Growth and survival of the native oyster Crassostrea gasar cultured under different stocking densities in two grow-out systems in tropical climate / [Crescimento e sobrevivência da ostra nativa Crassostrea gasar cultivada em diferentes densidades em dois sistemas em clima tropical]

Survival and growth of the native oyster Crassostrea gasar along the juvenile and adult phases were evaluated in three different stocking densities [low (D), medium (2D) and high (3D)] and in two grow-out systems (fixed and floating system). The fixed system consisted of a rack made with PVC, fixed from the bottom with wood sticks. The floating system consisted of floating bags suspended by a rack made with PVC and maintained submerged from the seawater surface by eight floats. Survival and shell height of oysters cultured after 30, 60 and 90 days were registered in each phase and in each grow-out system. Results showed that the grow-out system did not affect survival and growth of C. gasar in the juvenile and adult phases. The tested densities affected the survival of oysters cultured over time in both phases but did not affect oyster growth. At times analyzed, it was observed positive growth in juvenile oysters grow after 90 days of culture. However, in the adult phase, no growth was observed after 90 days of culture. Oyster yield was higher in the density 3D, in both juvenile and adult phases. These findings contributed to the development of the oyster C. gasar culture.(AU)

A sobrevivência e o crescimento da ostra nativa Crassostrea gasar nas fases juvenil e adulta foram avaliados sob três diferentes densidades de estocagem [baixa (D), média (2D) e alta (3D)] e dois sistemas de engorda (fixo e flutuante). O sistema fixo consistiu em uma mesa de PVC, fixada na parte inferior com varas de madeira. O sistema flutuante consistiu em travesseiros flutuantes suspensos por uma mesa de PVC e mantidas submersas da superfície da água do mar por oito flutuadores. Registraram-se sobrevivência e altura da concha de ostras cultivadas após 30, 60 e 90 dias, em cada fase (juvenil e adulta) e em cada sistema (fixo e flutuante). Os resultados mostraram que o sistema de engorda não afetou a sobrevivência e o crescimento de C. gasar nas fases juvenil e adulta. As densidades testadas afetaram a sobrevivência das ostras ao longo do tempo, em ambas as fases, mas não afetaram o crescimento em altura. Nos tempos analisados, ostras juvenis apresentaram crescimento após 90 dias de cultivo. Porém, na fase adulta, não foi observado crescimento após 90 dias de cultivo. A produção de ostras, foi maior na densidade 3D, nas fases juvenil e adulta. Os presentes achados contribuíram para o desenvolvimento do cultivo da ostra C. gasar.(AU)

Clone worth? Genetic diversity in obligate apomictic Miconia albicans (Melastomataceae).

Apomixis is the asexual production of seeds by plants and, in theory, would render low genetic diversity and even clonal lineages. However, recent studies have shown otherwise, although is not always clear where the genetic diversity of obligate apomicts comes from. We evaluated the genetic diversity among sister seedlings of M. albicans, an obligate apomictic species in Cerrado, Neotropical Savannas in Central Brazil. A total of 50 seedlings from five individuals were analysed using ISSR primers. We obtained 107 fragments, all with good resolution, consistently observed and replicable. The percentage of polymorphic loci ranged from 28.04% to 33.64% and Shannon's information index (I) averaged 0.173. The expected heterozygosity (He) averaged 0.117, similar to the observed for populations of M. albicans and other selfed species. Only two seedlings showed the same genotype (possible clones), but most differed at least for five loci. Most of variance was among progenies (62%), but we found that 38% was within progenies. Genetic distances separated the progenies in two groups, and analogous analyses between individuals reconstructed the original progenies clustering. The results confirmed a relatively high genetic diversity among sister seedling of this obligatory apomictic plant and clones were rare. This diversity can be generated during development, probably by restitutional meiosis or other recombination processes. These differences may accumulate into lineages and populations well adapted to heterogenous Cerrado environment.

The single recombinant M. tuberculosis protein DPPD provides enhanced performance of skin testing among HIV-infected tuberculosis patients.

Diagnostic testing for M. tuberculosis infection has advanced with QuantiFERON and GeneXpert, but simple cost-effective alternatives for widespread TB screening has remained elusive and purified protein derivative (PPD)-based tuberculin skin testing (TST) remains the most widely used method. PPD-based tests have reduced performance, however, in BCG vaccinees and in individuals with immune deficiencies. We compared the performance of skin testing with the recombinant DPPD protein against that of a standard PPD-based skin test. Our data indicates similar performance of DPPD and PPD (r2 = 0.7689) among HIV-negative, active TB patients, all of whom presented greater than 10 mm induration following administration. In contrast to results demonstrating that PPD induced indurations greater than 5 mm (i.e., the recommended threshold for positive results in this population) in only half (19 of 38) of the HIV positive TB patients, 89.5% (34 of 38) of these participants developed indurations greater than 5 mm when challenged with DPPD. Importantly, none of the patients that were positive following PPD administration were negative following DPPD administration, indicating markedly improved sensitivity of DPPD among HIV-infected individuals. Our data indicate that DPPD has superior performance in skin testing than the current TST standard.

Unexpectedly high genetic diversity and divergence among populations of the apomictic Neotropical tree Miconia albicans.

Since tropical trees often have long generation times and relatively small reproductive populations, breeding systems and genetic variation are important for population viability and have consequences for conservation. Miconia albicans is an obligate, diplosporous, apomictic species widespread in the Brazilian Cerrado, the savanna areas in central Brazil and elsewhere in the Neotropics. The genetic variability would be, theoretically, low within these male-sterile and possibly clonal populations, although some variation would be expected due to recombination during restitutional meiosis. We used ISSR markers to assess genetic diversity of M. albicans and to compare with other tropical trees, including invasive species of Melastomataceae. A total of 120 individuals from six populations were analysed using ten ISSR primers, which produced 153 fully reproducible fragments. The populations of M. albicans presented mean Shannon's information index (I) of 0.244 and expected heterozygosity (He ) of 0.168. Only two pairs of apparently clonal trees were identified, and genetic diversity was relatively high. A hierarchical amova for all ISSR datasets showed that 74% of the variance was found among populations, while only 26% of the variance was found within populations of this species. Multivariate and Bayesian analyses indicated marked separation between the studied populations. The genetic diversity generated by restitutional meiosis, polyploidy and possibly other genome changes may explain the morpho-physiological plasticity and the ability of these plants to differentiate and occupy such a wide territory and different environmental conditions. Producing enormous amounts of bird-dispersed fruits, M. albicans possess weedy potential that may rival other Melastomataceae alien invaders.

Mixed pollen load and late-acting self-incompatibility flexibility in Adenocalymma peregrinum (Miers) L.G. Lohmann (Bignonieae: Bignoniaceae).

Mixed cross and self-pollen load on the stigma (mixed pollination) of species with late-acting self-incompatibility system (LSI) can lead to self-fertilized seed production. This "cryptic self-fertility" may allow selfed seedling development in species otherwise largely self-sterile. Our aims were to check if mixed pollinations would lead to fruit set in LSI Adenocalymma peregrinum, and test for evidence of early-acting inbreeding depression in putative selfed seeds from mixed pollinations. Experimental pollinations were carried out in a natural population. Fruit and seed set from self-, cross and mixed pollinations were analysed. Further germination tests were carried out for the seeds obtained from treatments. Our results confirm self-incompatibility, and fruit set from cross-pollinations was three-fold that from mixed pollinations. This low fruit set in mixed pollinations is most likely due to a greater number of self- than cross-fertilized ovules, which promotes LSI action and pistil abortion. Likewise, higher percentage of empty seeds in surviving fruits from mixed pollinations compared with cross-pollinations is probably due to ovule discounting caused by self-fertilization. Moreover, germinability of seeds with developed embryos was lower in fruits from mixed than from cross-pollinations, and the non-viable seeds from mixed pollinations showed one-third of the mass of those from cross-pollinations. The great number of empty seeds, lower germinability, lower mass of non-viable seeds, and higher variation in seed mass distribution in mixed pollinations, strongly suggests early-acing inbreeding depression in putative selfed seeds. In this sense, LSI and inbreeding depression acting together probably constrain self-fertilized seedling establishment in A. peregrinum.

Vocal function and vocal discomfort in sheltered and non-sheltered 7- to 10-year-old school children in Aracaju, Brazil.

OBJECTIVE: To compare vocal function, school performance, and vocal discomfort between sheltered and nonsheltered school children in Aracaju, Brazil. METHODS: A controlled cross-sectional study was carried out on 7- to 10-year-old children who attended school regularly. Two groups of children were studied: the study group (SG), with children who lived in a shelter, and the control group (CG) containing children who lived with their families. We interviewed 44 children for the SG and 15 (34%) revealed vocal discomfort (SG = 15). Concomitantly, we interviewed 400 regular school children from the same geographical area and 45 (11.25%) were selected for the control group (CG). They were paired by sex and age with the sheltered children using a 3:1 ratio. Both groups were interviewed about school performance and vocal discomfort and were evaluated using perceptual and acoustic measurements for the voice and larynx. RESULTS: Children from both groups had started public school late. There were more individuals with vocal discomfort in the SG and individuals in this group also had a slower speech rate and inadequate pneumophonic coordination compared with the CG. The Dysphonia Severity Index (DSI) revealed mild-moderate deviation for both groups. Upper harmonics and palatal tonsil hypertrophy were higher in the CG, whereas laryngeal constriction was more common in the SG. CONCLUSION: All the SG children revealed mild-moderate deviance on the DSI, a higher level of vocal discomfort, a slow speech rate, inadequate pneumophonic coordination, and laryngeal constriction. The results here presented suggest that social conditions are important for voice behavior in children.

Voice performance evaluation of street children from Aracaju, Brazil using perceptual-auditory and acoustic analyses.

OBJECTIVE: To compare the voice performance of children involved in street labor with regular children using perceptual-auditory and acoustic analyses. METHODS: A controlled cross-sectional study was carried out on 7- to 10-year-old children of both genders. Children from both groups lived with their families and attended school regularly; however, child labor was evident in one group and not the other. A total of 200 potentially eligible street children, assisted by the Child Labor Elimination Programme (PETI), and 400 regular children were interviewed. Those with any vocal discomfort (106, 53% and 90, 22.5%) had their voices assessed for resonance, pitch, loudness, speech rate, maximum phonation time, and other acoustic measurements. RESULTS: A total of 106 street children (study group [SG]) and 90 regular children (control group [CG]) were evaluated. The SG group demonstrated higher oral and nasal resonance, reduced loudness, a lower pitch, and a slower speech rate than the CG. The maximum phonation time, fundamental frequency, and upper harmonics were higher in the SG than the CG. Jitter and shimmer were higher in the CG than the SG. CONCLUSION: Using perceptual-auditory and acoustic analyses, we determined that there were differences in voice performance between the two groups, with street children having better quality perceptual and acoustic vocal parameters than regular children. We believe that this is due to the procedures and activities performed by the Child Labor Elimination Program (PETI), which helps children to cope with their living conditions.

Relationship between social capital and test seeking among blood donors in Brazil.

BACKGROUND AND OBJECTIVES: Higher risk of HIV infection could be associated with test seeking, which is one motivation for donating blood. Cognitive social capital is defined as the social support, trust and co-operation that guide community behaviour. Structural social capital refers to an individual's participation in institutions and organizations. The association between social capital and test seeking was assessed. MATERIALS AND METHODS: A survey of over 7500 donors in three Brazilian blood centres was conducted. Test seeking was classified into four non-overlapping categories (non-test seeker, possible, presumed and self-disclosed test seekers) using one direct and two indirect questions. Social capital was summarized into cognitive and structural categorizations. Multivariable logistic regression analysis was performed. RESULTS: Compared with non-test seekers (62% of survey respondents), cognitive social capital was higher for each category of test seeking (OR=1.1, 7.4, 7.1, P<0.05 respectively). Male gender, lower education and lower income were also significantly associated with test seeking. CONCLUSION: As test seekers appear to have strong social networks, blood banks may leverage this to convince them to seek testing at other locations.

Mating in the pseudogamic apomictic Anemopaegma acutifolium DC: another case of pseudo-self-compatibility in Bignoniaceae?

Self-compatibility in apomictic pseudogamic species is considered fundamental to assure reproduction by seeds in extreme situations, making apomictic species more advantageous than sexual ones in these scenarios. Anemopaegma acutifolium is a polyploidy, apomictic sporophytic species with no endosperm development in ovules of unpollinated pistils, which indicates obligate pseudogamy. Thus, the aim of the present work is to study the breeding system and post-pollination events to test if there is similar pseudogamous development irrespective of pollination treatment. We analysed fruit and seed set obtained in controlled experimental pollinations, as well as embryo number per seed, and the progress of ovule penetration, fertilisation and early endosperm development between self- and cross-pollinated pistils. We found that the species is self-fertile and that spontaneous selfing fruit set is also possible, although emasculated flowers never form fruits. Selfed pistils were as efficient as crossed ones for all parameters analysed, except for a delay in endosperm development observed in the former that may be an effect of the late-acting self-incompatibility. Therefore, the avoidance of selfed pistil abortion seems to be promoted by the presence of adventitious embryos and a normal endosperm. We conclude that A. acutifolium shows apomixis-related pseudo-self-compatibility, as in other self-fertile apomictic species of Bignoniaceae, which confer reproductive assurance and increases fruit-set and persistence ability in fast-changing tropical habitats.

High prevalence of micropenis in 2710 male newborns from an intensive-use pesticide area of Northeastern Brazil.

Exposure to endocrine-disrupting chemicals (EDCs) has been suggested to contribute to the increasing trends of external genital malformation in male newborns. In Northeastern Brazil, the poor sanitary conditions found in the favelas encourage the widespread use of pesticides. This 2-year study of a total birth cohort of full-term male newborns in the regional hospitals of Campina Grande (Paraíba, Brazil) sought to (1) accurately establish for the first time the incidences of neonatal male genital malformations, (2) investigate the endocrine and genetic aetiologies of these malformations, and (3) evaluate their associations with possible prenatal exposure to EDCs. A total of 2710 male newborns were explored for cryptorchidism, hypospadias and micropenis. Cases were referred to the Pediatric Endocrine Clinic for endocrine and genetic investigations, and all parents were interviewed about their environmental/occupational exposure to EDCs before/during pregnancy by paediatric endocrinologists using a detailed questionnaire. We observed 56 cases of genital malformation (2.07%), including 23 cryptorchidism (0.85%), 15 hypospadias (0.55%), and 18 micropenis (0.66%). All cases exhibited normal/subnormal testosterone production and none presented androgen receptor or 5α-reductase gene mutation. More than 92% of these newborns presented foetal contamination by EDCs, as their mothers reported daily domestic use of pesticides (i.e., DDT) and other EDCs. Most of these undervirilized male newborns presented additional EDC contamination, as 80.36% of the mothers and 58.63% of the fathers reported paid or unpaid work that entailed the use of pesticides and other EDCs before/during pregnancy for the mothers and around the time of fertilization for the fathers. The high rate of micropenis in our population associated with an elevated percentage of parental environmental/occupational exposure to EDCs before/during pregnancy indicates that foetal contamination may be a risk factor for the development of male external genital malformation.

Obstrução intestinal por Pythium insidiosum em um cão: relato de caso / Intestinal obstruction by Pythium insidiosum in a dog: case report

Relata-se um caso de pitiose intestinal em um Husky Siberiano, de um ano de idade, macho atendido em um Hospital Veterinário Escola com sinais de obstrução intestinal. Ao exame físico, notou-se massa cilíndrica na região abdominal cranial, posteriormente confirmada por meio de radiografia e ultrassonografia. Durante a laparotomia exploratória, constatou-se massa extraluminal envolvendo o jejuno e alterações na parede do órgão. Realizou-se a ressecção da porção afetada do intestino e, posteriormente, anastomose. O exame histológico do tecido evidenciou inflamação piogranulomatosa acentuada. Na coloração de prata metenamina de Grocott, hifas septadas foram observadas. O diagnóstico de infecção por Pythium insidiosum foi confirmado por meio da imunoistoquímica. Após a cirurgia, o animal restabeleceu a defecação; no 30º dia pós-cirúrgico, foi relatada ainda presença de diarreia. O tratamento com itraconazol e terbinafina foi instituído durante 60 dias. Após dois anos do procedimento cirúrgico e do tratamento com antifúngicos orais, o cão não apresentou recidiva.

A case of intestinal pythiosis in a one-year-old male Siberian Husky treated at a Veterinary School Hospital with signs of intestinal obstruction is reported. At physical examination, a cylindrical mass was palpable in the cranial abdomen, later confirmed by radiography and ultrasonography. During the exploratory laparotomy, it was evidenced an extraluminal mass involving the jejunum and alterations of the wall in the organ. After that, a resection of the affected portion of the intestine was made followed by anastomosis. The histologic examination evidenced accented pyogranulomatous inflammation. By the Grocott methenamine silver stain, branching hyphae were observed. The diagnosis of infection by Pythium insidiosum was confirmed by immunohistochemistry. The animal reestablished the defecation after the surgery; on the 30th postoperative day, it was also reported the presence of diarrhea. The treatment with itraconazol and terbinafina was instituted for 60 days. Two years after the surgical procedure and treatment with oral antifungals, the dog did not show recurrence.

Antinociceptive properties in mice of a lectin isolated from the marine alga Amansia multifida Lamouroux.

The antinociceptive effects of a lectin (LEC) isolated from the marine alga Amansia multifida were determined in Swiss mice. The LEC (1, 5, and 10 mg/kg) inhibited acetic acid-induced abdominal writhings in a dose-dependent manner after intraperitoneal or oral administration. A partial but significant inhibition of writhings was observed after the combination of LEC (10 mg/kg) with avidin (1 mg/kg), a potent inhibitor of the hemmaglutinant activity of the lectin. However, total writhing inhibition was demonstrable in the group of mice treated with LEC plus mannose (1 mg/kg), as compared to LEC alone or to control groups. Furthermore, avidin and mainly mannose also play a role in antinociception, somehow facilitating the interaction of LEC with its active cell sites. In the formalin test, although both phases of the response were significantly inhibited, the effect of LEC was predominant during phase 2, causing inhibition of licking time that ranged from 48 to 88% after oral (5 and 10 mg/kg) and intraperitoneal (1 to 5 mg/kg) administration. As is the case with morphine, the effect of LEC (2 mg/kg) was reversed by naloxone (2 mg/kg), indicating the involvement of the opioid system. LEC was also effective in the hot-plate test, producing inhibitory responses to the thermal stimulus, and its effects were blocked by naloxone. In the pentobarbital-induced sleeping time, although LEC did not alter the onset of sleep significantly, it increased the time of sleep within the same dose range compared to control. These results show that LEC presents antinociceptive effects of both central and peripheral origin, possibly involving the participation of the opioid system.

Antinociceptive properties in mice of a lectin isolated from the marine alga Amansia multifida Lamouroux

The antinociceptive effects of a lectin (LEC) isolated from the marine alga Amansia multifida were determined in Swiss mice. The LEC (1, 5, and 10 mg/kg) inhibited acetic acid-induced abdominal writhings in a dose-dependent manner after intraperitoneal or oral administration. A partial but significant inhibition of writhings was observed after the combination of LEC (10 mg/kg) with avidin (1 mg/kg), a potent inhibitor of the hemmaglutinant activity of the lectin. However, total writhing inhibition was demonstrable in the group of mice treated with LEC plus mannose (1 mg/kg), as compared to LEC alone or to control groups. Furthermore, avidin and mainly mannose also play a role in antinociception, somehow facilitating the interaction of LEC with its active cell sites. In the formalin test, although both phases of the response were significantly inhibited, the effect of LEC was predominant during phase 2, causing inhibition of licking time that ranged from 48 to 88 percent after oral (5 and 10 mg/kg) and intraperitoneal (1 to 5 mg/kg) administration. As is the case with morphine, the effect of LEC (2 mg/kg) was reversed by naloxone (2 mg/kg), indicating the involvement of the opioid system. LEC was also effective in the hot-plate test, producing inhibitory responses to the thermal stimulus, and its effects were blocked by naloxone. In the pentobarbital-induced sleeping time, although LEC did not alter the onset of sleep significantly, it increased the time of sleep within the same dose range compared to control. These results show that LEC presents antinociceptive effects of both central and peripheral origin, possibly involving the participation of the opioid system.

Cognitive restraint is associated with higher intake of vegetables in a sample of university students.

The main objective of this study was to determine the association between cognitive restraint, nutritional intake and eating patterns in free-leaving subjects. We administered a questionnaire that included information about eating behaviour (restraint, disinhibition, and hunger), dietary intake, and physical activity; 380 students (60% female) completed the study. The association of restraint (low/high) and disinhibition (low/high) with dietary intake was tested using MANOVA in a 2x2 factorial design. Statistically significant main effects were further analyzed using ANOVAs. To identify eating patterns, factorial analysis was employed. Among women, high restrainers reported lower consumption of energy, pastry, and starchy foods, and higher consumption of vegetables, and fish, than low restrainers. In male subjects, high restrainers consumed significantly more vegetables than low restrainers. The major food pattern in female restrainers aggregate: higher consumption of legumes and fruit intake; and lower consumption of pastry, sugar, and starchy foods. In restrained men, the more important food pattern consists of vegetable soup, fruit, milk, and eggs. These differences should be considered in clinical interventions for individuals seeking a better body weight control.

Co-infection with HTLV-1 is associated with a shorter survival time for HIV-1-infected patients in Bahia, Brazil.

Co-infection with HTLV-1 reaches 20% among patients infected by HIV-1 in Bahia, Brazil. To evaluate its impact on survival, we conducted a retrospective, case-control study involving 198 patients (63 cases). Co-infection was associated with parenteral exposure (P = 0.0001) and female sex (P = 0.02). Co-infected patients had a shorter mean survival (1849 days) than controls (2430 days, P = 0.001), regardless of sex or baseline CD4 cell count. In Bahia, Brazil, co-infection with HIV-1 and HTLV-1 is associated with a shorter survival time.

[Epidemiology of anorexia nervosa. Prevalence of anorexia nervosa in female adolescents from the districts of Lisboa and Setubal]. / Epidemiologia da anorexia nervosa. Prevalência da anorexia nervosa em adolescentes do sexo feminino nos distritos de Lisboa e Setúbal.

UNLABELLED: Prevalence studies of anorexia nervosa (AN) have shown differences depending of the country and method applied. The partial syndrome is being emphasized. There has been no epidemiological study done on AN in the Portuguese population of continental Portugal. Our objectives were: 1) to study the prevalence of AN in Portuguese populations; 2) to study the presence of partial syndrome and of body image disturbance; 3) to calculate the number of girls dieting; 4) to correlate with other variables. A population of 2,398 girls ranging 10 to 21 years old, from 30 State secondary schools in the Lisbon and Setubal districts were studied. A questionnaire was answered with questions allowing assessment of all DSM IIIR criteria for AN. RESULTS: Prevalence of AN--0.37%, partial syndrome--12.6%, body image disturbance without weight loss--7%, wish to decrease weight in normal or low weight girls--38%, overweight--15.3%, the peak of age for AN was 15 years, the most prevalent socio-economic class for AN was 2 (scale 1 (higher) to 5). School performance was worse with AN and the age menarche was sooner.

Serological expression cloning and immunological evaluation of MTB48, a novel Mycobacterium tuberculosis antigen.

Improved diagnostics are needed for the detection of Mycobacterium tuberculosis, especially for patients with smear-negative disease. To address this problem, we have screened M. tuberculosis (H37Rv and Erdman strains) genomic expression libraries with pooled sera from patients with extrapulmonary disease and with sera from patients with elevated reactivity with M. tuberculosis lysate. Both serum pools were reactive with clones expressing a recombinant protein referred to here as MTB48. The genomic sequence of the resulting clones was identical to that of the M. tuberculosis H37Rv isolate and showed 99% identity to the Mycobacterium bovis and M. bovis BCG isolate sequences. The genomic location of this sequence is 826 bp upstream of a region containing the esat-6 gene that is deleted in the M. bovis BCG isolate. The mtb48 1,380-bp open reading frame encodes a predicted 47.6-kDa polypeptide with no known function. Southern and Western blot analyses indicate that this sequence is present in a single copy and is conserved in the M. tuberculosis and M. bovis isolates tested but not in other mycobacterial species tested, including Mycobacterium leprae and Mycobacterium avium. In addition, the native protein was detected in the cytoplasm, as was a processed form that was also shed into the medium during culture. Serological analysis of recombinant MTB48 and the M. tuberculosis 38-kDa antigen with a panel of patient and control sera indicates that the inclusion of recombinant MTB48 in a prototype serodiagnostic test increases assay sensitivity for M. tuberculosis infection when it is combined with other known immunodominant antigens, such as the 38-kDa antigen.

Assessment of the serodiagnostic potential of nine novel proteins from Mycobacterium tuberculosis.

To identify antigens that would improve the accuracy of serological diagnosis of active tuberculosis, we cloned the genes encoding nine potentially immunogenic secreted or surface-associated proteins of Mycobacterium tuberculosis. Recombinant proteins were reacted with sera from HIV-negative individuals with extrapulmonary tuberculosis (EP-TB) or HIV-positive individuals with pulmonary tuberculosis (TBH). Specific and high level antibody responses were obtained for four recombinant proteins, of which antigen GST-822 was recognized by 60% of EP-TB and 42% of TBH and antigen MBP-506 was recognized by 45% of EP-TB and 61% of TBH. These results suggest that these proteins are strong candidates as subunits in a polyvalent serodiagnostic test.

Evaluation of DPPD, a single recombinant Mycobacterium tuberculosis protein as an alternative antigen for the Mantoux test.

Although the tuberculin test has aided in the diagnosis of tuberculosis for more than 85 years, its interpretation is difficult particularly because sensitization with non-tuberculous mycobacteria leads to false positive tests. Using the guinea pig model of tuberculosis, we have recently described a recombinant antigen (DPPD) that could circumvent this problem. The DPPD gene is unique to the M. tuberculosis complex organisms and is absent in the organisms representative of all other members of the Mycobacterium genus. Moreover, DPPD induced strong DTH in 100% of the guinea pigs infected with M. tuberculosis and in none of the guinea pigs immunized with nine different species of Mycobacterium. Here we present results of a clinical investigation using DPPD. Mantoux test using both PPD and DPPD was initially performed in 26 patients with confirmed pulmonary tuberculosis and in 25 healthy PPD negative individuals. The results indicated that both PPD and DPPD elicited DTH in 24 out of the 26 patients. No DTH was observed in any of the PPD negative individuals. In addition, a small clinical trial was performed in a population of 270 clinically healthy and randomly selected individuals. DPPD produced a bimodal histogram of skin reaction size and PPD produced a skewed histogram. Because the DPPD gene is not present in non-tuberculous bacilli, these results suggest that this molecule can be an additional tool for a more specific diagnosis of tuberculosis.

High prevalence of giardiasis and stronglyloidiasis among HIV-infected patients in Bahia, Brazil.

Diarrhea due to intestinal microbial infections is a frequent manifestation among HIV-infected patients. It has been postulated that HIV-infected patients may have special types of intestinal infections, and that immune activation from such parasites may affect the progression of HIV disease. To evaluate these associations, the frequency of infections was examined in HIV-infected patients in Bahia, Brazil. To determine the potential impact of the presence of intestinal parasitic infections on HIV disease progression, a retrospective study approach was used. The medical charts of 365 HIV-infected patients who had been treated at the AIDS Clinic of the Federal University of Bahia Hospital were reviewed, and the prevalence of parasites was compared with 5,243 HIV-negative patients who had attended the hospital during the same period of time. Among HIV-infected subjects, CD(4) count, RNA plasma viral load (VL), and number of eosinophils were compared according to their stool examination results. The overall prevalence of each parasite was similar for HIV-positive and HIV-negative patients. However, the prevalence of S. stercoralis (p<10(-7)) and G. lamblia (p=0.005) was greater for HIV-infected subjects. The mean CD(4) count and viral load of HIV patients in our clinic who had stool examinations was 350 cells +/- 340 and 4.4 +/- 1.4 log RNA viral load, respectively. In this patient group there was no clear association between the level of the absolute CD(4) count or the viral load and a specific parasitic infection. The presence of an intestinal parasitic infection was not associated with faster progression of the HIV disease among HIV-infected patients. We conclude that strongyloidiasis and giardiasis are more frequent in HIV-infected patients in Bahia, Brazil. If this association is due to immune dysregulation, as has been proposed elsewhere, it must occur in patients after only minor shifts in CD(4) count from normal levels, or as a result of immune dysfunction not represented by CD(4) count. These infections do not appear to alter the progression of HIV disease.