Fasciola hepatica in Brazil: genetic diversity provides insights into its origin and geographic dispersion.

Fasciola hepatica is a trematode parasite that affects mammals, including humans. In Brazil, fascioliasis, a disease caused by the parasite, is of great importance. The disorder affects the welfare of the Brazilian population through impairing the agricultural production of cattle, where the disease causes weight loss as a result of liver damage. This study aimed to evaluate the genetic diversity of F. hepatica throughout Southern Brazil to determine its geographic origin and estimate the colonization route of the parasite. To accomplish these aims, flukes were collected from slaughterhouses in three endemic areas of Rio Grande do Sul and Paraná states. DNA was isolated using the phenol-chloroform protocol from single flukes and two mitochondrial genes, cytochrome oxidase subunit I (COI) and nicotinamide dehydrogenase subunit 1 (Nad1), were amplified and sequenced. Ten haplotypes of COI were found from 75 isolated parasites and the total haplotype and nucleotide diversity observed were 0.475 and 0.002, respectively. Using the Nad1 gene, we found 24 haplotypes from 79 samples, resulting in haplotype and nucleotide diversity values of 0.756 and 0.004, respectively. An analysis of molecular variance showed that 57.4% and 77.5% of variation was within populations (FST), while 9.0 and 36.8% of variation was among groups (FCT) when considering COI and Nad1 genes, respectively. For COI, the fixation index values of 0.425 and 0.368 were obtained for FST and FCT, respectively, while analysis of Nad1 0.225 and 0.089 index values were obtained for FST and FCT, respectively. We have determined that F. hepatica found in the two distinct areas originated from several geographical regions, since we found haplotypes that were shared with at least three different continents. These data are in accordance with the recent colonization of Brazil, and the recent import of cattle from South American, European and, possibly, some African countries. The observed FST and FCT values for COI and Nad1 genes of F. hepatica may be a result of limited movement of animals within states and support the lack of geographical structure of the parasite in Brazil, which are in agreement with the observed cattle production systems in this region.

Echinococcus canadensis (G7) and Echinococcus granulosus sensu stricto (G1) in swine of southern Brazil.

The cystic echinococcosis (CE) is an important zoonotic disease caused by the parasite Echinococcus spp. In Brazil, this parasite is present in Rio Grande do Sul (RS) state, border with Argentina and Uruguay, causing several damages to human and animal health. This study aimed to identify Echinococcus spp. in hydatid cysts of swine and evaluate the similarity of the genotypes through the phylogenetic analysis. A total of 3,101,992 swine were slaughtered in the central/northern region of RS/Brazil, during 2008-2012. Five isolates were characterized as hydatid cyst by molecular analysis, based on the mitochondrial gene cytochrome c oxidase subunit I (cox-I). The genotypes E. granulosus sensu stricto (G1) (n=2) and E. canadensis (G7) (n=3) were identified in the hydatid cysts. The swine represents a potential intermediate host for different genotypes of Echinococcus spp., besides it can contribute to the perpetuation of the parasite's life cycle in rural areas.

Reappraising the theme of breeding systems in Echinococcus: is outcrossing a rare phenomenon?

Selfing has been considered the most common mode of reproduction in Echinococcus flatworms. However, population genetic studies on the asexual larval stage involving nuclear co-dominant markers have not always revealed significant heterozygote deficiencies--the expected outcome of a regularly and highly inbred population. In this study, we analysed the genetic structure of Echinococcus granulosus sensu lato populations from Southern Brazil during their adult (sexual) stage using 1 mitochondrial and 1 nuclear marker (cox 1 and mdh, respectively). We show that parasite genetic differentiation is largest among definitive hosts (domestic dogs) from different farms, suggesting that transmission is mostly maintained within a farm. Moreover, we show that heterozygote deficiencies are not significant, and we suggest that outbreeding is the most common mode of reproduction of the parasite in that region.

Diphenyl diselenide [(PhSe)2] inhibits Drosophila melanogaster delta-aminolevulinate dehydratase (delta-ALA-D) gene transcription and enzyme activity.

The main objective of the present study was to compare the inhibitory effect of diphenyl diselenide (PhSe)(2) and Pb(2+) on mice and fruit fly delta-Aminolevulinate dehydratase (delta-ALA-D). Optimum pH was quite different for mice (pH 6.5) and flies (pH 8.5). At pH 8.5, the inhibitory potency of (PhSe)(2) was higher for the fruit flies (IC(50) 8.2 micromol/l) than for mice (IC(50) 19.5 micromol/l). Pb(2+) inhibited mice delta-ALA-D at pH 6.5 (IC(50) 6.2 micromol/l) and 8.5 (IC(50) 5.6 micromol/l) with higher potency than the fly enzyme (IC(50) 43.7 micromol/l). delta-ALA-D transcription was reduced by 15% in flies exposed to 0.3 mmol/kg (PhSe)(2), which is similar to the reduction observed in activity measured in the presence of dithiothreitol. The three-dimensional prediction by SWISS-PROT mouse and fly delta-ALA-D revealed differences in the number of hydrogen bonds and turns for the 2 enzymes. Sulfhydryl groups (-SH) that could be oxidized by (PhSe)(2) are conserved in the two sources of enzyme. Distinct responsiveness to pH, (PhSe)(2) and Pb(2+) of these enzymes may be related to subtle differences in tertiary or quaternary structure of mouse and fly delta-ALA-D. Furthermore, mechanism underlying enzyme inhibition after in vivo exposure seems to be different for Drosophila melanogaster and rodent enzymes.

Expression and diversity of Echinococcus multilocularis AgB genes in secondarily infected mice: evaluating the influence of T-cell immune selection on antigenic variation.

The T-cell-mediated immune response exhibits a crucial function in the control of the intrahepatic proliferation of Echinococcus multilocularis larvae in mice and humans, both being natural intermediate hosts of the parasite. Antigen B (AgB), a metabolized Echinococcus spp. lipoprotein, contributes to the modulation of the T-cell immune response, and distinct sites of the corresponding AgB1, AgB3 and AgB4 genes were shown to be under positive selection pressure. Since several AgB gene variants are present in a single Echinococcus metacestode, we used secondary E. multilocularis infections in BALB/c and in athymic nude mice (devoid of T-cell responses) to analyze the effect of the cellular immune response on the expression and diversity of EmAgB1-EmAgB4 genes. We demonstrated hereby that EmAgB transcripts were less abundant in nude mice during the early phase of infection (at one month post-infection), and that EmAgB2 is simultaneously down-regulated when compared to the other three genes. A negative relationship exists between the level of transcription and diversity of EmAgB genes. Moreover, no excess of non-synonymous substitutions was found among the distinct EmAgB alleles from a single host. Together, these results pointed to the effect of purifying selection, which seemed to eliminate the detrimental AgB variants generated during the development of the metacestode within the peritoneal cavity of its intermediate host.

Clinicopathologic correlation of spontaneous retinal pigment epithelial tears with choroidal neovascular membranes in age-related macular degeneration.

BACKGROUND: A correlation of the clinical and histopathologic features of age-related macular degeneration (AMD), complicated by choroidal neovascular membrane (CNVM) with spontaneous tears of retinal pigment epithelial (RPE) detachment, has not been reported. METHODS: Two patients with vision loss from AMD and spontaneous pigment epithelial detachment tears, involving the fovea with serous or hemorrhagic neurosensory retinal detachments, were examined clinically. The surgically excised CNVM from each patient was examined with light microscopy. RESULTS: The authors present two patients with clinicopathologic correlation of the ophthalmoscopic, fluorescein angiographic, and pathologic findings of spontaneous pigment epithelial detachment tears. In the surgical specimens from these two eyes, the removed tissue, presumed to be the spontaneous RPE tear with CNVMs, demonstrated a fibrovascular membrane surrounding RPE with adjacent abnormally thickened basement membrane. The RPE-basement membrane complex was folded redundantly and scrolled beneath itself. CONCLUSION: The authors found RPE-basement membrane abnormalities and CNVMs associated with clinically evident spontaneous pigment epithelial detachment tears. These findings support the current theories of their pathogenesis.

kappa-Immunoglobulin corneal deposits associated with monoclonal gammopathy. Immunohistochemical and electron microscopic findings.

A 62-year-old man had a 1-year history of bilaterally decreased visual acuity associated with irregular, patchy, grayish-white stromal infiltrates involving both corneas. As visualized by light microscopy, the keratectomy specimen showed numerous homogeneous, eosinophilic deposits located mostly within keratocytes in the posterior one-third of the stroma. The deposits stained reddish-brown with Masson's trichrome but were periodic acid/Schiff-negative. As visualized by electron microscopy, the intracytoplasmic deposits were located within dilated cisternae of rough-surfaced endoplasmic reticulum of keratocytes and disclosed a 10-nm periodicity with a distinct lattice pattern. Immunoperoxidase strains confirmed that the deposits contained immunoglobulin (IgG-kappa). At 2 weeks following a histopathologic examination of the keratectomy specimen, serum protein analysis and immunoelectrophoresis with quantitation of immunoglobulins demonstrated an IgG-kappa monoclonal gammopathy. The abnormal corneal deposits may be the first clinical sign of a malignant lymphoproliferative process that may be associated with a monoclonal gammopathy.

[On the question of a diurnal rhythm of the arterial oxygen tension (pO2a) (author's transl)]. / Zur Frage einer Tagesrhythmik des arteriellen Ruhe-pO2a.

In connection to our clinical examination of the so-called "Oxygen-Multistep-Therapy" (02-MT) was to clear, if the pO2a could be affected with a diurnal rhythm over the daytime between 7 a.m. and 5 p.m. The results, which were found at 66 healthy persons of different age and sex show an independence of the pO2 measured values from the daytime. Further, the action of Finalgon upon the earlap, and the variation of pO2a in dependence on age and sex were established, and a correlation between the arterial blood pressure and pO2a is discussed.

[Glucose and K+ balance during high-dosage intravenous glucose infusion]. / Glukose- und K+-Bilanz während hochdosierter intravenöser Glukosezufuhr.

Intravenous infusion of glucose in high dosage was given to 42 tumor patients, attaining a hyperglycemia of about 40 mg/100 ml over a period of 24 hours. This hyperglycemia is a part of the so-called "cancer-multistep-therapie". During the period of glucose-infusion a water-turnover of about 6,000 ml can be registered which is connected with a requirement of glucose of about 2,000 g. Only 1/3 of this amount is loosing by renal excretion, but 2/3 are metabolised to correspond with a glucose-uptake of 0,94 g and a glucosuria of 0,32 g per kg bodyweight and hour. Requirement of glucose and glucose uptake are in inverse proportion to the age of the patients and to the duration of hyperglycemia. Further a "glucose-potassium-equivalent" existed: about 10 mval potassium per 100 g infused glucose, whereas the netto uptake of potassium is 0,049 mval per g of metabolised glucose. Acid-base-equilibrium and red blood-picture did not show any relevant variations while typical deviations of some other parameters (cardiac-frequency, body-temperature), white blood-picture) are can be associated with the so-called "general adaptation syndrom". The findings are discussed in view of their general importance for the parenteral application of glucose.

[Clinical examination of the conception '86 of "Canoer Multistep-Therapie" (CMT) by M. v. Ardene. First communication: the standard, the control-parameters, and the course of therapy (author's transl)]. / Klinische Prüfung des Krebs-Mehrschritt-Therapie-(KMT)-Konzepts '74 nach M. v. Ardenne. I. Mitteilung. Therapiestandard, Therapieüberwachungsparameter und Therapieverlauf

It is reported about the clinical testing of the "Cancer Multistep Therapy" (CMT) by M. von Ardenne (CMT Concept '74). It was made in 1974/75 on 42 tumour-patients. At first we demonstrate in this publication clinical and paraclinical findings, which may be relevant for the therapy-standard, but the results of the therapy-efficiency will be given later. On the base of findings and observations we verify a program-true employment of the CMT Concept '74 and we demonstrate the pathophysiological reaction-patterns in connection with the longterm-hyperglycemia (glucose-utilization, spontaneous hyperthermy with tachycardia, spontaneous leukocytosis with typical deviations in the hematogram). These reaction-patterns are described particularly as "general adaptation syndrome" (Selye). Further we give an assessment of compatibility, tolerableness, and practicability of the CMT.