Effect of mineral status and glucocorticoid use on bone mineral density in patients with Crohn's disease.

OBJECTIVES: Crohn's disease (CD) is a condition that is characterized by chronic inflammation. The presence of multifactorial pathogenesis that results from inflammation is associated with low micronutrient consumption and glucocorticoid use, which may be related to bone health. This study aimed to evaluate the relationship between dietary mineral intake and glucocorticoid use in bone mineral density (BMD) in patients with CD. METHODS: A cross-sectional study of 62 patients with CD ages 20 y to 40 y measured their macro- and micronutrient intake with a 3-d food record. The lumbar spine and femoral neck BMDs were determined using a bone densitometry technique. The C-reactive protein (CRP) levels and erythrocyte sedimentation rate (ESR) values were also noted. RESULTS: Dietary intake of calcium, zinc, and magnesium was below the reference values but the phosphorus intake level was within the normal value range. Patients with osteopenia and osteoporosis accounted for 17.7% and 14.5%, respectively, of the total number of participants. Significant bone loss was found in 22.6% of patients taking glucocorticoid medications. BMD was significantly reduced and also observed in patients in the active phase of their disease. Zinc and calcium intakes were found to be correlated with reduced femoral neck BMD. The mean CRP and ESR values were above the normal ranges. Significant differences in age and ESR were observed between patients with normal and reduced BMD (P <0.05). CONCLUSIONS: Low calcium and zinc intake, glucocorticoid use, and active disease phase are favorable conditions for bone loss in patients with Crohn's disease.

The anterior sagittal transrectal approach (ASTRA) for cases associated with rectal implantation of the urethra: A retrospective review of six cases.

INTRODUCTION: Severe genital abnormalities such as urogenital sinus and urethral duplication with ectopic urethra in the rectum represent a major challenge in reconstructive urology. OBJECTIVE: We aimed to review our cases presenting with functional ectopic urethra implanted in the rectum that were treated through an ASTRA approach. METHODS: We reviewed the medical records of all patients who had undergone an ASTRA approach from 2005-2016. We collected data with interest to primary diagnosis, clinical presentation, additional procedure, immediate clinical outcome, complications, bowel habits after surgery, voiding and bladder emptying pattern and long term follow-up. RESULTS: Since 2005, we treated 6 cases using this method, consisting of 3 patients with congenital aphallia and 3 with Y-type urethral duplication. Two aphallia patients underwent De Castro's neophalloplasty with simultaneous anastomosis of proximal urethra to a tubed buccal mucosa neourethra and one had a neophalloplasty with transverse skin flaps and primary perineal urethrostomy. Two patients with Y-type urethral duplication had a complete urethroplasty performed (one end-to-end anastomosis and a two-stage repair). Last patient had a definitive perineal urethrostomy. At mean follow-up of 83.5 meses, only one patient voids through the urethra, four have a Mitrofanoff channel and two have a perineal urethrostomy. Immediate follow-up was uneventful and none of our patients had any bowel complications nor fecal incontinence. DISCUSSION: Domini et al. were the first proponents of the anterior sagittal transanorectal approach (ASTRA) as an alternative to classic Peña approach. Later, De Castro popularized specifically this technique as a relevant part of his neophalloplasty procedure to treat congenital aphallia. We did not find in this series any complications related to ASTRA technique in regards to bowel habits, fecal incontinence or infection but most of cases we tried to create a new anterior urethra and connect to the proximal stump failed. Moreover, we are skeptic to support simultaneous urethral repair when there is a dysplastic bulbar segment between the proximal stump and the end of the penile urethra normally at penoscrotal junction area by urethral duplication. Treatment at two-stages is also prone to fail in the midterm. At end, most of them will have either a Mitrofanoff or perineal urethral stoma as shown in our data. CONCLUSION: The ASTRA approach proved to be useful for cases of rectal implantation of the urethra. No complications related to anterior sphincter incision were documented. We believe it should be regarded as the treatment of choice for cases alike ours.

FISH and AgNor mapping of the 45S and 5S rRNA genes in wild and cultivated species of Capsicum (Solananceae).

Chromosome number and position of rDNA were studied in 12 wild and cultivated species of the genus Capsicum with chromosome numbers x = 12 and x = 13 (22 samples). For the first time in these species, the 5S and 45S rRNA loci were localized and physically mapped using two-color fluorescence in situ hybridization and AgNOR banding. We focused on the comparison of the results obtained with both methods with the aim of accurately revealing the real functional rRNA genes. The analyzes were based on a previous work that reported that the 18S-5.8S-25S loci mostly coincide with GC-rich heterochromatic regions and likely have given rise to satellite DNAs, which are not active genes. These data show the variability of rDNA within karyotypes of the genus Capsicum, providing anchor points for (comparative) genetic maps. In addition, the obtained information might be useful for studies on evolution of repetitive DNA.

Autoimmunity, phospholipid-reacting antibodies and malaria immunity.

Several questions regarding the production and functioning of autoantibodies (AAb) during malaria infection remain open. Here we provide an overview of studies conducted in our laboratory that shed some light on the questions of whether antiphospholipid antibodies (aPL) and other AAb associated with autoimmune diseases (AID) can recognize Plasmodia antigens and exert anti-parasite activity; and whether anti-parasite phospholipid antibodies, produced in response to malaria, can inhibit phospholipid-induced inflammatory responses and protect against the pathogenesis of severe malaria. Our work showed that sera from patients with AID containing AAb against dsDNA, ssDNA, nuclear antigens (ANA), actin, cardiolipin (aCL) and erythrocyte membrane antigens recognize plasmodial antigens and can, similarly to monoclonal AAb of several specificities including phospholipid, inhibit the growth of P. falciparum in vitro. However, we did not detect a relationship between the presence of anti-glycosylphosphatidylinositol (GPI) antibodies in the serum and asymptomatic malaria infection, although we did register a relationship between these antibodies and parasitemia levels in infected individuals. Taken together, these results indicate that autoimmune responses mediated by AAb of different specificities, including phospholipid, may have anti-plasmodial activity and protect against malaria, although it is not clear whether anti-parasite phospholipid antibodies can mediate the same effect. The potential effect of anti-parasite phospholipid antibodies in malarious patients that are prone to the development of systemic lupus erythematosus or antiphospholipid syndrome, as well as the (possibly protective?) role of the (pathogenic) aPL on the malaria symptomatology and severity in these individuals, remain open questions.

Fitoterápicos: uma abordagem farmacotécnica / Herbal medicines: Pharmacotechnique approach

O artigo, de cunho informativo e crítico, propõe uma perspectiva de desenvolvimento de fitoterápicos sob a ótica da interdisciplinaridade, que envolve as inúmeras especificidades das diversas áreas que compõem o referido tema. Enumera-se a pesquisa de acordo com a antropologia botânica, a botânica, a agronomia, a fitoquímica, a farmacologia, a toxicologia, os aspectos de desenvolvimento e de estudo clínico do fitoterápico. Nesta perspectiva discute-se o fitoterápico inserido em uma proposta de construção de monografias nacionais de modo a contribuir com a indústria, com os centros estaduais, municipais de produção e distribuição de medicamentos, proporcionando dentre as opções de terapêutica um medicamento ético, seguro e eficiente.

Plasmodium falciparum: limited genetic diversity of MSP-2 in isolates circulating in Brazilian endemic areas.

The genetic polymorphism of the surface merozoite protein 2 (MSP-2) was evaluated in Plasmodium falciparum isolates from individuals with uncomplicated malaria living in a Brazilian endemic area of Peixoto de Azevedo. The frequency of MSP-2 alleles and the survival of genetically different populations clones in 104 isolates were verified by Southern blot and SSCP-PCR. Single and mixed infections were observed in similar frequencies and the rate of detection of FC27 and 3D7 allelic families was equivalent. Eight alleles were identified and among them, the sequence polymorphism was mainly attributed to variations in the repetitive region. Interestingly, in three alleles nucleotide polymorphism was identical to that detected in a previous study, conducted in 1992, in a near Brazilian endemic area. This finding demonstrated the genetic similarity between two isolate groups, besides the certain temporal stability in the allelic patterns. The implications of these data for studies on the genetic diversity are also discussed.

Matrilineal genetic structure within and among populations of the cooperatively breeding common marmoset, Callithrix jacchus.

Common marmosets are members of the family Callitrichidae, South American primates characterized by highly social group living and cooperative breeding. In this study we analysed 1112 base pairs (bp) of the mitochondrial control region in 59 Callithrix jacchus individuals, sampled mainly from two geographically distinct field sites in N.E. Brazil. Analysis of molecular variation revealed a highly significant genetic structuring of haplotypes between social groups and between populations. Examination of matrilineal genetic structure within social groups revealed that seven of nine recorded breeding pairs were from different maternal lineages, indicating assortative mating and outbreeding. In addition to the breeders, at least six of 10 groups contained adult individuals from different matrilines, with five haplotypes present in one social group of nine animals. Groups of mixed lineages raise questions about potential reproductive conflicts of interest, and the extent of kin-selected altruism in the evolution and maintenance of cooperative breeding in this species.

Infanticide and cannibalism in wild common marmosets.

Infanticide has been observed in many different species [1], including common marmosets [2-4], due to sexual selection, reproductive strategies or resource competition [3, 5, 6], which may ultimately lead to exploitation (cannibalism) [1, 7]. Wild callithrichids have a very flexible mating system, including monogamy, polygynandry, polyandry and polygyny [4, 8, 9], with Monteiro da Cruz [10] finding all these patterns within the same population. This results from the high degree of deforestation of their habitat [4], but non-monogamous groups cannot ensure successful rearing of infants, since helpers are crucial and should be present in high numbers [11]. In this study, we show for the first time that cannibalism can follow infanticide, and we hypothesise that it is a result of both competition for scarce resources and the need for animal protein, exacerbated by forest degradation.

Excited levels in 149Pm from the decay of 149Nd.

The level structure of 149Pm has been investigated by studying the gamma rays emitted following the beta(-) decay of 149Nd (T(1/2)=1.7h). The singles and the gammagamma coincidence spectra were taken using HPGe detectors with high energy resolution. The energy and relative intensities of 198 gamma rays have been determined, 45 for the first time and several multiplets were resolved using bidimensional data analysis. A decay scheme with 51 levels has been proposed. This includes 6 new levels, at 1407, 1368, 1364, 1329, 1293 and 1181keV. The present results permitted assignments of spin and parity for a number of these levels.

Energy levels in 139La from the decay of 139Ba.

The gamma-ray spectrum of 139La following the beta- decay of 139Ba has been studied using both singles and gammagamma coincidence spectroscopy techniques. The energies and intensities of 30 gamma-transitions have been determined, which include three new transitions placed in the level scheme. Two new levels at 1524.6 and 1900.3 keV excitation energies are proposed and a number of gamma-transitions have been confirmed. On the basis of these results a precise decay scheme is proposed.

Decay of 72Ga.

The gamma-ray spectrum of 72Ge following the beta- decay of 72Ga has been studied using both single and gamma-gamma coincidence spectroscopy techniques. The energies and intensities of 110 gamma-rays have been determined, 26 of them were observed for the first time and 20 have been confirmed. Of the total number of gamma-rays observed, 95 have been placed in a proposed level scheme containing 31 levels. This includes five new levels at 2303, 2694, 3067, 3097 and 3420 keV.

Plasmodium falciparum: a comparative analysis of the genetic diversity in malaria-mesoendemic areas of Brazil and Madagascar.

For a better definition of the polymorphic features of Plasmodium falciparum parasite populations, the polymerase chain reaction (PCR) typing technique was used to investigate the genetic diversity and complexity of parasites harbored by acute P. falciparum carriers from three yet unexplored malaria-mesoendemic areas with different transmission levels: two localities in northwestern Brazil (Ariquemes and Porto Velho) and a village in Madagascar (Ankazobe). A total of 89 DNA samples were analyzed by amplification of polymorphic domains from genes encoding merozoite surface antigens 1 and 2 (MSP-1, MSP-2) and thrombospondin-related anonymous protein (TRAP) and by hybridization with allelic-family-specific probes or random-fragment-length polymorphism (RFLP). In all three localities, extensive polymorphism was observed for each marker, but the MSP-2 central repeat was the most diverse one. Similar levels of genetic diversity, allelic frequency, and infection complexity were observed in the two Brazilian localities, although the isolates had been sampled at 2-year intervals, suggesting the stability of the infecting parasite populations presenting in these regions of the Brazilian Amazon. Unexpectedly, although the entomologic inoculation rate was at least 3 times lower in Ankazobe than in the Brazilian areas. Malagasi samples appeared more complex than the Brazilian ones. The implications of these data with regard to parasite population-dynamics studies are discussed.

Plasmodium berghei: cerebral malaria in CBA mice is not clearly related to plasma TNF levels or intensity of histopathological changes.

Plasmodium berghei ANKA infection in CBA/J mice leads to the development of cerebral malaria (CM) that kills 80-90% of the animals in 6-9 days. This model has been used to study the pathogenesis of CM, which is a major cause of morbidity and mortality in Plasmodium falciparum-infected individuals. The role of cytokines in the induction of CM in the murine model has been well documented, but most studies have been restricted to the peak of neurological manifestations. Here we used a sequential approach to compare mice that developed CM with those that developed no cerebral pathology. Animals were examined for systemic histopathological changes and plasma Tumor Necrosis Factor-alpha (TNF) levels. The objectives were (a) to further determine the importance of factors commonly associated with murine CM-such as elevated levels of TNF and the presence of hemorrhage and vascular plugging-by comparing mice at different stages of infection and/or with different outcomes following infection and (b) to examine the importance of systemic changes-course of parasitemia and histopathological alterations in brain, liver, and lungs-in the development of CM. The data suggest that (a) the clinical manifestation of CM appears to be associated with a wave of merozoite release on days 6-7, (b) murine CM does not present reliable histopathological indicators, (c) there is no topographic association between the occurrence of intravascular plugging and the hemorrhagic foci, (d) monocyte-monocyte and monocyte-endothelial cell adherence were the most expressive histopathological events and were not restricted to brain vessels, (e) blood levels of TNF are not indicative of the local tissue reaction, (f) adhesiveness of monocyte/endothelial cells fluctuate during infection and is dissociated from the lymphocyte homing to the liver, and (g) pulmonary megakaryocytosis (megakaryopoiesis?) is a late event in the lungs.

The intraperitoneal Plasmodium berghei-Pasteur infection of Swiss mice is not a system that is able to detect the antiplasmodial activity in the Pothomorphe plant extracts that are used as antimalarials in Brazilian endemic areas.

The antimalarial activity of the hexane and methanol extracts derived from the Brazilian plants Pothomorphe peltata and Pothomorphe umbellata-whose leaves are popularly employed in medicinal folk remedies for the treatment of malaria-was assessed through in vivo tests with the Peters method. The extracts were delivered to Plasmodium berghei-infected mice via the oral or the subcutaneous route. A suppressive effect on the parasitemia seemed to be evident when data regarding the intraperitoneal injection of Pothomorphe umbellata extracts were analyzed. However, a definitive conclusion on an effective antimalarial activity is not possible, as two distinct-"standard" and "slow"-patterns of parasitemia occurring at similar frequencies in both treated and untreated intraperitoneally infected mice were observed. Nevertheless, the existence of two distinct profiles of parasitemia was not clear among the animals that were infected via the intravenous route. These data indicate the need for further studies on the biological features of the host/parasite interaction in the intraperitoneally P. berghei-infected Swiss mice system to standardize the model and to improve its usefulness in the screening of antimalarial compounds.

Cellular and antibody responses to the Plasmodium falciparum heat shock protein Pf72/HSP70 during and after acute malaria in individuals from an endemic area of Brazil.

Proliferative and antibody responses to three synthetic peptides corresponding to Pf72/ HSP70 were followed-up in acute malaria patients from an endemic area of Brazil. In vitro lymphocyte responsiveness to all peptides was relatively low and short-lived and there was a considerable variation in the frequency and magnitude of the individual lymphoproliferative response to the peptides at different periods after the onset of infection. Although 96% of the patients had IgG antibodies to crude Plasmodium falciparum asexual blood stage antigens, specific IgG antibody responses to the peptides varied from 12.5 to 40% according to the tested peptides. No significant difference was observed in the proliferative or antibody responses to the peptides between individuals that remained parasitemic after treatment and those that recovered from malaria infection. The different frequencies of proliferative responses in peripheral blood T cells on different occasions after the onset of their infection show that, in order to be informative, evaluation of the in vitro cellular immune response to peptides requires longitudinal studies in which each individual is tested repeatedly.

A study of antibody and T cell recognition of rhoptry-associated protein-1 (RAP-1) and RAP-2 recombinant proteins and peptides of Plasmodium falciparum in migrants and residents of the state of Rondonia, Brazil.

Humoral and cellular responses were examined among natives and migrants in an area of the Amazon region of Brazil. Rhoptry-associated protein-1 (RAP-1) and RAP-2 expressed in Escherichia coli expression systems, a peptide corresponding to the epitope bound by inhibitory anti-RAP-1 antibodies, and four other RAP-1 and RAP-2 synthetic peptides were used in these studies. Plasma from the native population had greater IgG reactivity to the N-terminal third of RAP-1 than the migrant population; both populations had low levels of IgM to this region of RAP-1. The IgG reactivity to RAP-2 and to the C-terminal third of RAP-1, as well as for all the peptides, including the peptide from the inhibitory domain, were low or absent in both populations. In contrast, there were a high number of subjects with an IgM response to the peptides. Cellular responses were measured by proliferation of peripheral blood mononuclear cells (PBMC) and, in some subjects, by reverse transcription-polymerase chain reaction for interleukin-2 (IL-2), interferon-gamma (IFN-gamma), IL-4, and IL-10. Proliferation of PBMC was low when stimulated by recombinant proteins, peptides, or parasite lysate. Both RAP-1 and RAP-2 stimulated cytokine production by donor T cells; IL-2, IL-4, and IFN-gamma RNA transcripts were observed in response to recombinant proteins and parasite lysate, but with no uniform trends. From the observed antibody responses, RAP-1 appears to be more immunogenic than RAP-2.

Seroepidemiology of abdominal angiostrongyliasis: the standardization of an immunoenzymatic assay and prevalence of antibodies in two localities in southern Brazil.

Abdominal angiostrongyliasis is a nematode disease produced by Angiostrongylus costaricensis, a metastrongylid parasite of wild rodents. Accidental human infection occurs through ingestion of food or water contaminated with third-stage larvae present in the mucous secretion of terrestrial molluscs. An ELISA test was standardized for detection of IgG antibodies recognizing a surface antigen prepared from female worms. Competitive absorption of sera with Ascaris suum crude antigen resulted in a test with 86% sensitivity and 83% specificity. The disease is endemic in Southern Brazil and a number of cases are diagnosed every year through anatomo-pathological examination of biopsies or surgical specimens, since no other diagnostic method is available. According to seroepidemiological studies, prevalences in two transmission foci are 29.8 and 66%, attesting to the widespread occurrence of the infection in those endemic areas.

Can software be used to predict antigenic regions in Plasmodium falciparum peptides?

We compared the antigenicity of p126 Plasmodium falciparum peptides with predicted antigenic regions identified using the methods described by Garnier et al. (1978) and Chou & Fasman (1974). For this purpose nine different P. falciparum peptides were synthesized in accordance with the deduced amino acid sequence of the p126 gene, and their reactivity was tested using an enzyme linked immunosorbent assay against sera from individuals with a natural malaria infection. Both predictive methods gave similar antigenic-index scores, however, a comparison of these predictive results with data obtained by ELISA showed that the probability of a correct prediction was only around 45% for both cases. Thus, in our view computer software could not be used in isolation for screening purposes, and other parameters must also be taken into account when using such software to assess antigenicity.