Design and synthesis of novel conformationally restricted HIV protease inhibitors

A set of HIV protease inhibitors represented by compound 2 has previously been described. Structural and conformational analysis of this compound suggested that conformational restriction of the P₁/P₂ portion of the molecule could lead to a novel set of potent protease inhibitors. Thus, probe compounds 3–7 were designed, synthesized, and found to be potent inhibitors of HIV protease.     

Antifeedant effects of in vitro culture extracts of the neem tree,Azadirachta indica against the desert locust (Schistocerca gregaria (Forskål))

Callus and micropropagated shoots were initiated from leaf explants of the neem tree, Azadirachta indica A. Juss. A variety of whole plant and in vitro cell cultures from neem seedlings of Ghanian origin were tested for insect antifeedant compounds using the desert locust (Schistocerca gregaria (Forskål)). Feeding suppression occurred when whole extracts of seed, leaf, callus, suspension and shoot cultures were tested in no-choice feeding bioassays. Controls of sucrose, carrot callus and the plant growth medium showed no feeding deterrence. Azadirachtin, the main known antifeedant in neem seed kernels, was quantified from a seed extract by HPLC but was not detected in any of the other extracts. Antifeedancy was determined during batch growth of a suspension culture which had been in culture for 5 months; results indicated that antifeedants were still being formed and that levels increased after maximum biomass was attained.     

Experimental Cross-Species Infection of Common Marmosets by Titi Monkey Adenovirus

Adenoviruses are DNA viruses that infect a number of vertebrate hosts and are associated with both sporadic and epidemic disease in humans. We previously identified a novel adenovirus, titi monkey adenovirus (TMAdV), as the cause of a fulminant pneumonia outbreak in a colony of titi monkeys (Callicebus cupreus) at a national primate center in 2009. Serological evidence of infection by TMAdV was also found in a human researcher at the facility and household family member, raising concerns for potential cross-species transmission of the virus. Here we present experimental evidence of cross-species TMAdV infection in common marmosets (Callithrix jacchus). Nasal inoculation of a cell cultured-adapted TMAdV strain into three marmosets produced an acute, mild respiratory illness characterized by low-grade fever, reduced activity, anorexia, and sneezing. An increase in virus-specific neutralization antibody titers accompanied the development of clinical signs. Although serially collected nasal swabs were positive for TMAdV for at least 8 days, all 3 infected marmosets spontaneously recovered by day 12 post-inoculation, and persistence of the virus in tissues could not be established. Thus, the pathogenesis of experimental inoculation of TMAdV in common marmosets resembled the mild, self-limiting respiratory infection typically seen in immunocompetent human hosts rather than the rapidly progressive, fatal pneumonia observed in 19 of 23 titi monkeys during the prior 2009 outbreak. These findings further establish the potential for adenovirus cross-species transmission and provide the basis for development of a monkey model useful for assessing the zoonotic potential of adenoviruses.     

Occurrence of Genes Associated with Enterotoxigenic and Enterohemorrhagic Escherichia coli in Agricultural Waste Lagoons

The prevalence among all Escherichia coli bacteria of the LTIIa toxin gene and STII toxin gene, both associated with enterotoxigenic E. coli, and of three genes (stxI, stxII, and eaeA) associated with enterohemorrhagic E. coli was determined in farm waste disposal systems seasonally for 1 year. Single- and nested-PCR results for the number of E. coli isolates carrying each toxin gene trait were compared with a five-replicate most-probable-number (MPN) method. The STII and LTIIa toxin genes were present continuously at all farms and downstream waters that were tested. Nested-MPN-PCR manifested sensitivity increased over that of single-MPN-PCR by a factor of 32 for LTIIa, 10 for STII, and 2 for the stxI, stxII, and eaeA genes. The geometric mean prevalence of each toxin gene within the E. coli community in waste disposal site waters after nested MPN-PCR was 1:8.5 E. coli isolates (1:8.5 E. coli) for the LTIIa toxin gene and 1:4 E. coli for the STII toxin gene. The geometric mean prevalence for the simultaneous occurrence of toxin genes stxI, stxII, and eaeA, was 1:182 E. coli. These findings based on total population analysis suggest that prevalence rates for these genes are higher than previously reported in studies based on surveys of single isolates. With a population-based approach, the frequency of each toxin gene at the corresponding disposal sites and the endemic nature of diseases on farms can be easily assessed, allowing farmers and public health officials to evaluate the risk of infection to animals or humans.     

Description of Pintomyia (Pifanomyia) paleotrichia, a miocene period new species from the Dominican Republic (Diptera: Psychodidae: Phlebotominae)

A new fossil species of phlebotomine sandflies is described from Dominican amber based in one specimen. Pintomyia (Pifanomyia) paleotrichia sp. nov. is distinguished from the other extant and extinct species by aspects of paramere and the basal tuft of bristles in the gonocoxite.     

Variation in xylem structure and function in roots and stems of scion–rootstock combinations of sweet cherry tree (<Emphasis Type="Italic">Prunus avium</Emphasis> L.)

To assess hydraulic architecture and limitations to water transport across scion–rootstock combinations (Prunus avium L. cultivar Van grafted on five differing size-controlling rootstocks: P. avium (vigorous) > CAB 11E > Maxma 14 > Gisela 5 > Edabriz (dwarfing)), we compared xylem anatomy, and calculated relative hydraulic conductivity (RC) and vulnerability index (VI) of roots (small, medium and large diameter) and stems. Water relations, leaf gas exchange and variations in growth were also determined. Roots exhibited larger-diameter xylem conduits (VD), greater RC and VI than stems in all Van–rootstock combinations. Moreover, there was a significantly higher vessel frequency (VF), lower VD, RC and VI in dwarfed trees, especially grafted on Gisela 5 than trees on the invigorating rootstocks, P. avium L., CAB 11E and Maxma 14. Anatomical constraints on water status imposed by the smaller VD (and/or in lower xylem thickness and root system length) of dwarfed trees imply a series of negative feedbacks, like a decrease in RC, stem water potential, leaf gas exchange and growth. On the other hand, Van grafted on CAB 11E and Maxma 14, with wide vessels and high VI, could be more susceptible to embolism, especially during periods of severe water stress, than trees on dwarfing rootstocks, which had small vessels and low VI.     

Photosensitization of wild and mutant strains of Escherichia coli by meso-tetra (N-methyl-4-pyridyl)porphine

Wild type Escherichia coli cells as well as some mutant strains lacking specific DNA repair systems are efficiently killed upon visible light-irradiation after 5 min-incubation with meso-tetra(4N-methyl-pyridyl)porphine (T4MPyP). The presence of oxygen is necessary for cell photoinactivation. The porphyrin appears to exert its phototoxic activity largely by impairing some enzymic and transport functions at the level of both the outer and cytoplasmic membrane. Thus, SDS-PAGE electrophoresis shows a gradual attenuation of some transport protein bands as the irradiation proceeds, while a complete loss of lactate and NADH dehydrogenase activities is caused by 15 min-exposure to light. On the other hand, DNA does not represent a critical target of T4MPyP photosensitization as suggested by the closely similar photosensitivity of the wild E. coli and E. coli strains defective for two different DNA repair mechanisms, as well as by the lack of any detectable alteration of the pUC19 plasmids extracted from photosensitized E. coli TG1 cells.     

Ca2+ changes the force sensitivity of the hair-cell transduction channel

The mechanically gated transduction channels of vertebrate hair cells tend to close in approximately 1 ms after their activation by hair bundle deflection. This fast adaptation is correlated with a quick negative movement of the bundle (a "twitch"), which can exert force and may mediate an active mechanical amplification of sound stimuli in hearing organs. We used an optical trap to deflect bullfrog hair bundles and to measure bundle movement while controlling Ca(2+) entry with a voltage clamp. The twitch elicited by repolarization of the cell varied with force applied to the bundle, going to zero where channels were all open or closed. The force dependence is quantitatively consistent with a model in which a Ca(2+)-bound channel requires approximately 3 pN more force to open, and rules out other models for the site of Ca(2+) action. In addition, we characterized a faster, voltage-dependent "flick", which requires intact tip links but not current through transduction channels.     

Diversity, distribution and abundance of sandflies (Diptera: Psychodidae) in Parana State, Southern Brazil

Due to the spread of Visceral leishmaniasis in Brazilian territory, this research was undertaken with the objective to identify the sandfly fauna and aspects of the species behavior in forest and anthropic environments in Paraná State, Brazil. The sandfly captures were sporadically carried out in 37 municipalities of the Paraná state, during the period from March 2004 to November 2005. Each municipality was once visited, when Falc?o traps were installed during three consecutive nights in forested fragments and anthropic environments; captures with Shannon traps installed in forests and edge of forests, inspection of walls and electric aspiration in domiciles and forested areas were sporadically undertaken. The data analysis was based on estimates of frequency and abundance of the species, by five native landscapes. A total of 38,662 specimens belonging to 23 species were captured. Nyssomyia neivai (Pinto) (75.6%), Ny. whitmani (Antunes & Coutinho) (10.1%), Migonemyia migonei (Fran?a) (7.8%), Expapillata firmatoi (Barreto et al.) (2.1%) and Pintomyia fischeri (Pinto) (1.6%) together constituted 97.2% of the phlebotomine collected. High frequencies of sand flies were found in forests, henhouses, pig-pens and domiciles. The three most frequent species have been implicated in transmission of tegumentary leishmaniasis in Paraná and in the Brazilian Southeastern and Southern Regions. Lutzomyia longipalpis (Lutz & Neiva), the visceral leishmaniasis vector, was not found, however Lutzomyia gaminariai (Cordero et al.), female of which is similar to that of Lu. longipalpis was captured, raising the need to investigate its behavior, including as regard to the vectorial competence.     

Molecular evidence for a recent demographic expansion in the puma (Puma concolor) (Mammalia,Felidae)

The puma is an iconic predator that ranges throughout the Americas, occupying diverse habitats. Previous phylogeographic analyses have revealed that it exhibits moderate levels of genetic structure across its range, with few of the classically recognized subspecies being supported as distinct demographic units. Moreover, most of the species’ molecular diversity was found to be in South America. To further investigate the phylogeographic structure and demographic history of pumas we analyzed mtDNA sequences from 186 individuals sampled throughout their range, with emphasis on South America. Our objectives were to refine the phylogeographic assessment within South America and to investigate the demographic history of pumas using a coalescent approach. Our results extend previous phylogeographic findings, reassessing the delimitation of historical population units in South America and demonstrating that this species experienced a considerable demographic expansion in the Holocene, ca. 8,000 years ago. Our analyses indicate that this expansion occurred in South America, prior to the hypothesized re-colonization of North America, which was therefore inferred to be even more recent. The estimated demographic history supports the interpretation that pumas suffered a severe demographic decline in the Late Pleistocene throughout their distribution, followed by population expansion and re-colonization of the range, initiating from South America.