Ultrastructural aspects of virus replication in one fatal case and several other isolates from a dengue type 2 outbreak in Rio de Janeiro

Dengue virus replication in mosquito cell cultures was observed by electron microscopy in one fatal and 40 classical isolates from a dengue type 2 outbreak in Rio de Janeiro and compared with the prototype New Guinea C strain. All the Brazilian isolates presented, beside the classical structured dengue virus particles, fuzzy coated virus-like particles, never observed in thereferencial New Guinea C virus strain. more numerous DEN-2 virus particles, fuzzy coated virus-like particles, defective virus particles and smooth membrane structures inside the rough endoplasmic reticulum characterized the unique fatal isolate examined.

Els horts urbans a la ciutat de Barcelona: les experiències d'horts urbans comunitaris com a formes d'intervenció social i ambiental

Aquest estudi presenta la situació actual dels horts urbans (i periurbans) a la ciutat de Barcelona, els quals s'han classificat segons el tipus d'organització desenvolupada a cada projecte. Així, podem trobar horts de gestió: a) individual i autogestionada; b) comunitària i autogestionada; c) individual i supervisada, i d) comunitària i supervisada. Els horts urbans es presenten, en general, com una eina interessant en la millora de la sostenibilitat urbana. A més de tenir una clara funció d'entreteniment, són propostes que consideren la internalització a les ciutats de la producció de part dels aliments que s'hi consumeixen i alhora aprofiten part dels residus que s'hi produeixen. En particular, els horts urbans comunitaris i autogestionats – el centre d'aquest estudi – es plantegen com espais de participació i autogestió d'acord a la complexitat del context local, d'integració social a través de noves formes de relació i de creació, d'educació ambiental i de transmissió i intercanvi de coneixements inter-generacional. A més, es presenten com una alternativa d'organització realment participativa del territori urbà. Finalment, i d'acord amb l'anterior, es destaquen un conjunt de característiques dels projectes d'horts urbans comunitaris, que juguen un rol fonamental en la capacitat d'aquests per intervenir en aspectes socials i ambientals de la ciutat; característiques que s'haurien de tenir en compte a l'hora de promocionar i implementar projectes d'horts urbans de qualsevol tipus.

The L-Type Ca+ and KATP channels may contribute to pacing-induced protection against anoxia-reoxygenation in the embryonic heart model.

L-Type Ca(2+) and K(ATP) Channels in Pacing-Induced Cardioprotection. AIMS: The L-type Ca(2+) channel, the sarcolemmal (sarcK(ATP)), and mitochondrial K(ATP) (mitoK(ATP)) channels are involved in myocardial preconditioning. We aimed at determining to what extent these channels can also participate in pacing-induced cardioprotection. METHODS: Hearts of 4-day-old chick embryos were paced in ovo during 12 hour using asynchronous intermittent ventricular stimulation at 110% of the intrinsic rate. Sham operated and paced hearts were then submitted in vitro to anoxia (30 minutes) and reoxygenation (60 minutes). These hearts were exposed to L-type Ca(2+) channel agonist Bay-K-8644 (BAY-K) or blocker verapamil, nonselective K(ATP) channel antagonist glibenclamide (GLIB), mitoK(ATP) channel agonist diazoxide (DIAZO), or antagonist 5-hydroxydecanoate. Electrocardiogram, electromechanical delay (EMD) reflecting excitation-contraction (E-C) coupling, and contractility were determined. RESULTS: Under normoxia, heart rate, QT duration, conduction, EMD, and ventricular shortening were similar in sham and paced hearts. During reoxygenation, arrhythmias ceased earlier and ventricular EMD recovered faster in paced hearts than in sham hearts. In sham hearts, BAY-K (but not verapamil), DIAZO (but not 5-hydroxydecanoate) or GLIB accelerated recovery of ventricular EMD, reproducing the pacing-induced protection. By contrast, none of these agents further ameliorated recovery of the paced hearts. CONCLUSION: The protective effect of chronic asynchronous pacing at near physiological rate on ventricular E-C coupling appears to be associated with subtle activation of L-type Ca(2+) channel, inhibition of sarcK(ATP) channel, and/or opening of mitoK(ATP) channel.

Wyeomyia luteoventralis Theobald, the type species of the subgenus Dendromyia Theobald (Diptera: Culicidae)

Wyeomyia (Dendromyia) luteoventralis is redescribed from females, males, larvae and pupae collected at the type locality, in Brazil, and compared to the closely related species. At least six species - Wy. luteoventralis, Wy. ypsipola, Wy. testei, Wy. trifurcata, Wy. complosa and Wy. jocosa - appparently, belong to a monophyletic group, the subgenus Dendromyia. All other species previously included in Dendromyia are hereby excluded from this subgenus and retained in genus Wyeomyia without subgeneric placement.

Potassium channel alterations mediate peripheral nerve hyperexcitability in a mouse model of type 2 diabetes mellitus

Diabetes mellitus (DM) is a major cause of peripheral neuropathy. More than 220 million people worldwide suffer from type 2 DM, which will, in approximately half of them, lead to the development of diabetic peripheral neuropathy. While of significant medical importance, the pathophysiological changes present in DPN are still poorly understood. To get more insight into DPN associated with type 2 DM, we decided to use the rodent model of this form of diabetes, the db/db mice. During the in-vivo conduction velocity studies on these animals, we observed the presence of multiple spiking followed by a single stimulation. This prompted us to evaluate the excitability properties of db/db peripheral nerves. Ex-vivo electrophysiological evaluation revealed a significant increase in the excitability of db/db sciatic nerves. While the shape and kinetics of the compound action potential of db/db nerves were the same as for control nerves, we observed an increase in the after-hyperpolarization phase (AHP) under diabetic conditions. Using pharmacological inhibitors we demonstrated that both the peripheral nerve hyperexcitability (PNH) and the increased AHP were mostly mediated by the decreased activity of Kv1-channels. Importantly, we corroborated these data at the molecular level. We observed a strong reduction of Kv1.2 channel presence in the juxtaparanodal regions of teased fibers in db/db mice as compared to control mice. Quantification of the amount of both Kv1.2 isoforms in DRG neurons and in the endoneurial compartment of peripheral nerve by Western blotting revealed that less mature Kv1.2 was integrated into the axonal membranes at the juxtaparanodes. Our observation that peripheral nerve hyperexcitability present in db/db mice is at least in part a consequence of changes in potassium channel distribution suggests that the same mechanism also mediates PNH in diabetic patients. ∗Current address: Department of Physiology, UCSF, San Francisco, CA, USA.

Individual contributions of mutant protease and reverse transcriptase to viral infectivity, replication, and protein maturation of antiretroviral drug-resistant human immunodeficiency virus type 1.

Human immunodeficiency virus type 1 (HIV-1) variants resistant to protease (PR) and reverse transcriptase (RT) inhibitors may display impaired infectivity and replication capacity. The individual contributions of mutated HIV-1 PR and RT to infectivity, replication, RT activity, and protein maturation (herein referred to as "fitness") in recombinant viruses were investigated by separately cloning PR, RT, and PR-RT cassettes from drug-resistant mutant viral isolates into the wild-type NL4-3 background. Both mutant PR and RT contributed to measurable deficits in fitness of viral constructs. In peripheral blood mononuclear cells, replication rates (means +/- standard deviations) of RT recombinants were 72.5% +/- 27.3% and replication rates of PR recombinants were 60.5% +/- 33.6% of the rates of NL4-3. PR mutant deficits were enhanced in CEM T cells, with relative replication rates of PR recombinants decreasing to 15.8% +/- 23.5% of NL4-3 replication rates. Cloning of the cognate RT improved fitness of some PR mutant clones. For a multidrug-resistant virus transmitted through sexual contact, RT constructs displayed a marked infectivity and replication deficit and diminished packaging of Pol proteins (RT content in virions diminished by 56.3% +/- 10.7%, and integrase content diminished by 23.3% +/- 18.4%), a novel mechanism for a decreased-fitness phenotype. Despite the identified impairment of recombinant clones, fitness of two of the three drug-resistant isolates was comparable to that of wild-type, susceptible viruses, suggestive of extensive compensation by genomic regions away from PR and RT. Only limited reversion of mutated positions to wild-type amino acids was observed for the native isolates over 100 viral replication cycles in the absence of drug selective pressure. These data underscore the complex relationship between PR and RT adaptive changes and viral evolution in antiretroviral drug-resistant HIV-1.

Dried blood spots collected on filter paper: an international resource for the diagnosis and genetic characterization of human immunodeficiency virus Type-1

The collection of dried blood spots (DBS) on filter paper provides a powerful approach for the development of large-scale, population-based screening programs. DBS methods are particularly valuable in developing countries and isolated rural regions where resources are limited. Large numbers of field specimens can be economically collected and shipped to centralized reference laboratories for genetic and (or) serological analysis. Alternatively, the dried blood can be stored and used as an archival resource to rapidly establish the frequency and distribution of newly recognized mutations, confirm patient identity or track the origins and emergence of newly identified pathogens. In this report, we describe how PCR-based technologies are beginning to interface with international screening programmes for the diagnosis and genetic characterization of human immunodeficiency virus type 1 (HIV-1). In particular, we review recent progress using DBS specimens to resolve the HIV-1 infection status of neonates, monitor the genetic evolution of HIV-1 during early infancy and establish a sentinel surveillance system for the systematic monitoring of HIV-1 genetic variation in Asia.

L'ús dels clítics pronominals del català i la seva adquisició per parlants de romanès i de tagal

El nostre estudi ha analitzat els mecanismes de transferència de la L1 en dos grups de parlants, un que té el romanès i l’altre el tagal com a llengües mare, en el procés d’aprenentatge del català com a L2. En concret ens hem centrat en l’estudi de l’ús dels clítics pronominals, una de les parts de la gramàtica catalana més apassionant per les seves idiosincràsies i la seva diversitat formal. L’ús d’aquest tipus de pronoms és un aspecte que resulta complicat d’adquirir per parlants de llengües en què no existeixen o on l’ús que en fan és diferent. En el nostre estudi, ens hem basat inicialment en la descripció de les característiques fonamentals dels clítics pronominals del català, del romanès i del tagal, a partir de bibliografia especialitzada i més general, i, en el cas del català, fent servir també la pròpia competència. Després d’aquest apartat més descriptiu, hem utilitzat un corpus d’entrevistes fetes en català a dos grups de parlants, el primer amb aprenents de català que tenen el romanès com a L1 i el segon amb aprenents de català que tenen el tagal com a L1, per analitzar quantitativament i qualitativament l’ús dels clítics pronominals. Un altre grup d’entrevistes, aquesta vegada a persones autòctones (català L1), ens ha servit com a grup de control. El nostre estudi dóna suport a la hipòtesi que atorga una importància especial a la transferència de la L1 en l’adquisició d'una segona llengua en general, i en la dels clítics pronominals d’una llengua romànica, en particular. Els resultats mostren que hi ha diferències entre els dos grups d'aprenents que són estadísticament significatives (ús del programa SPSS) i poden ser atribuïdes a les característiques de la L1.

The type specimens of mosquitoes (Diptera, Culicidae) deposited in the entomological collection of the Instituto Oswaldo Cruz, Rio de Janeiro, Brazil

A list of type specimens of 100 mosquito species deposited in the Entomological collection of the Instituto Oswaldo Cruz is presented. It includes five holotypes belonging to the subfamily Anophelinae; 56 holotypes of Culicinae and two of Toxorhynchitinae. A lectotype is designated for Toxorhynchites fluminensis. The holotypes of six nominal species - Psorophora chiquitana, Psorophora circunflava, Psorophora melanota, Psorophora lanei, Wyeomyia brucei and Uranotaenia noctivaga - previously considered non existent or of unknown location were found in the collection.

IRF6 Screening of Syndromic and a priori Non-Syndromic Cleft Lip and Palate Patients: Identification of a New Type of Minor VWS Sign.

Van der Woude syndrome (VWS), caused by dominant IRF6 mutation, is the most common cleft syndrome. In 15% of the patients, lip pits are absent and the phenotype mimics isolated clefts. Therefore, we hypothesized that some of the families classified as having non-syndromic inherited cleft lip and palate could have an IRF6 mutation. We screened in total 170 patients with cleft lip with or without cleft palate (CL/P): 75 were syndromic and 95 were a priori part of multiplex non-syndromic families. A mutation was identified in 62.7 and 3.3% of the patients, respectively. In one of the 95 a priori non-syndromic families with an autosomal dominant inheritance (family B), new insights into the family history revealed the presence, at birth, of lower lip pits in two members and the diagnosis was revised as VWS. A novel lower lip sign was observed in one individual in this family. Interestingly, a similar lower lip sign was also observed in one individual from a 2nd family (family A). This consists of 2 nodules below the lower lip on the external side. In a 3rd multiplex family (family C), a de novo mutation was identified in an a priori non-syndromic CL/P patient. Re-examination after mutation screening revealed the presence of a tiny pit-looking lesion on the inner side of the lower lip leading to a revised diagnosis of VWS. On the basis of this data, we conclude that IRF6 should be screened when any doubt rises about the normality of the lower lip and also if a non-syndromic cleft lip patient (with or without cleft palate) has a family history suggestive of autosomal dominant inheritance.

The type of K-ras mutation determines prognosis in colorectal cancer.

BACKGROUND: Mutations involving the oncogene K-ras in colorectal cancer may be related to tumor aggressiveness. However, the value of K-ras gene determination as a prognostic marker has not been clearly established. PATIENTS AND METHODS: The results from 98 patients recruited in a prospective study analyzing the effect of a K-ras mutation as a prognostic factor in colorectal cancer are reported. RESULTS: Disease-free (P = 0.02) and overall survival (P = 0.03) were significantly reduced for patients harboring a K-ras mutation. Two specific mutations demonstrated a significantly increased risk of disease recurrence, namely, 12-TGT (P = 0.04) and 13-GAC substitutions (P = 0.002). Patients with either of these substitutions had a 2-year disease-free survival rate of 37% compared with that of 67% for the group of patients harboring any other mutation type or a wild-type status (P = 0.01). CONCLUSIONS: The results herein presented suggest that K-ras acts as a prognostic factor in colorectal cancer and that this effect is probably related to a limited number of defined mutations.