmitted infections (STIs), was significantly higher than in those without M. genitalium infection (48.9% vs. 40.5%, P = 0.014). This significant difference in HIV seroprevalence was particularly observed among females in the study cohort., Conclusions: The relative prevalence of M. genitalium and its association with prevalent HIV among females with vaginal discharge syndrome (VDS) calls for further research on the potential role of M. genitalium in the transmission and acquisition of HIV.

Aims: Exercise is a trigger for acute coronary events especially in the untrained. Identifying subjects at risk remains a challenge. We set out to assess whether a distinct pattern of micro ribonucleic acids (miRNAs) expressed in response to an acute bout of all-out exercise might exist that would allow discrimination between health and disease., Methods: Twenty healthy subjects and 20 patients with coronary artery disease (CAD) performed an all-out cycle ergometry. Total RNA was extracted from blood drawn before and after exercise. Each blood sample was analysed for 187 target miRNAs by quantitative reverse transcription polymerase chain reaction., Results: At baseline, 18 miRNAs allowed discrimination between healthy subjects and CAD patients. In response to an acute all-out exercise in healthy subjects 51 miRNAs and in CAD patients 60 miRNAs were significantly modulated (all p < 0.05). Using logistic regression analysis, a unique pattern of pre-exercise miR-150-5p, post-exercise miR-101-3p, miR-141-3p and miR-200b-3p together with maximal oxygen uptake and maximal power corrected for bodyweight allowed discrimination between healthy subjects and CAD patients with an accuracy of 92.5%., Conclusion: In this most comprehensive analysis of exercise effects on circulating miRNAs to date we demonstrate for the first time that a distinct combination of miRNAs together with variables of exercise capacity allow robust discrimination between healthy subjects and CAD patients. We postulate that miRNAs may eventually serve as biomarkers to identify patients with CAD and possibly even those at risk of exercise-induced cardiac events.

Background: Extra-genital Neisseria gonorrhoeae and Chlamydia trachomatis infections are mostly asymptomatic, and important reservoir sites of infection as they often go undetected and may be more difficult to eradicate with recommended therapeutic regimens. Commercial nucleic acid amplification tests (NAATs) have not received regulatory approval for the detection of N. gonorrhoeae and C. trachomatis in extra-genital specimens. The HOLOGIC® APTIMA Combo2 assay for N. gonorrhoeae and C. trachomatis has performed well in evaluations using extra-genital specimens., Methods: We assessed the performance of an in-house real-time duplex PCR assay for the detection of N. gonorrhoeae and C. trachomatis in urine and extra-genital specimens using the HOLOGIC® APTIMA assays as gold standard comparators. Urine, oropharyngeal and ano-rectal specimens were collected from each of 200 men-who-have-sex-with-men (MSM) between December 2011 and July 2012., Results: For N. gonorrhoeae detection, the in-house PCR assay showed 98.5-100% correlation agreement with the APTIMA assays, depending on specimen type. Sensitivity for N. gonorrhoeae detection was 82.4% for ano-rectal specimens, 83.3% for oropharyngeal specimens, and 85.7% for urine; and specificity was 100% with all specimen types. The positive predictive value (PPV) for N. gonorrhoeae detection was 100% and the negative predictive value (NPV) varied with sample type, ranging from 98.5-99.5%. For C. trachomatis detection, correlation between the assays was 100% for all specimen types. The sensitivity, specificity, PPV and NPV of the in-house PCR assay was 100% for C. trachomatis detection, irrespective of specimen type., Conclusion: The in-house duplex real-time PCR assay showed acceptable performance characteristics in comparison with the APTIMA® assays for the detection of extra-genital N. gonorrhoeae and C. trachomatis.

Introduction: To counteract the winter activity deficit, we set out to analyze cardiorespiratory and metabolic responses of two high-intensity training (HIT) protocols during alpine skiing (AS), cross-country skiing (XCS), and indoor cycling (IC) and the effects of sex, age, and fitness level in this comparison., Methods: Nineteen healthy subjects (two age and fitness groups, both sexes) performed AS, XCS, and IC with measurements of oxygen uptake (V˙O2), energy expenditure (EE), HR, lactate, blood glucose and rate of perceived exertion, determined during 4 min of continuous HIT (HITc: 90% HRmax for XCS and IC or short turn skiing during AS) or 10-min intermittent HIT [HITint: 5 × 1 min high intensity (>90% HRmax or short turn skiing), 1 min active recovery]., Results: During all three exercise modes and irrespective of HIT protocols, sex, age, and fitness, participants were able to reach exercise intensities >90% HRmax and >84% V˙O2max. In all exercise modes 10-min of HITint with a 10-min postexercise O2 consumption phase resulted in greater mean EE per minute compared to 4-min HITc with 10 min postexercise O2 consumption. When applying the same HIT loading and recovery pattern to all three exercise modes, EE during approximately 1:15 h of AS was equivalent to about 1:00 h of either XCS or IC., Conclusions: Across all exercise modes and HIT protocols, high cardiorespiratory and metabolic responses were achieved regardless of age, sex, or fitness. EE during AS can be maximized by choosing the skiing mode "short turn skiing" in combination with an HITint to prolong the duration of continuous high-intensity loading during each descent. Therefore, all exercise modes and both HIT protocols are applicable and feasible in a broad spectrum of healthy subjects.

Background: We investigated the prevalence of human papillomavirus (HPV) infection and associated behavioural risk factors in men-who-have-sex-with-men (MSM) attending a clinical service in Cape Town, South Africa., Methods: MSM were enrolled at the Ivan Toms Centre for Men's Health in Cape Town. A psychosocial and sexual behavioral risk questionnaire was completed for each participant and urine, oro-pharyngeal and anal swabs were collected for HPV testing using the Linear Array HPV Genotyping Test. Logistic regression analyses were performed to determine sexual risk factors associated with HPV infection at the three anatomical sites., Results: The median age of all 200 participants was 32 years (IQR 26-39.5), of which 31.0 % were black, 31.5 % mixed race/coloured and 35.5 % white. The majority of the participants (73.0 %) had completed high school, 42.0 % had a tertiary level qualification and 69.0 % were employed. HPV genotypes were detected in 72.8 % [95 % CI: 65.9-79.0 %], 11.5 % [95 % CI: 7.4-16.8 %] and 15.3 % [95 % CI: 10.5-21.2 %] of anal, oro-pharyngeal and urine specimens, respectively. Prevalence of high-risk (HR)-HPV types was 57.6 % [95 % CI: 50.3-64.7 %] in anal samples, 7.5 % [95 % CI: 4.3-12.1 %] in oro-pharyngeal samples and 7.9 % [95 % CI: 4.5-12.7 %] in urine, with HPV-16 being the most common HR-HPV type detected at all sites. HPV-6/11/16/18 was detected in 40.3 % [95 % CI: 33.3-47.6 %], 4.5 % [95 % CI: 2.1-8.4 %] and 3.2 % [95 % CI: 1.2-6.8 %] of anal, oro-pharyngeal and urine samples, respectively. Multiple HPV types were more common in the anal canal of MSM while single HPV types constituted the majority of HPV infections in the oropharynx and urine. Among the 88 MSM (44.0 %) that were HIV positive, 91.8 % [95 % CI: 83.8-96.6 %] had an anal HPV infection, 81.2 % [95 % CI: 71.2-88.8 %] had anal HR-HPV and 85.9 % [95 % CI: 76.6-92.5 %] had multiple anal HPV types. Having sex with men only, engaging in group sex in lifetime, living with HIV and practising receptive anal intercourse were the only factors independently associated with having any anal HPV infection., Conclusions: Anal HPV infections were common among MSM in Cape Town with the highest HPV burden among HIV co-infected MSM, men who have sex with men only and those that practiced receptive anal intercourse. Behavioural intervention strategies and the possible roll-out of HPV vaccines among all boys are urgently needed to address the high prevalence of HPV and HIV co-infections among MSM in South Africa.

Objective: Endothelial dysfunction occurs early during atherogenesis and it can be normalized by exercise training. Unfortunately, patients' compliance with exercise prescription remains low, often because the given choices do not appeal to them. In Alpine regions, skiing is a popular mode of exercise, and therefore we set out to assess whether it can induce antiatherogenic effects., Methods: We randomized 42 subjects into a group of 12weeks of guided skiing (intervention group, IG, n=22; 12 males/10 females; age: 66.6±2.1years) or a control group (CG, n=20; 10 males/10 females; age: 67.3±4.4years). Early (CD3-CD34+CD45+) and late endothelial progenitor cells (EPCs; CD45dimCD34+KDR+ peripheral blood mononuclear cells, PBMCs), peripheral arterial tonometry and endothelial biomarkers were assessed at the beginning and end of the study., Results: In the IG, participants completed 28.5±2.6 skiing days at an average heart rate of 72.7±8.5% of their maximum heart rate. Changes in early (IG: +0.001±0.001% PBMC; CG: -0.001±0.001% PBMC; IG vs. CG: p<0.001) but not late EPCs differed significantly. Changes in peripheral arterial tone differed significantly between IG (Reactive Hyperemia Index: +0.18±0.76) and CG (-0.39±0.85; p=0.045), as did homocysteine (IG: -1.3±1.3μmol/l; CG: -0.4±1.4μmol/l; p=0.037) while other endothelial biomarkers remained essentially unchanged., Conclusions: This study shows that skiing induces several beneficial effects on markers of atherogenesis including EPCs, peripheral arterial tone and homocysteine. Our findings suggest that recreational alpine skiing may serve as a further mode of preventive exercise training, which might result in improved compliance with current recommendations., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Since physical inactivity especially prevails during winter months, we set out to identify outdoor alternatives to indoor cycling (IC) by comparing the metabolic and cardiorespiratory responses during alpine skiing (AS), cross-country skiing (XCS) and IC and analyse the effects of sex, age and fitness level in this comparison. Twenty one healthy subjects performed alpine skiing (AS), cross-country skiing (XCS), and IC. Oxygen uptake (VO2), total energy expenditure (EE), heart rate (HR), lactate, blood glucose and rate of perceived exertion (RPE) were determined during three 4-min stages of low, moderate and high intensity. During XCS and IC VO2max and EE were higher than during AS. At least 2½ hours of AS are necessary to reach the same EE as during one hour of XCS or IC. HR, VO2, lactate, and RPEarms were highest during XCS, whereas RPEwhole-body was similar and RPElegs lower than during AS and IC, respectively. Weight adjusted VO2 and EE were higher in men than in women while fitness level had no effect. Male, fit and young participants were able to increase their EE and VO2 values more pronounced. Both AS and XCS can be individually tailored to serve as alternatives to IC and may thus help to overcome the winter activity deficit. XCS was found to be the most effective activity for generating a high EE and VO2 while AS was the most demanding activity for the legs. Key pointsDuring cross-country skiing and indoor cycling VO2max and energy expenditure were higher than during alpine skiingApproximately 2½ hours of alpine skiing are necessary to reach the same energy expenditure of one hour of cross-country skiing or indoor cycling.Alpine skiing and cross-country skiing can be individually tailored to serve as sports alternatives in winter to activity deficit.By applying different skiing modes as parallel ski steering, carving long radii and short turn skiing, metabolic and cardiorespiratory response can be increased during alpine skiing.Male, fit and young participants were able to increase their energy expenditure and VO2 more pronounced with an increase in intensity compared with their counterparts.

Background: Mycoplasma genitalium is a common sexually transmitted infection associated with human immunodeficiency virus (HIV) infection. Some studies suggest that M. genitalium may increase the risk of HIV acquisition. However, results have been inconsistent, and this association has never been examined longitudinally., Methods: Stored endocervical samples from a longitudinal cohort study of 131 Zimbabwean women in whom HIV-1 seroconversion recently occurred were tested for detection and quantity of M. genitalium using polymerase chain reaction analysis. The associations between M. genitalium and the detection and quantity of genital HIV type 1 (HIV-1) RNA, the detection and quantity of plasma HIV-1 RNA, and the CD4(+) T-cell count was analyzed using mixed-effects regression analysis., Results: M. genitalium was detected in 10.5% of stored specimens (44 of 420), and infection persisted for up to 300 days. M. genitalium was independently associated with detection of genital HIV-1 RNA (adjusted odds ratio, 2.67; 95% confidence interval, .99-7.20), after adjustment for plasma viral load, viral set point, CD4(+) T-cell count, herpes simplex virus type 2 detection, and gonorrhea. There was no evidence of an association between M. genitalium detection or quantity and either plasma HIV-1 RNA load or CD4(+) T-cell count., Conclusions: The growing evidence for an association between M. genitalium and HIV genital shedding and the high prevalence and persistence of M. genitalium in this population suggest that further research into this association is important. Consideration of the cost-effectiveness of M. genitalium screening interventions may be warranted., (© The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)

A real-time PCR (RT-PCR) assay was modified to simultaneously detect Neisseria gonorrhoeae and to determine gonococcal susceptibility to ciprofloxacin using clinical samples. The modified RT-PCR assay was validated using DNA extracted from 40 linked isolates and urethral swabs, 24 of which had linked first-pass urine samples, obtained from men presenting with urethral gonorrhoea. The RT-PCR assay enabled amplification of N. gonorrhoeae dcmH, gyrA and parC genes. The quinolone resistance-determining regions (QRDRs) of the isolates' gyrA and parC genes were sequenced. Following successful validation, 33 first-pass urine-derived DNA extracts, obtained from men with gonorrhoea, were tested with the assay and results were compared with blinded ciprofloxacin susceptibility data. Gonococcal susceptibility to ciprofloxacin correlated perfectly with gyrA amplicon generation. No gyrA amplicons were detected for gonococcal infections due to ciprofloxacin-intermediate/resistant organisms. Amplification of parC correlated less well with ciprofloxacin susceptibility phenotypes. Simultaneous non-generation of gyrA and parC amplicons consistently predicted the presence of ciprofloxacin-resistant gonococci. Characteristic point mutations in the gyrA/parC QRDRs were found in DNA amplified from those extracts that failed to produce gyrA/parC amplicons. The RT-PCR assay performed well with DNA extracted from first-pass urine specimens and results correlated perfectly with ciprofloxacin susceptibility phenotypes. In conclusion, the modified RT-PCR assay can detect N. gonorrhoeae in DNA extracted from first-pass urine specimens of men with urethral gonorrhoea and accurately predicts gonococcal susceptibility to ciprofloxacin. This molecular assay provides a useful tool for surveillance and patient management in settings where fluoroquinolones can still be used for treatment of gonorrhoea., (Copyright © 2013 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.)

Objectives: To describe the phenotypic and genetic characteristics of the first two cases of extended-spectrum cephalosporin (ESC)-resistant Neisseria gonorrhoeae in South Africa, one of which was associated with verified cefixime treatment failure., Patients and Methods: Two ESC-resistant N. gonorrhoeae isolates were cultured from the urethral discharge of two men who have sex with men (MSM). One man reported a persistent urethral discharge that had failed to respond to previous therapy with oral cefixime. Agar dilution MICs were determined for eight antibiotics. β-Lactam-associated resistance mutations were identified through PCR-based amplification and sequencing for several key genes: penA, mtrR and its promoter, porB1b (penB), ponA and pilQ. For molecular epidemiological characterization, full-length porB gene sequencing, N. gonorrhoeae multiantigen sequence typing (NG-MAST) and multilocus sequence typing (MLST) were performed., Results: Both isolates were resistant to cefixime, ciprofloxacin, penicillin and tetracycline and intermediate/resistant to azithromycin, but susceptible to ceftriaxone, gentamicin and spectinomycin. Both isolates had the type XXXIV penA mosaic allele in addition to previously described resistance mutations in the mtrR promoter (A deletion), porB1b (penB) (G101K and A102N) and ponA1 (L421P). Both isolates had an identical NG-MAST sequence type (ST4822) and MLST sequence type (ST1901)., Conclusions: Both isolates were resistant to cefixime and possessed a number of identical mutations in key genes contributing to ESC resistance in N. gonorrhoeae. The two isolates contained the type XXXIV penA mosaic allele and belonged to a successful international MSM-linked multidrug-resistant gonococcal clone (MLST ST1901) associated with several cefixime treatment failures in Europe and North America.

Background: The abrupt fall in estrogens levels during the menopausal transition may connote an hormonal state predisposing to neurodegenerative disorders, e.g. Alzheimer's disease (AD). Reportedly, the neurotrophic activity of estrogen involves an interaction with IGF-I., Aim: To evaluate the leukocyte gene expression of progesterone receptor (PR-A/B) and interleukin 6 (IL-6), two parameters under the control of estrogens and involved in the pathogenesis of AD., Subjects: The study was conducted in non-demented women divided into two groups according to their pre- or post-menopausal state; each group being further divided into two subgroups based on their circulating levels of IGF-I (normal or low). An additional sample of AD-affected women served as a comparison group., Results: Estrogens maintained their full activity only when IGF-I levels were in the range of normalcy. On the contrary, if the concentrations of one or both hormones were reduced, estrogens were not anymore capable to control the gene expression of PR-A/B or IL-6., Conclusions: Before administering hormone-based replacement therapy, characterization of the somatotropic function should be performed in the early phase of the menopause.

Objective: Changes in circulating levels of many adipocyte-derived peptides, including adipokines such as adiponectin, leptin and tumor necrosis factor alpha (TNF-α), have been reported in obesity (OB). Somatostatin (SRIF) inhibits circulating levels of adiponectin and leptin in lean (LN) subjects, but the effect of a SRIF infusion on these adipokines, including TNF-α, in OB is to date unknown., Methods: Ten young women (5 OB and 5 LN) were studied. All subjects underwent an infusion of SRIF (9 μg/kg/h i.v., over 60 min), with blood samples drawn prior to and at different time intervals after SRIF administration. Plasma levels of adiponectin, leptin and TNF-α were measured at each interval., Results: Basal levels of leptin and TNF-α were significantly higher in OB than LN women, whereas levels of adiponectin were significantly lower in OB than LN subjects. SRIF significantly inhibited plasma concentrations of adiponectin (at 60 min) in both OB and LN women, without affecting those of leptin and TNF-α in either group. In LN subjects, the inhibitory effect of SRIF on plasma adiponectin persisted up to 150 min, whereas SRIF infusion withdrawal in OB women resulted in a prompt restoration of basal levels of the adipokine., Conclusions: Plasma concentrations of leptin and TNF-α, which are higher in OB than LN subjects, are unaffected by a SRIF infusion, which, in contrast, inhibits circulating levels of adiponectin in both groups, with a delayed return to the baseline secretion of the adipokine in LN subjects., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Human GH (hGH) is a heterogeneous protein hormone consisting of several isoforms. This heterogeneity is the consequence of multiple hGH genes, mRNA splicing, post-translational modifications, and peripheral metabolism, and it represents one important reason for the disparity among GH assay results from different laboratories. However, other factors are involved: a) interference from endogenous GH binding proteins; b) different specificities of anti- GH (monoclonal and polyclonal) antibodies; c) different matrix effects among the calibrators; d) the use of different calibrators. The measurement of GH levels in response to provocative testing is an essential part of the diagnosis of GH deficiency. For this purpose, an accurate, reproducible and universally valid GH measurement would be highly desirable, but, despite a huge number of efforts in clinical biochemistry, this goal remains elusive.

Objectives: To determine whether the 23S ribosomal RNA (rRNA) A2058G and A2059G mutations that confer macrolide resistance are present among southern African strains of Treponema pallidum and to determine their subtype distribution., Methods: 117 genital ulcer specimens, collected between March 2005 and April 2010 in South Africa and Lesotho and previously determined to be positive for T pallidum DNA by molecular testing, were retested using a commercial real-time PCR assay. Those specimens that were still positive for T pallidum DNA were screened for the macrolide resistance-encoding point mutations in the 23S rRNA gene using rapid PCR-based restriction digest assays. Molecular characterisation of two variable treponemal genes, arp and tpr, was used to subtype the T pallidum strains., Results: 1 of 100 T pallidum-positive specimens, collected in Lesotho, contained the A2058G macrolide resistance-encoding 23S rRNA gene mutation, whereas the A2059G mutation was absent. It was possible to fully type 97/100 of all T pallidum DNA-positive samples. A total of nine arp repeat sizes, nine tpr patterns and a combined total of 20 subtypes were identified. Overall, the most common subtypes were 14d (32%), followed by 17d (12%), 14a (10%), 14b (8%), 22b (6%) and 14i (5%). Subtypes 14d and 14a were the predominant subtypes in samples from South Africa (43%) and Lesotho (22%), respectively., Conclusions: Macrolide resistance among T pallidum strains appears to be uncommon in southern Africa. Although a high degree of genetic heterogeneity was observed among the strains tested, T pallidum subtype 14d appears to be the predominant circulating strain.

We developed and validated a real-time quantitative polymerase chain reaction (qPCR) assay to determine Mycoplasma genitalium bacterial load in endocervical swabs, based on amplification of the pdhD gene which encodes dihydrolipoamide dehydrogenase, using the Rotor-Gene platform. We first determined the qPCR assay sensitivity, limit of detection, reproducibility and specificity, and then determined the ability of the qPCR assay to quantify M. genitalium in stored endocervical specimens collected from Zimbabwean women participating in clinical research undertaken between 1999 and 2007. The qPCR assay had a detection limit of 300 genome copies/mL and demonstrated low intra- and inter-assay variability. The assay was specific for M. genitalium DNA and did not amplify the DNA from other mycoplasma and ureaplasma species. We quantified M. genitalium in 119 of 1600 endocervical swabs that tested positive for M. genitalium using the commercial Sacace M. genitalium real-time PCR, as well as 156 randomly selected swabs that were negative for M. genitalium by the same assay. The M. genitalium loads ranged between <300 and 3,240,000 copies/mL. Overall, the qPCR assay demonstrated good range of detection, reproducibility and specificity and can be used for both qualitative and quantitative analyses of M. genitalium in endocervical specimens and potentially other genital specimens., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Objective: Changes in many gastrointestinal peptides, including the anorexigenic peptide YY (PYY), which is produced by L cells, occur in both anorexia nervosa (AN) and obesity (OB). High PYY levels are present in AN, whereas in morbid OB fasting and postprandial PYY secretion is blunted. Somatostatin (somatotropin release-inhibiting factor (SRIF)) reportedly inhibits plasma PYY concentrations in animals and healthy humans, but the effect of a SRIF infusion on spontaneous PYY secretion in AN and OB is unknown., Methods: A total of 18 young women, seven with acute AN (A-AN), four with AN in the recovery phase (R-AN), and seven with morbid OB, were studied. All subjects underwent an infusion of SRIF (9 μg/kg i.v./h, over 60 min), with blood samples drawn before and at different time intervals after SRIF administration. Plasma PYY levels were measured at each time point., Results: SRIF significantly inhibited plasma PYY concentrations in R-AN and OB, without affecting PYY titers in A-AN. In OB, the inhibitory effect of SRIF also persisted at 90 min. Withdrawal of SRIF infusion in R-AN resulted in a prompt restoration of basal plasma PYY levels, whereas termination of SRIF infusion in OB was followed by a slower increase of PYY titers toward baseline levels. After infusion, PYY Δ area under the curve (ΔAUC) in R-AN was significantly higher than those in A-AN and OB patients. A significant difference in PYY ΔAUC between A-AN and OB was present., Conclusions: These results suggest the existence of a hypo- and hyper-sensitivity of L cells to the inhibitory effect of SRIF in A-AN and OB respectively.

Objectives: To determine the complete nucleotide sequence of the novel Johannesburg β-lactamase-encoding gonococcal plasmid (pEM1) and to determine the strain relatedness of Johannesburg plasmid-containing penicillinase-producing Neisseria gonorrhoeae (PPNG) by molecular typing., Methods: Eleven PPNG isolates containing the Johannesburg β-lactamase-encoding plasmid were previously identified among gonococci isolated from men with urethral discharge attending a clinic in Alexandra (Johannesburg) using a PCR assay. DNA sequence-based characterization of one such plasmid was performed to determine its relatedness to the prototype Asia plasmid. The 11 PPNG isolates containing the Johannesburg plasmid and 105 other clinical gonococci isolates were typed using N. gonorrhoeae multi-antigen sequence typing (NG-MAST)., Results: Plasmid pEM1 was determined to comprise 4865 bp and to be a deletion derivative of the prototype Asia plasmid with a unique 2560 bp deletion in the non-TnA region. NG-MAST genotyping demonstrated a significant association between sequence type (ST) 502, or other closely related STs, and the Johannesburg plasmid-containing PPNG (P < 0.0001)., Conclusions: Sequencing of a novel β-lactamase-encoding plasmid (pEM1) found in PPNG isolates in Johannesburg shows it to be a deletion derivative of the prototype Asia plasmid, the deletion most likely arising as a result of DNA rearrangements. The majority of Johannesburg plasmid-containing PPNG isolates were, or were very closely related to, ST502.

Obese patients have decreased fasting and postprandial levels of peptide YY (PYY), an anorexigenic peptide produced by the L cells of the gastrointestinal mucosa. Fatty nutrients are the most powerful stimulus for PYY release. Cholestyramine, an anion exchanger which adsorbs bile salts, reduces digestion of lipids. The aim of the present study was to investigate the effects of cholestyramine or placebo on PYY secretion in obese women administered a high-fat meal [n=8; age: 30.9±2.7 years; BMI: 47.3±3.3 kg/m2]. Postprandial PYY levels in obese women given placebo significantly increased in plasma at 30, 60, 90, and 120 min after meal ingestion. Cholestyramine administration significantly reduced postprandial PYY response at 15, 30, and 60 min. Percent fat mass (FM%) was negatively correlated with the percent increment of plasma PYY concentrations induced by meal administration at 30 min; conversely, there was a positive correlation between FM% and the percent decrement of plasma PYY concentrations induced by cholestyramine at the same time interval. These correlations failed to reach statistical significance when related to BMI. This study implies that in the obese state the altered PYY response to food consumption is a consequence of a dysfunction of L cells, which become less sensitive to the positive feedback effect of lipids., (Georg Thieme Verlag KG Stuttgart · NewYork.)

Background: To detect and type plasmids responsible for penicillin and tetracycline resistance in Neisseria gonorrhoeae isolates using a novel duplex polymerase chain reaction (PCR) assay., Methods: A duplex PCR assay, to detect and type penicillinase-producing N. gonorrhoeae (PPNG), and plasmid-mediated tetracycline resistant N. gonorrhoeae (TRNG), was developed on the basis of published single assays. Gonococcal Isolate Surveillance Project control strains were used in assay development and then 209 consecutive N. gonorrhoeae isolates, collected from men with urethral discharge in 2008, were tested. Controls included Asia, Africa, and Toronto β-lactamase plasmids, as well as American and Dutch TRNG plasmids. PCR amplicons were detected using an Agilent 2100 Bioanalyzer. Minimum inhibitory concentrations (MIC) were determined with E tests. Penicillinase production was detected using Nitrocefin solution., Results: Among 209 gonococcal isolates, 54 (25.8%) PPNG and 154 (73.3%) TRNG were detected. The MIC50 and MIC90 values were determined for penicillin (0.19 and 32 mg/L) and tetracycline (6 and 16 mg/L). The assay detected the Africa-type (35.2%), the Toronto-type (44.4%), and a new type (20.3%) of β-lactamase plasmid. The American-type TRNG plasmid was 3-fold more frequent as compared with the Dutch-type. Although there was no overall association between the detection of PPNG and TRNG plasmids, only American type TRNG contained β-lactamase-encoding plasmids (P < 0.0001)., Conclusions: The prevalence of plasmid-mediated resistance to tetracycline, and to a lesser extent penicillin, is high and neither drug is likely to have any future role in the treatment of gonorrhoea in South Africa. A novel β-lactamase plasmid was detected during the study and requires further characterization.

Background: This study was undertaken to establish the etiology of the male urethral discharge (MUDS) and vaginal discharge (VDS) syndromes, to determine the prevalence of other sexually transmitted infections (STI) and human immunodeficiency virus (HIV) coinfections, and to examine associations between STIs and HIV serostatus among STI patients in South Africa., Methods: A total of 507 MUDS and 300 VDS patients were recruited in Cape Town (CPT) and Johannesburg (JHB). A multiplex polymerase chain reaction assay detected Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, and Mycoplasma genitalium infections. Bacterial vaginosis and candidiasis were detected by microscopy. Sera were screened for syphilis, HSV-2, and HIV antibodies., Results: Etiological diagnoses were made for 92% of MUDS patients and 85% of VDS patients. Gonorrhoea accounted for 85% (CPT) and 71% (JHB) of MUDS presentations. Chlamydia was the second most frequently detected MUDS pathogen (CPT, 13%; JHB, 24%). Among VDS patients, bacterial vaginosis was the most common cause (CPT, 46%; JHB, 36%) and trichomoniasis the most frequently detected STI pathogen (CPT, 19%; JHB, 34%). Few patients (4%) had serological evidence of syphilis. The HSV-2 and HIV seroprevalence were higher in Johannesburg compared to Cape Town and among women compared to men. HIV infection was statistically significantly associated with HSV-2 seropositivity at both sites and with the presence of N. gonorrhoeae and absence of C. trachomatis in Cape Town MUDS patients., Conclusions: Gonorrhoea and bacterial vaginosis were confirmed as the most frequent causes of MUDS and VDS. The high HIV seroprevalence in STI patients emphasizes the need to address HIV testing among this population.

Objectives: To investigate the distribution of human papillomavirus (HPV) genotypes and determine the associations between HPV infection and HIV coinfection in sexually active heterosexual men with anogenital warts (GW), male urethral discharge or asymptomatic men., Methods: Valid specimens for HPV genotyping were obtained from three patient groups consisting of 108 men with GW, 56 men with urethral discharge syndrome and 50 asymptomatic men attending for HIV voluntary counselling and testing. The Linear Array HPV Genotyping Test was used to determine the HPV genotype distribution among study participants. Sera were tested for HIV antibodies using two commercial rapid tests., Results: The prevalence of anogenital HPV among study participants was 78% (166). HPV DNA was detected in 100% (108) of GW, 48% (27) of men with urethral discharge syndrome and 62% (31) of voluntary counselling and testing participants. HPV types 6, 11, 16 and 18 were prevalent as either single or combined infections in 81% (134) of all HPV-positive study participants. HPV types 6 and/or 11 were significantly higher among GW patients (p<0.001). After adjusting for patient groups, HIV seropositivity was significantly associated with multiple HPV infections (OR=3.98, 95% CI 1.58 to 10.03) but not with the presence of a foreskin (OR=0.67, 95% CI 0.32 to 1.40)., Conclusions: Infections with HPV were prevalent among sexually active heterosexual men attending the men's sexual health clinic. Associations were observed between HIV coinfection and multiple HPV infections. Further population-based studies on the prevalence of HPV genotypes are required to determine if men should be included in any future national HPV vaccination programme in South Africa.

Background and Aims: Ghrelin is an orexigenic 28-amino acid peptide produced by the stomach. Circulating ghrelin levels rise shortly before and fall shortly after every meal. Peptide YY (PYY), an anorexigenic 36-amino acid peptide, is secreted primarily from the intestinal mucosa of the ileum and large intestine. Plasma PYY levels begin to rise within 15 min after starting to eat and plateau within approximately 90 min, remaining elevated for up to 6 h. Recently, some studies have tried to evaluate the potential role of ghrelin and PYY in the hyperphagia of patients with Prader-Willi syndrome (PWS). While hyperghrelinemia is well characterized in PWS, conflicting results have been reported for PYY. The aim of the study was to investigate ghrelin and PYY responses to a standard liquid high-fat meal in children with PWS., Patients and Methods: Circulating levels of total ghrelin and PYY levels were assayed by RIA after overnight fasting and 45, 60, 90, and 180 min following a standard meal (Ensure 6 ml/kg) in 16 patients with PWS (11 boys and five girls, aged 4.6-10.7 years, including ten receiving 0.02 mg/kg per day rhGH for 2-18 months; body mass index (BMI) z-score: 0.6+/-0.2 and 1.6+/-0.5 for children treated or not treated with rhGH respectively), ten obese (eight boys and two girls, aged 9.2-15.6 years; BMI z-score: 2.4+/-0.2, i.e. BMI >97th centile for chronological age and sex) subjects, and 16 normal-weight controls (five boys and 11 girls, aged 5.8-17.3 years; BMI z-score: 0.6+/-0.2)., Results: PWS children showed higher fasting levels of ghrelin than obese and lean controls. Postprandial ghrelin drop was more pronounced in PWS than in the other study groups. No significant difference on fasting levels of PYY was found among groups. PWS showed a higher postprandial PYY rise than obese and lean controls. PWS patients treated and not treated with GH showed similar fasting and postprandial levels of ghrelin and PYY. Fasting PYY levels correlated negatively (P<0.05; r=-0.68) with those of ghrelin only in PWS., Conclusions: The results of this study confirm fasting hyperghrelinemia in PWS. Since in PWS adults an impaired postprandial suppression of plasma ghrelin was previously reported to be associated with a blunted postprandial PYY response, the finding of a meal-induced decrease and increase in ghrelin and PYY levels respectively in PWS children would imply that the regulation of appetite/satiety of these peptides is operative during childhood, and it progressively deteriorates and vanishes in adulthood when hyperphagia and obesity worsen.

Objective: To detect exogenous recombinant human GH (rhGH) abuse in female athletes., Design: GH-dependent markers were assayed in serum of 100 female athletes (control group) and in a subgroup of nine female subjects treated with rhGH (0.09 IU/kg body weight, 6 days/week for 3 weeks)., Methods: Cut-off values (mean+2 s.d.) for IGF1, N-terminal propeptide of type III procollagen (PIIINP) and C-terminal telopeptide of type I collagen (ICTP) were calculated and arbitrary scores (1.5 or 2.0) were assigned to abnormal markers. By using the sum of individual marker scores, positive (> or =3) or negative (<3) scores were obtained., Results: None of the control group obtained a positive score (> or =3). Abnormal IGF1, PIIINP and ICTP levels were found in 61.4, 54.5 and 11.4% samples of the treated group. Overall, positive cases were present in 43.2% blood samples drawn in subjects treated with rhGH and in 26% of samples after rhGH withdrawal. The sensitivity of the detection approach was 66.6% at the end of 3-week rhGH treatment and 11.1% at the 15th day of rhGH withdrawal, while the specificity was 100%., Conclusion: Detection test for rhGH administration appears less sensitive in female (66.6%) than in male athletes (previous observation, 100% after 3 weeks of comparable rhGH dose), but shows a similar specificity (98.5-100%). Since athletes supposedly use very high doses and long-term administration of rhGH for doping purposes, it is foreseen that the here-in detection test would in future increase its strength.

Background and Objective: Incidence and prevalence of Alzheimer's disease (AD) are higher in postmenopausal women than in age-matched men. Since at menopause the endocrine system and other biological paradigms undergo substantial changes, we thought to be of interest studying whether (and how) the balance between some biological parameters allegedly neuroprotective (e.g. related to estrogen, dehydroepiandrosterone and CD36 functions) and others considered pro-neurotoxic (e.g. related to glucocorticoid and interleukin-6 activities) vary during lifespan in either sex in either normalcy or neurodegenerative disorders., Subjects and Methods: Along with this aim, we evaluated the gene expression levels of estrogen receptors (ERs), glucocorticoid receptors (HGRs), interleukin-6 (IL-6) and CD36, a scavenger receptor of class B allegedly playing a key role in the proinflammatory events associated with AD, in a population of 209 healthy subjects (73M, 106F, 20-91-year old) and 85 AD patients (36M, 49F, 65-89-year old). Results obtained were related to plasma titers of estrogens, cortisol and dehydroepiandrosterone sulfate (DHEAS). Studies were performed in peripheral leukocytes, since these cells (1) are easily obtainable by a simple blood sampling, (2) express many molecules and multiple receptors which are under the same regulatory mechanisms as those operative in the brain and (3) some of them, e.g. monocytes, share many functions with microglial cells., Results: In healthy men all the study parameters were quite stable during lifespan. In women, instead, at menopausal transition, some changes that may predispose to neurodegeneration occurred. In particular, there was (1) an up-regulation of ERs, and a concomitant increase of IL-6 gene expression, events likely due to the loss of the inhibitory control exerted by estradiol (E(2)); (2) an increase of HGR alpha:HGR beta ratio, indicative of an augmented cortisol activity on HGR alpha not sufficiently counteracted by the inhibitory HGR beta function; (3) a reduced CD36 expression, directly related to the increased cortisol activity; and (4) an augmented plasma cortisol:DHEAS ratio, widely recognized as an unfavorable prognostic index for the risk of neurodegeneration. In AD patients of both sexes, the expression of the study parameters was similar to that found in sex- and age-matched healthy subjects, thus indicating their unrelatedness to the disease, and rather a better correlation with biological events., Conclusions: Menopausal transition is a critical phase of women's life where the occurrence of an unfavorable biological milieu would predispose to an increased risk of neurodegeneration. Collectively, the higher prevalence of AD in the female population would depend, at least in part, on the presence of favoring biological risk factors, whose contribution to the development of the disease occurs only in the presence of possible age-dependent triggers, such as beta-amyloid deposition.

Myostatin and mechano-growth factor (MGF), an isoform of insulin-like growth factor-I (IGF-I), are two important regulators of muscle hypertrophy. The aim of the present study was to investigate the effects of recombinant human growth hormone (rhGH) and/or testosterone on muscle MGF/IGF-IEa/myostatin expression in intact and hypophysectomized rats treated for 15 d with 1) saline or rhGH, 2) sesame oil or testosterone, 3) saline+sesame oil, or rhGH+testosterone (first experiment) or for 7 d with saline or rhGH (second experiment). Animals were killed by decapitation 24 h or 4 d after the last injection (first or second experiment, respectively). Muscle expressions of MGF, IGF-IEa, and myostatin were determined by RT-PCR. A significant increase in the weight of gastrocnemius muscle was observed only in hypophysectomized rats treated with rhGH alone or in combination with testosterone. Administration of rhGH to hypophysectomized rats caused a marked increase in both MGF and IGF-IEa muscle mRNA levels (without any change in the muscle expression of myostatin), an effect that was abolished when testosterone was combined with rhGH. Conversely, in intact rats rhGH increased myostatin muscle mRNA levels without affecting those of MGF and IGF-IEa. Testosterone, alone or combined with rhGH, induced an inhibition of myostatin expression in the muscle of intact rats, but did not change muscle paradigms of hypophysectomized rats. In conclusion, rhGH and/or testosterone anabolic effects in the muscle are mediated by a different expression of MGF/IGF-IEa/myostatin, which is related to the pituitary function.

Many studies have been prompted in the last few years aimed at clarifying and disentangle the different components involved in the control of metabolism and body weight. Based on a host of neuroanatomic, immunohistochemical and physiological findings, it is now clear that the hypothalamus is activated or deactivated by a series of afferent signals either hormonal or neural originated by peripheral tissues, which reflect changes in the global energy state. These hormonal and neural signals mainly stem from the adipose tissue, which is now considered a true endocrine organ, and, alternatively, by the gastrointestinal tract, in relation to fasting, feeding or refeeding conditions, and are vehicled to the brain where they act on receptors of an increasing family of first and second order peptides. This contribution at first presents the key elements of the complex mechanisms of control of metabolism and body weight, and then reports on recent presented advances in neuroendocrine and autonomic regulation of metabolic functions.

CD36, a scavenger receptor of class B (SR-B), helps mediate microglial and macrophage response to beta-amyloid fibrils (betaA), and seems to play a key role in the proinflammatory events associated with Alzheimer disease (AD) in many tissues. Peripheral leukocytes express many molecules and multiple receptors which undergo the same regulatory mechanisms as those operative in the brain. Thus, these cells, easily obtainable through peripheral blood sampling, may be used as a tool to investigate changes occurring in inaccessible brain areas. Based on these premises, we investigated the leukocyte expression of CD36 in 70 AD patients and in 30 subjects with mild cognitive impairment (MCI). Results were compared to those of 20 young and 40 age-matched control subjects. Leukocyte expression of CD36 was significantly reduced versus controls in both AD and MCI patients, while in young and old controls there were no age-related changes. Although preliminary, these data indicate that the reduction of CD36 expression in leukocytes is a disease-related phenomenon, occurring since the early stages of AD (MCI). Irrespective of the mechanism(s) underlying such changes, assessment of leukocyte CD36 expression might represent an useful tool to support the diagnosis of AD and to screen MCI patients candidates to develop the disease.

Acute respiratory distress syndrome (ARDS) and pneumonia are closely correlated in the critically ill patient. Whereas ARDS is often complicated by nosocomial pneumonia, pulmonary infection is also the most frequent single cause of ARDS. The prevalence of pneumonia during the course of ARDS seems to be particularly high, but whether persons with ARDS are more susceptible to pneumonia or simply have more risk factors remains unknown because of methodological limitations. Recent research suggests that host factors have a major bearing on the development of ARDS. To date, sepsis seems to be the principal link between pneumonia and ARDS. However, prospective observational data on this supposed sequence are not available. The individual role of specific pathogens for the development of ARDS is difficult to assess, because prospective studies are missing. Respiratory viruses have received particular attention, but this review suggests that infections with coronavirus and avian influenza virus (H5N1) are associated with a high incidence of ARDS.

The cannabinoid CB1 receptor antagonist SR141716 (Rimonabant) is known to reduce food intake by central and peripheral mechanisms. Recently, SR141716 has been reported to block the orexigenic effect of ghrelin, a potent orexigenic peptide produced by the stomach. This study investigated whether in rats, made tolerant to the hypophagic effect of SR141716, the drug was still capable to block the orexigenic activity of another non-natural (hypothalamic) peptide, i.e., the growth hormone releasing peptide (GHRP) hexarelin, a ghrelin mimetic. In the acute experiments, each dose of SR141716 (1, 5 and 10 mg/kg i.p.) reduced food intake with respect to vehicle-treated rats, whereas hexarelin (160 microg/kg s.c.) markedly stimulated feeding. All doses of SR141716 were capable to reduce the orexigenic effect of the GHRP. A 15-day administration of SR141716 (10 mg/kg i.p.) reduced both food intake and body weight. Tolerance to the hypophagic effect of SR141716 developed within 5 days, but in contrast, body weight remained markedly below that of vehicle-treated group throughout the entire treatment period. Interestingly, despite development of tolerance to its hypophagic effect, SR141716 was capable to suppress the orexigenic effect of repeated hexarelin challenge tests performed throughout the chronic experiments. In conclusion, the results of the present study confirm and broaden the existence of a functional relationship between ghrelin and endocannabinoids in the control of food intake, and bespeak the ability of a CB1 receptor antagonist to suppress orexia caused by stimuli alien to direct stimulation of the cannabinoid system.

Hypothalamic neurochemical alterations in mammals underlie disturbances of food intake. There is scarce information on these topics in elderly persons; therefore, the aims of the present study were: (i) to evaluate the orexigenic effects of a growth hormone secretagogue, administered to young and old rats and dogs, alone or in combination with molsidomine, a donor of nitric oxide and (ii) to evaluate by reverse transcription-polymerase chain reaction in the whole hypothalamus of young and old rats messenger RNA levels of a wide number of anabolic and catabolic peptides, receptors, and enzymes involved in the control of feeding behavior, relating the detected titers, whenever possible, to the feeding responses to growth hormone secretagogue. In all, the results obtained strengthen the proposition that, in the hypothalamus of old rats, anti-anorexigenic compensatory mechanisms are operative, aimed at maintaining a "normal" feeding pattern. Thus, the occurrence of a primary, age-related alteration in the feeding mechanisms is unlikely.

In order to verify the effects of the sporting season (entailing periods of training, competition, recovery, resting) on GH-dependent parameters in male and female athletes from different sporting disciplines, 47 male and female athletes (3 rowers, 5 swimmers, 7 alpine skiers, 3 soccer players, 7 middle distance runners, 14 sprinters, 4 triathletes, 1 road walker, 3 cyclists) were followed-up for a period of 6 months. Blood samples were taken every two months for the evaluation of IGF-I, N-terminal propeptide of type III procollagen (PIIINP) and C-terminal cross-linked telopeptide of type I collagen (ICTP). Abnormal IGF-I, PIIINP and ICTP levels were observed during the follow-up period in 7/100 (7%), 9/100 (9.0%) and 8/100 (8%) samples of the male group, respectively, and in 9/88 (10.2%), 1/88 (1.1%) and 0/88 (0%) samples of the female group, respectively. Abnormal levels appeared to be randomly distributed over the different periods of the sporting season and within male and female subjects, with the large majority of abnormal values being found in the younger athletes. Taking into account all the tests done during the 6-month period (no. 564), individual markers falling outside the normal range (for age) were observed in a small number of instances (34/564 tests done, 24/300 for males and 10/264 for females). When our method for the detection of exogenous recombinant GH (rhGH) administration, based on the concomitant determination of these three peripheral GH-dependent markers and on the attribution of specific scores, was applied in the same athlete at a given time point of the 6-month period, the prevalence of a positive score was extremely low (ie, 3/188 samples or 1.6%). Total positive scores were actually recorded in only three male athletes (2 swimmers and 1 skier, aged <21 yr) at one occasion during the 6-month period considered. In contrast, no total positive scores were found in female athletes (ie, 0/88 samples). In conclusion, the concentrations of IGF-I, PIIINP and ICTP were stable and not significantly modified during 6 months of a sporting season (entailing periods of training, competition, recovery, resting) in athletes from different sporting disciplines. Therefore our method, based on the concomitant determination of three peripheral GH-dependent biomarkers appears safe, acceptable, relatively inexpensive and repeatable (in case of positive or suspected values) immediately or at different intervals of the sporting season. Further additional studies are requested to precise the cut-off values for narrower age-class subdivisions in both genders in order to improve the proposed method.