[Dengue virus infection in neotropical forest mammals: incidental hosts or potential reservoirs?]. / Infection par le virus de la dengue de mammifères sauvages en région néotropicale: hôtes accidentels ou réservoirs potentiels?

The arboviral disease with the highest human incidence in South America is dengue fever. In French Guiana, where all four dengue serotypes, i.e., DENV-1, DENV-2, DENV-3 and DENV-4, are present, the disease is endemic with epidemic outbreaks. Though previous serological studies have suggested a sylvatic cycle, involvement of wild mammals in the dengue cycle in the neotropics has never been confirmed. The purpose of this study was to search for the presence of DENV in wild animals captured at two different sites between 2001 and 2007. About 10,000 trap/nights were performed leading to the capture of 464 non-flying mammals (rodents and marsupials). In addition, mistnests placed in the same zone yielded 152 bats. Reverse transcription-polymerase chain reaction amplification to detect infection by any of the four dengue serotypes demonstrated viral RNA in the livers and/or sera of 92 captured animals. Sequence analysis of amplification products revealed that the DENV-1, DENV-3 and DENV-4 serotypes were distinct from those circulating in humans at the same periods. Analysis for DENV-2 showed that some strains were divergent from concurrent human strains but that others were identical. The latter finding suggests that wild neotropical mammals living in periurban area can be infected by dengue virus strains circulating in humans. However, further investigation will be needed to determine if neotropical mammals are incidental hosts or potential reservoirs of dengue virus.

[Genetic diversity and evolution of primate Gammaherpesvirinae]. / Diversité génétique et évolution des Gammaherpesvirinae de primates.

The Gammaherpesvirinae sub-family is divided into two genera, the Lymphocryptovirus and the Rhadinovirus. Until recently, Epstein-Barr virus (EBV), the human prototype of the Lymphocryptovirus genus, and simian homologues have only been detected in humans and Old World non-human primates. In other respects, the Rhadinovirus genus was only represented by Herpesvirus saimiri and Herpesvirus ateles of New World monkey species. Therefore, the general thinking at that time was that the separation of the continents resulted in drastic changes in the Gammaherpesvirinae evolution. The discovery of the human herpesvirus 8 (HHV8), belonging to the Rhadinovirus genus, followed by the identification of CalHV3 (Callitrichine herpesvirus 3) a lymphocryptovirus of marmoset, challenged this old paradigm. The recent description of numerous viruses belonging to the Gammaherpesvirinae subfamily from different Old and NewWorld primate species let to develop and to support co-speciational evolution hypotheses of these viruses and their hosts. This review focuses on our current knowledge of the genetic diversity and evolution of primate Gammaherpesvirinae.

Aortic 125I-albumin transport in patients with Marfan's syndrome and annuloaortic ectasia.

The morphologic, biochemical, and mechanical abnormalities of connective tissue fibrous proteins in Marfan's syndrome have been well studied, and their role in cardiovascular complications is well accepted. Less is known, however, about the state of the amorphous components of the aortic connective tissue. In the course of a study of transmural transport in blood vessels, we have had the opportunity to study dystrophic aorta from two young men who survived elective surgery; both with aortic insufficiency (AI) histologically compatible with Marfan's syndrome. One had recurrent chronic dissecting aneurysm (RCDA) as well. The aorta of the first (but not the second) was histologically compatible with Marfan's syndrome. Fresh specimens of intact ascending aorta were incubated in Krebs solution, pH 7.4, containing 125I-labelled bovine serum albumin for 2 h at 37 degrees C. The samples were then frozen, and serially sectioned in the plane of the lumenal surface. The radioactivity of the 20-micron thick sections was then determined, and expressed as a tissue/labelled solution concentration ratio. Transmural profiles of these ratios revealed no difference between the aorta of the RCDA patient with non-specific aortic dystrophy, and that of a 70-year-old man undergoing aortocoronary bypass. However, in the patient with aortic histology compatible with Marfan's syndrome, the average media concentration ratio was 5-fold less (4% vs. 20%).

A primary immunoblastic T malignant lymphoma of the small bowel, with azurophilic intracytoplasmic granules. A histologic, immunologic, and electron microscopy study.

We report an aggressive primary T-immunoblastic lymphoma of the small intestine without blood involvement or associated celiac disease. Grossly, the tumor was composed of multiple disseminated ulcerated, infiltrating, or protuberant nodular lesions. Immunologic investigation showed that lymphoma cells were of peripheral (post-thymic) T-cell origin and expressed the phenotype associated with cytotoxic-suppressor subset (Leu4/CD3+, Leu9/CD7+, Leu2/CD8+, Leu11/CD16+, Leu 7/NKcells+, FcIgG+, HLA-DR+, anti-Tac/CD25+, Ki-1/CD30-, Leu1/CD5-, Leu5/CD2-, Leu3/CD4-). A particular morphologic feature of this case is the presence of numerous azurophilic granules within the lymphoma cells, identified as lysosomes by cytochemical and ultrastructural studies. In view of recent immunologic evidence that normal cytotoxic/suppressor T-cells selectively reside within the epithelium of the normal bowel and some of them contain azurophilic granules, it could be suggested that our patient's lymphoma represents a malignant counterpart of these lymphocytes. Furthermore, the aggressive character of this T malignant lymphoma (T-ML) could be related to the expression of T-cell activation markers HLA-DR and Tac/CD25 and the proliferation-associated antigen Ki-67 on a high proportion of tumor cells.

Limitations of fluorocarbons in reducing myocardial infarct size.

The effects of the oxygen-carrier fluorocarbons on myocardial infarct size were assessed in non-exchange-transfused dogs subjected either to a 3-hour occlusion of the left anterior descending coronary artery (LAD) followed by 2 hours of reperfusion (protocol I) or to a 5-hour permanent LAD occlusion (protocol II). Fluorocarbon administration was begun 30 minutes after LAD occlusion and was continued over the entire period of ischemia. After 5 hours, the hearts were excised and areas of necrosis were visualized by triphenyl tetrazolium chloride staining while risk regions were assessed by radiolabeled microspheres injected after coronary occlusion just before the onset of therapy, and further, in protocol I, by thallium-201 perfusion imaging performed at the end of fluorocarbon administration. In protocol I experiments, the ratio of necrotic area to area at risk was 81 +/- 35% (mean +/- standard deviation) in control saline-treated dogs (n = 6) and 67 +/- 27% in fluorocarbon-treated dogs (n = 6) (difference not significant). There was no significant difference between risk regions measured after and before fluorocarbon treatment. In protocol II, the ratio of necrotic area to area at risk was 47 +/- 30% in control dogs (n = 5) and 63 +/- 29% in fluorocarbon-treated dogs (n = 5) (difference not significant). However, in control dogs, the ratio of necrotic area to area at risk increased from 47 +/- 30% in the dogs that underwent permanent occlusion to 81 +/- 35% in the group that underwent reperfusion (p less than 0.001) while this ratio was similar in the corresponding subsets of fluorocarbon-treated animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Reduction of myocardial infarct size by a fluorocarbon-oxygenated reperfusate.

This investigation assesses whether the size of an acutely revascularized myocardial infarct (MI) could be reduced by altering the composition of the initial reperfusate. Nineteen open-chest dogs underwent 4-hour occlusion of the left anterior descending coronary artery and were then assigned to a treatment group: 12 dogs to selective intracoronary infusion of the modified reperfusate over 30 minutes before resumption of blood flow for 60 minutes and 7 to a control group (90 minutes of unmodified blood reperfusion). The modified reperfusate consisted of 500 ml of a fluorocarbon-oxygenated crystalloid solution (PO2 650 mm Hg; total O2 content 5.5 vol%) whose composition was adjusted by decreasing Ca++ (0.25 mM), increasing pH (7.60) and adding glucose (1.8 g/liter). Four hours after occlusion, technetium-99m-labeled microspheres were injected into the left atrium. After 90 minutes of reperfusion, the heart was removed and sliced transversely. Areas not perfused by microspheres (areas at risk) were traced, planimetered and compared with the areas of necrosis after incubation in triphenyltetrazolium chloride. Areas were then converted into weights. In control dogs, the weight of necrotic myocardium was not significantly different from the weight at risk (5.0 +/- 0.7 vs 7.0 +/- 0.8 g, respectively [mean +/- standard error of the mean]), whereas it was markedly reduced in treated dogs (5.9 +/- 0.5 vs 9.4 +/- 0.7 g, respectively, p less than 0.001). The weight of salvaged myocardium was 3.4 +/- 0.5 g in treated dogs vs 1.9 +/- 0.4 g in the control group (p less than 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)

Genetic diversity and phylogenetic analysis of human immunodeficiency virus type 1 subtypes circulating in French Guiana.

We investigated the characterization of different HIV-1 subtypes present in French Guiana by use of three different methods. Serological methods were used for the initial screening, which were then confirmed by the heteroduplex mobility assay (HMA). The V3 env region was subsequently sequenced for phylogenetic analysis, to confirm the subtype of the samples, and to assign a subtype to samples that gave results that were difficult to interpret or discordant by serology or HMA. A total of 221 HIV-1 seropositive samples were typed; 110 of them were confirmed by HMA and 16 were sequenced. Of the 221 samples tested 210 patients (95%) were found to be infected with subtype B, 10 (4.5%) were infected with subtype A, and one patient was infected with subtype F. Phylogenetic analysis demonstrated that the strains from French Guiana were closely related to the subtype A and B subtypes, and that one strain was closely related to an F subtype (100% bootstrap value). Four strains from French Guiana clustered in the subtype A (99% bootstrap value) and the other strains were associated with subtype B (100% bootstrap value). The geographic position of French Guiana suggested that HIV-1 was probably introduced into the country via several routes, and thus the pattern of the HIV-1 epidemic might evolve in the near future.

Primary B-cell gastric lymphomas of mucosa-associated lymphoid tissue. Histological and immunohistochemical study of ten cases on surgical specimens.

Ten cases of gastric mucosa-associated lymphoid-tissue B-cell lymphoma were studied on surgical specimens by histology and immunohistochemistry, with monoclonal and polyclonal antibodies for B- and T-cells. For the first time, percentage of centroblast-like cells was appreciated, using their LN1 positivity, by opposition to the negativity of centrocyte-like cells (LN2 immunoreactivity alone). Lymphomas were divided into four main groups: A) centrocyte-like cells; B) centrocyte-like cells and immunocytoma; C) centrocyte-like cells admixed to centroblast-like cells; and D) centrocyte-like cells, immunocytoma and at least 30% of centroblast-like cells. Group C was divided into 3 subgroups: C1 (rare centroblast-like cells); C2 (30-50% of centroblast-like cells); and C3 (predominant centroblast-like cells). Therefore low grade (A, B, C1) and high grade malignancy groups (C2, C3, D) were identified: this preliminary subdivision could be extended in larger series and applied to mucosa-associated lymphoid-tissue lymphoma from other sites. Furthermore, the possible prognostic significance of this subdivision could be evaluated by correlation with long term follow-up.

Peripheral T-cell lymphoma or Hodgkin's disease in a HIV seropositive patient: a histopathological study.

Malignant lymphomas occurring in patients with AIDS are usually derived from the B-cell lineage while T-cell malignant lymphomas are very rare in these patients. We report a HIV seropositive 29-year-old homosexual man in whom cervical lymph node biopsy showed an atypical lymphoproliferative process. On morphological and paraffin section immunohistochemical grounds the possibility of Hodgkin's disease (HD) mixed cellularity was initially suggested, but frozen section immunohistochemical studies revealed that the cellular infiltrate exhibited an aberrant pan T immunophenotype and consequently the diagnosis of peripheral T-malignant lymphomas (T-ML) was made. However, genotypic studies would be required to definitely confirm this diagnosis, in such cases. In our case, varying numbers of small and medium-sized cells were positive for both Leu 3/CD4 and Leu 2/CD8 whereas some large cells reacted only with Leu 3/CD4 antibody. Some medium-sized, large and giant cells showed cytoplasmic positivity for Leu M1/CD15. Furthermore, the positivity of many large and giant cells with the activation markers BerH2/CD30, Ki-1/CD30, Tac/CD25 and HLA-DR suggested an activation state for these cells. Our findings emphasize the usefulness of frozen section immunohistochemical methods in order to investigate the spectrum of lymphoid malignancies occurring in HIV seropositive patients, and confirm results of previous studies which stressed the diagnostic difficulties that may appear in distinguishing HD from peripheral T-ML.

TH mRNA over-expression in rats with chronic excitotoxic striatal lesions.

Twenty weeks after ibotenic acid lesions of the striatum, the amount of tyrosine hydroxylase (TH) in this structure was markedly increased. This was accompanied by a 3-fold increase in TH mRNA levels in the ipsilateral subtantia nigra (SN). Striatal levels of dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) were markedly reduced. In the nucleus accumbens, spared by the lesion, DA neurotransmission was also altered, as evidenced by a reduction of DA and DOPAC, but no increase in TH could be detected. TH mRNA levels were moderately enhanced in the ventral tegmental area (VTA). Thus, lesioning in the striatum induces TH gene activation in both SN and VTA neurones, not strictly related to DA function at the terminal level.

Retrograde coronary sinus perfusion: a safe alternative for ensuring cardioplegic delivery in aortic valve surgery.

During aortic valve surgery, cardioplegic solution is delivered through direct cannulation of both coronary ostia. Since this approach may cause an intimal injury leading to acute dissection or late ostial stenosis, we have evaluated retrograde coronary sinus perfusion (RCSP) as a means of delivering cardioplegia in 12 patients undergoing aortic valve replacement. The retroperfusion of the cardioplegic solution was performed with a balloon-tipped catheter inserted into the coronary sinus through the right atrium. The perfusion pressure averaged 40 mm Hg. Twelve patients undergoing antegrade coronary perfusion served as controls. Both groups were matched for preoperative and intraoperative data. The postoperative evaluation focused on hemodynamic status, as evidenced by serial measurements of right-sided pressures and cardiac output at 1, 6, 12, 18, and 24 hours after operation. The stroke volume index and the left ventricular and right ventricular systolic stroke work indexes were then calculated. There was no statistically significant difference between the two groups. We conclude that RCSP is a simple, safe, and effective means of cardioplegic protection during aortic valve surgery.

Regulation of tyrosine hydroxylase gene expression in mesencephalic dopamine neurons: effect of imipramine treatment.

The effects of a chronic imipramine treatment on the mesoamygdaloid pathway of rats were examined. Using semiquantitative immunocytochemical techniques, it was observed that the level of TH mRNA was decreased in the ventral tegmental area (VTA). In contrast, the TH protein was increased in both the VTA and amygdala. The TH activity was decreased in the amygdala when assessed under normal conditions but increased after a preincubation to phosphorylate the enzyme, suggesting a lowering of the protein-specific activity in the terminals. These results show that TH protein turnover in the mesoamygdaloid neurons can be reduced by chronic imipramine treatments, thereby producing an accumulation of inactive TH protein in the neurons while also decreasing TH gene activity in the cell bodies.

Polydioxanone in digestive surgery. An experimental study.

An experimental study was carried out to analyze the reaction of polydioxanone suture. Two hundred ten single layer colonic anastomoses were performed in rats and histopathologically studied at nine different times between the 2nd and 180th postoperative days. Eighteen of 105 abdominal wound closures were studied in the same way between the 7th and 90th days. Polydioxanone was resorbed slowly in approximately 6 months with minimum inflammation. We believe that polydioxanone is actually the suture material of choice for all biliary and biliodigestive sutures.

Histological, immunohistochemical, ultrastructural and biochemical study of human gastric composite tumor: expression of the serotonin-2B receptor by the neuroendocrine component.

We report a case of a human gastric composite tumor occurring seven years after a partial gastrectomy for a low grade B cell MALT lymphoma. Histological examination of the tumor revealed two intimately intermingled components: 1. A moderately to poorly differentiated tubulo-acinar adenocarcinoma with signet-ring cells; and 2. Isolated or clustered small neuroendocrine cells without atypia expressing chromogranin A, somatostatin and/or glucagon, serotonin (5-HT) and, the 5-HT2B receptors. In addition to immunohistochemical detection, the presence of 5-HT2B receptors was shown pharmacologically through [125I]-DOI binding. Since 5-HT2B receptors have been demonstrated to have autocrine functions and, mitogenic and transforming properties, these results suggest a role of 5-HT in neuroendocrine malignant transformation. On the other hand, the expression of somatostatin and the detection by reverse transcriptase polymerase chain reaction (RT-PCR) of somatostatin receptor subtypes 2, 3, and 5, which have been shown to be involved in tumor regression, might account for the long evolution of this case (> 5 yr). This case illustrates the importance of local humoral modulation in tumor growth. Moreover, ultrastructural results favor a unique origin of the tumor cells from one amphicrine cell type.

Nucleolar organizer regions (NORs) staining and proliferating cell nuclear antigen (PCNA) immunostaining in mucosa-associated lymphoid tissue (MALT) gastric lymphomas.

Nucleolar organizer region-associated proteins (AgNOR) and proliferating cell nuclear antigen (PCNA) have been studied by means of a silver staining technique and immunohistochemistry, in paraffin-embedded, gastrectomy specimens of 12 low-grade and 13 high-grade gastric MALT lymphomas respectively. A significant difference was found between the AgNOR count and PCNA index of low-grade lymphomas (mean AgNOR count 2.5 and mean PCNA index 8.33%) and high-grade lymphomas (mean AgNOR count 8.67 and mean PCNA index 49.7%). It is suggested that both methods are useful adjuncts to histopathology for the distinction between low and high grade gastric MALT lymphomas. We also found heterogeneity in AgNOR counts and PCNA index among individual cases of either low or high grade MALT gastric lymphomas. This suggests that the AgNOR count and PCNA index is helpful in the individual approach of the proliferation rate of each tumour, a parameter of potential importance for predicting the biological behaviour of the tumour and the prognosis of the disease.

Steel pressure vessels for hydrostatic pressures to 50 kilobars.

Cylindrical steel pressure vessels are described that can be used for hydrostatic pressures up to 50 kilobars. Monoblock vessels of 350 maraging steel can be used to 40 kilobars and compound vessels with an inner vessel of 350 maraging steel and an outer vessel of 300 maraging steel to 50 kilobars. Neither requires the cylinder to be end loaded, and so they are much easier to use than the more usual compound vessels with a tungsten carbide inner and steel outer vessel.

Glass windows for hydrostatic pressures of 45 kbar.

Unprotected glass windows can be used at hydrostatic pressures up to at least 45 kbar on a 0.5-mm-diam aperture, and up to 40 kbar on a 0.8-mm-diam aperture.

Effect of peroperative hypertension on 125I-albumin uptake by vein graft media.

Aortocoronary bypass, in use for over 20 years, is followed by lesions in 80% of vein grafts after 10 years. In 8 patients (mean age 60, SD = 7.8) before cardiopulmonary bypass, segments of saphenous vein were harvested, and following hydraulic distension (760 mmHg x 10 min), stripped of adventitia, de-endothelialized, cannulated, immersed in oxygenated Krebs solution pH 7.4 37 deg C, and pressurized (100 mmHg) with 4% 125I-albumin-Evans blue for two hours. The samples were then frozen, and serially sectioned in the plane of the lumenal surface. The radioactivity of the 20 microns-thick sections was then determined, and expressed as a tissue: labelled solution concentration ratio (cpm unit volume wet tissue/cpm unit volume labelled solution). Distended portions had a higher mean transmural concentration ratio, as compared with the undistended portion; the ratio distended/undistended was 1.23, SD = 0.17, n = 8, p less than 0.01 (paired t test). This suggests alteration in the transport properties of vein interstitium by hypertension. Random histologic examination of native veins revealed no structural damage apart from age-related dystrophy of elastic fibres and intimal and medial fibrosis in grossly normal veins. These results suggest that the quality of the vein used, combined with hypertension during its preparation, may be factors in the genesis of postoperative graft failure.