Observations on the distribution of anophelines in Suriname with particular reference to the malaria vector Anopheles darlingi

A study was made on the distribution of anophelines in Suriname with special emphasis on the principal malaria vector Anopheles darlingi and on the occurrence of other possible vector species. Peridomestic human bait collections of adult mosquitoes and collections of larvae were made in many localities with a recent history of malaria transmission. Stable population of An. darlingi were only found in the interior, south of the limit of tidal influence, due to year-round availability of breeding habitats in quietly sunlit places in flooded forest areas and along river banks. In the area with tidal movement of the rivers, breeding is limited to flooded areas in the west season. Anopheles darlingi was only incidentally collected in low densities. In the interior, malaria transmission occurred in all places where An. darlingi was found. The absence of malaria transmission along the Upper Suriname River could be explained by the absence of An. darlingi. In the malaria endemic areas, An darlingi was the most numerous mosquito biting on man. In the tidal region, malaria outbreak are infrequent and might be explained by the temporary availability of favourable beeding habitats for An. darlingi. However, evidence is insufficient to incriminate an. darlingi as the vector of malaria in this region and the possible vectorial role of other anophelines is discussed.

Trypanosoma rangeli (Tejera, 1920): observations upon pleomorphism

Metatrypomastigotes of Trypanosoma rangeli Tejera, 1920, harvested from LIT medium, were inoculated i.p. or s.c. into 6, 16, and 26g NMRI mice, these representing increasing degrees of immunological maturity. In all cases, similar pleomorphic patterns were observed. Four morphobiometrically differentiable types of trypanosome were encountered in an overlapping temporal sequence. These observations, taken in comparison with those on pleomorphism in this and other species of Trypanosoma by other workers, are consistent with the hypothesis that the pleomorphic types represent the natural development of the parasite, rather than the result of the immune response of the mammal host. Small, slender trypanosomes prevalent at the onset of the parasitemia either reinvade the tissue cells for relatively limited subsequent generations of tissue reproduction, or else differentiate toward the forms that are only capable of colonizing the insect vector.

Replication of dengue viruses in mosquito cell cultures: a model from ultrastructural observations

Mosquito cell cultures infected with human sera from dengue-1 and dengue-2 outbreaks, started in Rio de Janeiro by 1986 and 1990 respectively, were examined by electron microscopy at different times post the infection of cell cultures. More information was obtained about cell penetration of virus particles in the presence or not of antibodies, their pathway inside the cells, replication mode and exit. Infectiveness of the virus at those different stages can only be attributed to the particles appearing inside the trans-Golgi vesicles; most of all newly formed virus particles remain inside the RER-derived cell vesicles or inside lysosomes, even during cell lysis. Groups of larges particles, 65-75 nm in diameter at dengue-2 infections, persist during cell passage. The large amounts of smooth membrane structures, as vesicles or tabules inside the RER are attributed to a cell response to viral infection.

Context-dependent interpretation of the prognostic value of BRAF and KRAS mutations in colorectal cancer.

BACKGROUND: The mutation status of the BRAF and KRAS genes has been proposed as prognostic biomarker in colorectal cancer. Of them, only the BRAF V600E mutation has been validated independently as prognostic for overall survival and survival after relapse, while the prognostic value of KRAS mutation is still unclear. We investigated the prognostic value of BRAF and KRAS mutations in various contexts defined by stratifications of the patient population. METHODS: We retrospectively analyzed a cohort of patients with stage II and III colorectal cancer from the PETACC-3 clinical trial (N = 1,423), by assessing the prognostic value of the BRAF and KRAS mutations in subpopulations defined by all possible combinations of the following clinico-pathological variables: T stage, N stage, tumor site, tumor grade and microsatellite instability status. In each such subpopulation, the prognostic value was assessed by log rank test for three endpoints: overall survival, relapse-free survival, and survival after relapse. The significance level was set to 0.01 for Bonferroni-adjusted p-values, and a second threshold for a trend towards statistical significance was set at 0.05 for unadjusted p-values. The significance of the interactions was tested by Wald test, with significance level of 0.05. RESULTS: In stage II-III colorectal cancer, BRAF mutation was confirmed a marker of poor survival only in subpopulations involving microsatellite stable and left-sided tumors, with higher effects than in the whole population. There was no evidence for prognostic value in microsatellite instable or right-sided tumor groups. We found that BRAF was also prognostic for relapse-free survival in some subpopulations. We found no evidence that KRAS mutations had prognostic value, although a trend was observed in some stratifications. We also show evidence of heterogeneity in survival of patients with BRAF V600E mutation. CONCLUSIONS: The BRAF mutation represents an additional risk factor only in some subpopulations of colorectal cancers, in others having limited prognostic value. However, in the subpopulations where it is prognostic, it represents a marker of much higher risk than previously considered. KRAS mutation status does not seem to represent a strong prognostic variable.

Pédiatrie. 4. La calprotectine fécale en pédiatrie: utilisation et interprétation [Pediatrics. Fecal calprotectin in children: use and interpretation].

Fecal calprotectin is a small protein released mainly by neutrophils. It is recognized as a reliable, easy and non-invasive biomarker of gastro-intestinal inflammation. Normal values vary with age, with higher cut-off values during the first year of life (<350 microg/g) than in children (<275 microg/g) or adults (<50 microg/g). Fecal calprotectin can be a useful tool in initial evaluation of recurrent abdominal pain, helping to distinguish between functional gastro-intestinal disorders, where it is normal, and inflammatory bowel disease (IBD). It is not a specific marker of IBD but is increased in other situations of gastro-intestinal inflammation. In patients with IBD, fecal calprotectin is used to monitor treatment response.

Visceral leishmaniasis in Teresina, state of Piauí, Brazil: preliminary observations on the detection and transmissibility of canine and sandfly infections

A Leishmania donovani-complex specific DNA probe was usedto confirm the widespread dissemination of amastigotes in apparently normal skinof dogs with canine visceral leishmaniasis. When Lutzomyia longipalpis were fed on abnormal skin of five naturally infected dogs 57 of 163 (35 per cent) fliesbecame infected: four of 65 flies (6 per cent) became infected when fed on apparently normal skin. The bite of a single sandfly that had fed seven days previouslyon a naturally infected dog transmitted the infection to a young dog from a non-endemic area. Within 22 days a lesion had developed at the site of the infectivebite (inner ear): 98 days after infection organisms had not disseminated throughout the skin, bone marrow, spleen or liver and the animal was still serologically negative by indirect immunofluorescence and dot-enzyme-linked immunosorbent assay. When fed Lu. longipalpis were captured from a kennel with a sick dog known to be infected, 33 out of 49 (67 per cent) of flies contained promastigotes. In contrast only two infections were detected among more than 200 sandflies captured in houses. These observations confirm the ease of transmissibility of L.chagasi from dog to sandfly to dog in Teresina. It is likely that canine VL is the major source of human VL by the transmission route dog-sandfly-human. the Lmet2 DNA probe was a useful epidemiological tool for detecting L. chagasi in sandflies.

Human papillomavirus detection in genital lesions by in situ hybridization and ultrastructural observations

Detection of papillomavirus DNA in sity hybridization technique was perfomed in 29 symptomatic patients (6 males and 23 females) during the period of 1989-1991 at the Clinic for Sexually Transmitted Diseases, Universidade Federal Fluminense, State of rio de Janeiro. All the male patients had condyloma acuminata. Only HPV 6/11 were found in these lesions. Clinical features inthe female patients included vulvar condyloma acuminata, bowenoid populosis, flat cervical condyloma, cervical condyloma acuminatum and cervical intraepithelialneoplasia grade II (CIN II). We also found cases of condyloma acuminata associated to vulvar intraepithelial neoplasia grade III (VIN III), as well as to vaginal invasive carcinoma. HPV 6/11 and 16/18 were found in vulvar condyloma acuminata. Mixed infection by 6/11-16/18 HPV were also seen in these lesions as well as in the patient who had cervical condyloma acuminatum. HPV 16/18 were found in the condyloma acuminatum plus VIN III and in the CIN II lesions. We have found HPV31/33/51 in the specimen of condyloma acuminatum plus invasive carcinoma. In order to investigate the ultrastructural aspects of HPV infection in genital tissue, the biopsies of three female patients were observed under electron microscope.Mature virus particles were found in the cells of a condyloma acuminatum as wellas in the condyloma acuminatum plus invasive carcinoma case. In another sample, chromosome breakages were found in the nuclei of the infected cells although no viral particles were observed.

Observations on some Avian Coccidia (Apicomplexa: Eimeriidae) in Amazonian Brazil

Oocysts of Eimeria porphyrulae n. sp. are described in faeces of Porphyrula martinica (Aves: Gruiformes: Rallidae). They are ellipsoidal to oval, 22.4 x 17.7 (20.0-23.7 x 16.2-18.7) µm, shape-index (length/width) 1.3. Oocyst wall about 1.25 µm thick, colourless, with two layers: inner one prominently striated. Micropyle and sub-micropylar granule present: no oocyst residuum. Sporocysts 17.5 x 9.0 (17.0-19.0 x 8.0-10.0) µm, shape-index 1.9, with inconspicuous Stieda/sub-Stieda bodies. Sporocyst residuum of scattered granules, sometimes a compact mass: sporozoites with two refractile bodies. Eimeria crypturelli n. sp. is described in faeces of Crypturellus soiu (Tinamiformes: Tinamidae). Oocysts ellipsoidal-oval, 20.75 x 14.5 (17.5-25.0 x 11.25-21.25) µm, shape-index 1.4. Oocysts wall about 1.25 µm thick and bi-layered: inner layer faintly striated. Micropyle present, with oocyst residuum immediately below: single polar body rarely present. Sporocysts 13.0 x 7.5 (12.5-13.75 x 7,5-8.1) µm, shape-index 1.7, with a Stieda body but seemingly no sub-Stieda. Sporocyst residuum compact: sporozoites with two refractile bodies. Isospora cacici n. sp. is recorded from faeces of Cacicus cela cela (Passeriformes: Icteridae). Oocysts subspherical-spherical, 26.5 x 23.7 (22.5-27.5 x 20.0-26.2) µm, shape-index 1.1. Wall a single, colourless layer about 1.5 µm thick. No micropyle or oocyst residuum: 1-2 polar bodies. Sporocysts ellipsoidal, 17.7 x 12.5 (17.5-18.75 x 11.25-13.75) µm, shape-index 1.4, with pronounced Stieda/sub-Stieda bodies: residuum compact and sporozoites with two refractile bodies. Isospora thraupis n. sp. is described from faeces of Thraupis palmarum melanoptera (Passeriformes: Thraupidae). Oocysts subspherial-spherical, 19.9 x 19.0 (18.7-21.2 x 18.75-20.0) µm, shape-index 1.0. Wall about 0.6 µm thick, smooth, colourless and a single layer: no micropyle, oocyst residuum or polar bodies. Sporocysts 14.2 x 9.2 (13.7-16.2 x 8.7-10.0) µm, shape-index 1.5: Stied/sub-Stieda bodies inconspicuous. Residuum compact: sporozoites with two refractile bodies.

Ultrastructural Observations of the Vitelline Cells of Metamicrocotyla macracantha (Monogenea, Microcotylidae)

An electron microscopic study of the vitelline follicles of Metamicrocotyla macracantha (Alexander, 1954) Koratha,1955 showed that they are composed of cells in different stages of development. The immature cells have a large nucleus, nucleolus, cytoplasm with free ribosomes and few mitochondria. The developing vitelline cells present granules which are small in the early stages, increasing with maturity. The mature cells have an extensive granular endoplasmic reticulum and droplets of shell-protein; with maturation, clusters of shell protein and yolk bodies are formed and released in the ciliated vitelline ducts. Vitelline development is continuous and all of the cellular stages involved can be found in each follicle.

RTOG 0525: Exploratory Subset Analysis from a Randomized Phase III Trial Comparing Standard (std) Adjuvant Temozolomide (TMZ) with a Dose-dense (dd) Schedule for Glioblastoma (GBM)

Purpose/Objective(s): RTwith TMZ is the standard for GBM. dd TMZ causes prolongedMGMTdepletion in mononuclear cells and possibly in tumor. The RTOG 0525 trial (ASCO 2011) did not show an advantage from dd TMZ for survival or progression free survival. We conducted exploratory, hypothesis-generating subset analyses to detect possible benefit from dd TMZ.Materials/Methods: Patients were randomized to std (150-200 mg/m2 x 5 d) or dd TMZ (75-100 mg/m2 x 21 d) q 4 weeks for 6- 12 cycles. Eligibility included age.18, KPS$ 60, and. 1 cm2 tissue for prospective MGMTanalysis for stratification. Furtheranalyses were performed for all randomized patients (''intent-to-treat'', ITT), and for all patients starting protocol therapy (SPT). Subset analyses were performed by RPA class (III, IV, V), KPS (90-100, = 50,\50), resection (partial, total), gender (female, male), and neurologic dysfunction (nf = none, minor, moderate).Results: No significant difference was seen for median OS (16.6 vs. 14.9 months), or PFS (5.5 vs. 6.7 months, p = 0.06). MGMT methylation was linked to improved OS (21.2 vs. 14 months, p\0.0001), and PFS (8.7 vs. 5.7 months, p\0.0001). For the ITT (n = 833), there was no OS benefit from dd TMZ in any subset. Two subsets showed a PFS benefit for dd TMZ: RPA class III (6.2 vs. 12.6 months, HR 0.69, p = 0.03) and nf = minor (HR 0.77, p = 0.01). For RPA III, dd dramatically delayed progression, but post-progression dd patients died more quickly than std. A similar pattern for nf = minor was observed. For the SPT group (n = 714) there was neither PFS nor OS benefit for dd TMZ, overall. For RPA class III and nf = minor, there was a PFS benefit for dd TMZ (HR 0.73, p = 0.08; HR 0.77, p = 0.02). For nf = moderate subset, both ITT and SPT, the std arm showed superior OS (14.4 vs. 10.9 months) compared to dd, without improved PFS (HR 1.46, p = 0.03; and HR 1.74, p = 0.01. In terms of methylation status within this subset, there were more methylated patients in the std arm of the ITT subset (n = 159; 32 vs. 24%). For the SPT subset (n = 124), methylation status was similar between arms.Conclusions: This study did not demonstrate improved OS for dd TMZ for any subgroup, but for 2 highly functional subgroups, PFS was significantly increased. These data generate the testable hypothesis that intensive treatment may selectively improve disease control in those most likely able to tolerate dd therapy. Interpretation of this should be considered carefully due to small sample size, the process of multiple observations, and other confounders.Acknowledgment: This project was supported by RTOG grant U10 CA21661, and CCOP grant U10 CA37422 from the National Cancer Institute (NCI).