Probing mechanisms that underlie human neurodegenerative diseases in Drosophila.

The fruit fly, Drosophila melanogaster, is an excellent organism for the study of the genetic and molecular basis of metazoan development. Drosophila provides numerous tools and reagents to unravel the molecular and cellular functions of genes that cause human disease, and the past decade has witnessed a significant expansion of the study of neurodegenerative disease mechanisms in flies. Here we review the interplay between oxidative stress and neuronal toxicity. We cover some of the studies that show how proteasome degradation of protein aggregates, autophagy, mitophagy, and lysosomal function affect the quality control mechanisms required for neuronal survival. We discuss how forward genetic screens in flies have led to the isolation of a few loci that cause neurodegeneration, paving the way for large-scale systematic screens to identify such loci in flies as well as promoting gene discovery in humans.

A tool for diagnosis of Dicrocoelium dendriticum infection: hatching eggs and molecular identification of the miracidium.

DNA primers were designed from the 18S rRNA sequence from the relevant digenean trematode Dicrocoelium dendriticum to evaluate a polymerase chain reaction-based diagnostic method of this parasite from its eggs in faeces of naturally and experimentally infected sheep. In order to get DNA from D. dendriticum eggs, several hatching mechanisms were studied. Successful results were obtained when the eggs were frozen to -80 °C and/or in liquid nitrogen and then defrosted. This method allowed the opening of the egg operculum and the liberation of the miracidium. DNA from D. dendriticum adults and from hatching egg miracidia was obtained and an amplification single band of 1.95 kb was observed using primers designed for the total 18S rRNA sequence in both cases as well as when the template DNA was from adults of the closely related parasite Fasciola hepatica; in addition, a single and specific 0.8-kb band was obtained when primers based on an internal partial 18S rRNA sequence were used. The method showed to be useful not only in samples coming from adults, but in eggs from gall bladder and faeces as well. F. hepatica internal 18S rRNA primers were also designed and used as a negative control to prove that the eggs in faeces came from D. dendriticum and not from F. hepatica. A molecular tool able to detect a minimum of about 40 D. dendriticum eggs in one of the definitive host faeces has been developed for the first time and could provide a useful molecular tool to improve the conventional coprological diagnosis for detecting D. dendriticum eggs.

Two new Telebasis Selys, 1865 species (Odonata: Zygoptera: Coenagrionidae) from Western Colombia.

Telebasis Selys, 1865 is a genus well represented in Colombia, with 18 species distributed throughout the territory, one of them being endemic to the country. One of the most underexplored biogeographic regions of the country is the Choc, where we found the two new endemic species of Telebasis here described, Telebasis blasi sp. nov. and Telebasis noveloi sp. nov. In the description of both species, we provide detailed information about their distribution, diagnostic characteristics for males and females, pictures of living specimens and diagnostic characteristics, and a map showing the localities where each species was found. Finally, remarks on the conservation status of these species are presented; we consider that they are probably endangered, since they occupy small ranges and are threatened by several human activities.

On the evolution of mimicry in avian nestlings.

Batesian mimicry (BM), where a nontoxic species resembles a toxic species with aposematic coloring, has been recently described for a Neotropical species of the suboscine passerine (Laniocera hypopyrra). Understanding the order and series in which these characteristics evolved is unknown and requires character information from closely related taxa. Here, we trace the origin of mimetic traits and how they evolved by examining antipredator characteristics using images and other field-collected trait data from nest and nestlings along with data available in the literature for the Laniisominae clade and closely related taxa. We found that morphological modifications of the downy feathers appeared first in the broader clade leading to the Laniisominae clade followed by further morphological and behavioral characteristics within the Laniisominae clade leading to the full BM. Images of nestlings in the Laniisominae and closely related clades demonstrated the extent of antipredator and camouflage characteristics. We found a complex set of behavioral and morphological traits in this clade for reducing predation from hiding to camouflage to mimicry. We further propose the evolution of two distinctive mimicry strategies in the Laniisominae clade: (1) Batesian Mimicry, as described above and (2) Masquerade, resemblance to inedible objects commonly found in their local environment. This complex set of antipredator traits shed light on the diversity of antipredator characteristics in avian nestlings, particularly in neotropical areas where the avian diversity is highest. Unfortunately, there are hundreds of species in the neotropics that lack basic natural history information on nesting traits, and therefore, we are likely missing critical information on the diversity of antipredator characteristics across avian nestlings.

[Quality indicators in hip fracture care after the implementation of an orthogeriatrics team]. / Indicadores de calidad en atención a fractura de cadera tras la implementación de un equipo de ortogeriatría.

INTRODUCTION: The incidence of hip fracture will increase dramatically in the coming decades. It has been associated with high mortality and morbidity. Orthogeriatric programs have improved outcomes in fragility hip fracture worldwide. There is little evidence in Mexico on the impact of orthogeriatrics and adherence to quality indicators. OBJECTIVE: To compare adherence to quality indicators in hip fracture before and after the implementation of an orthogeriatrics team compared to the traditional model. MATERIAL AND METHODS: Quasi-experimental study conducted in León, Mexico. Adherence to quality indicators before and after an interconsultant orthogeriatrics team was compared. The indicators measured were: surgical delay, treatment for osteoporosis, early mobilization, type of implant and delirium. RESULTS: We included 83 patients. The surgical delay was reduced from 144 to 116 hours (28 hours), the percentage of patients with early surgery was increased from 7.9 to 18%, in addition the percentage of patients with treatment for osteoporosis was increased and the incidence of delirium was reduced. CONCLUSION: Adherence to indicators is low, however, it was possible to discreetly increase adherence to these indicators. Internal policies must be generated in public hospitals to improve this adherence and see its impact on long-term outcomes.

INTRODUCCIÓN: La incidencia de fractura de cadera se incrementará radicalmente en las siguientes décadas. Se ha asociado a una mortalidad y morbilidad elevada. Los programas de ortogeriatría en hospitales públicos han demostrado mejorar los resultados asistenciales. Hay poca evidencia en México sobre el impacto de la ortogeriatría y el apego a indicadores de calidad. OBJETIVO: Comparar el apego a indicadores de calidad en fractura de cadera antes y después de la implementación de un equipo de ortogeriatría comparado con el modelo tradicional. MATERIAL Y MÉTODOS: Estudio cuasiexperimental realizado en León, México. Se comparó la adherencia a indicadores de calidad antes y después de un equipo interconsultante de ortogeriatría. Los indicadores medidos fueron: demora quirúrgica, tratamiento para osteoporosis, movilización temprana, tipo de implante y delirium. RESULTADOS: Incluimos 83 pacientes. La demora quirúrgica se disminuyó de 144 a 116 horas (28 horas), se incrementó el porcentaje de pacientes con cirugía temprana de 7.9 a 18%, además se incrementó el porcentaje de pacientes con tratamiento para osteoporosis y se logró disminuir la incidencia de delirium. CONCLUSIÓN: El apego a indicadores es bajo; sin embargo, se logró incrementar discretamente el apego a estos indicadores. Se deben generar políticas internas en los hospitales públicos para mejorar esta adherencia y ver su impacto en los resultados a largo plazo.

Description of a new species of Andaeschna (Odonata: Aeshnidae) from the Western Colombian Andes.

Andaeschna is a small and poorly known genus of dragonflies that inhabits the Andes, from Venezuela to northwestern Argentina. Here we describe Andaeschna occidentalis sp. nov., first species of the genus recorded in the Western Andes, specifically from the Tatamá National Park in Colombia. Males of this species can be differentiated from the other four species in the genus by the unique shape of the distal segment of the vesica spermalis. Likewise, females can be distinguished by their smaller and broader cerci (slightly shorter than S9) and its subquadrate point. The discovery of this beautiful species expands the range of the genus, previously known only in the Eastern Andes.

Molecular identification of Nocardia species using the sodA gene: Identificación molecular de especies de Nocardia utilizando el gen sodA.

Currently for bacterial identification and classification the rrs gene encoding 16S rRNA is used as a reference method for the analysis of strains of the genus Nocardia. However, it does not have enough polymorphism to differentiate them at the species level. This fact makes it necessary to search for molecular targets that can provide better identification. The sodA gene (encoding the enzyme superoxide dismutase) has had good results in identifying species of other Actinomycetes. In this study the sodA gene is proposed for the identification and differentiation at the species level of the genus Nocardia. We used 41 type species of various collections; a 386 bp fragment of the sodA gene was amplified and sequenced, and a phylogenetic analysis was performed comparing the genes rrs (1171 bp), hsp65 (401 bp), secA1 (494 bp), gyrB (1195 bp) and rpoB (401 bp). The sequences were aligned using the Clustal X program. Evolutionary trees according to the neighbour-joining method were created with the programs Phylo_win and MEGA 6. The specific variability of the sodA genus of the genus Nocardia was analysed. A high phylogenetic resolution, significant genetic variability, and specificity and reliability were observed for the differentiation of the isolates at the species level. The polymorphism observed in the sodA gene sequence contains variable regions that allow the discrimination of closely related Nocardia species. The clear specificity, despite its small size, proves to be of great advantage for use in taxonomic studies and clinical diagnosis of the genus Nocardia.

Actualmente, para la identificación y clasificación bacteriana se utiliza como método de referencia la secuenciación el gen rrs que codifica al rRNA16S, en el caso del análisis de cepas del género Nocardia, sin embargo, no tiene el suficiente polimorfismo para diferenciarlas a nivel de especie lo que hace necesaria la búsqueda de blancos moleculares que puedan proporcionar una mejor identificación. El gen sodA (que codifica la enzima superóxido dismutasa) ha tenido buenos resultados en la identificación de especies de otros Actinomicetos. En este estudio se propone para la identificación y diferenciación a nivel de especie del género Nocardia. Se utilizaron 41 especies Tipo de diversas colecciones, se amplificó y secuenció un fragmento de 386 pb del gen sodA y se realizó un análisis filogenético comparando los genes rrs (1171 pb) hsp65(401pb) secA1 (494pb), gyrB (1195pb) y rpoB (401pb), las secuencias fueron alineadas utilizando el programa Clustal X, los árboles evolutivos de acuerdo con el método de "Neighbor-Joining"se hicieron con el programa Phylo_win y Mega 6. Se analizó la variabilidad específica del gen sodA del género Nocardia presentando una alta resolución filogenética, una variabilidad genética importante, especificidad y confiabilidad para la diferenciación de los aislados a nivel de especie. El polimorfismo observado en la secuencia del gen sodA contiene regiones variables que posibilitan la discriminación de especies de Nocardia estrechamente relacionadas, y una clara especificidad, a pesar de su pequeño tamaño, demostrando ser de gran ventaja para utilizarse en estudios taxonómicos y en el diagnóstico clínico del género Nocardia.

First case of isolation of Nocardia wallacei reported in Mexico.

Nocardia species are aerobic, Gram-positive bacteria with branched filaments reported as opportunistic microorganisms associated with infectious diseases of the skin. We report the isolation of N. wallacei in Mexico from a 43-year-old man, an HIV-positive construction worker who sought care for difficulty breathing and abundant sputum.

Biocheese: a food probiotic carrier.

This review describes some aspects related to the technological barriers encountered in the development and stability of probiotic cheeses. Aspects concerning the viability of probiotic cultures in this matrix are discussed and the potential of cheese as a biofunctional food carrier is analyzed, outlying some points related to health and safety. In general, the manufacture of probiotic cheese should have little change when compared with the elaboration of cheese in the traditional way. The physicochemical and technological parameters influencing the quality of these products have also to be measured so as to obtain a process optimization.

An SfiI restriction map of the Bacillus subtilis 168 genome.

A restriction map of 24 SfiI (GGCCN4/NGGCC) restriction fragments has been constructed for the Bacillus subtilis genome. The combined sizes of the fragments indicate a genome size of approx. 4.2 Mb. The SfiI fragments range in size from 7-730 kb. Genetic markers have been located on 19 of the SfiI fragments, and 69 genetic markers have been assigned to the SfiI restriction map.

Identification of some clinically significant actinomycetes.

Early recognition of infections caused by actinomycetes tend to be highly dependent on at least a tentative diagnosis derived from microbiological tests, since the clinical symptoms can be difficult to interpret. Reliable identification of clinically significant actinomycetes depends upon the application of taxonomic techniques that are not yet widely used in clinical laboratories. The value of rapid enzyme, chemical and molecular fingerprinting techniques is exemplified by their application to the identification of representatives of clinically significant actinomycete taxa.

Identification of Brucella abortus, B. canis, B. melitensis, and B. suis by carbon substrate assimilation tests.

By using the results of seven carbon substrate assimilation tests from the Biotype 100 system (bioMérieux, Marcy-l'Etoile, France), we correctly identified 79 (85.9%) of 92 Brucella strains tested. The specificity of the method varied from 97.4 to 100% depending on the species. Although a biological safety cabinet must be used, this method represents an easy and fast alternative for the identification of Brucella species.

Effects of arsenic exposure on the frequency of HPRT-mutant lymphocytes in a population of copper roasters in Antofagasta, Chile: a pilot study.

A pilot biomarker study was conducted to investigate the feasibility of using the hypoxanthine guanine phosphoribosyltransferase (HPRT) gene in peripheral blood lymphocytes as a biomarker for detecting genetic effects of arsenic exposure. Blood and urine samples were obtained from workers highly exposed to arsenic in a copper roasting plant in Antofagasta, Chile. Individuals were classified according to their job titles into three potential exposure groups: high, medium, and low. To confirm exposure, arsenic concentration was determined in urine samples. The HPRT mutant frequencies were measured in lymphocytes from 15 individuals ranging in age from 24 to 66 years. The mean mutant frequencies for the three exposure groups were: low (9 x 10(-6)), medium (11 x 10(-6)), and high (24 x 10(-6)). An increased mutant frequency was observed in the highly exposed group, but the response was so slight that it is not likely that this assay will be capable of providing dose-response information across a range of lower, more typical environmental arsenic levels.

Preliminary study on genetic variability of Dicrocoelium dendriticum determined by random amplified polymorphic DNA.

Genetic variability of adult specimens of Dicrocoelium dendriticum has been studied using random amplified polymorphic DNA (RAPD). The worms were collected from the infected livers of different sheep from several localities in León province (NW Spain). DNA fragments were amplified by means of decamer primer oligonucleotides of arbitrary sequence. Some primers produce complex and highly variable patterns of amplified DNA in D. dendriticum. Intra- and inter-population genetic variability of adult parasites were analyzed, scoring polymorphic and monomorphic reproducible bands by means of the Jaccard similarity, and dendrograms showing genetic relationships between individuals were obtained using the FITCH method. Genetic variability seems to be high in this parasite and genetic similarity within a population (worms infecting a single animal) is similar to the average similarity between worms from different sheep. These results suggest that each sheep is infected by numerous genetically different parasites from one or more populations of infected ants.

Estimation of the percolation thresholds in dextromethorphan hydrobromide matrices.

Percolation theory is a multidisciplinary theory that studies chaotic systems. It has been applied in the pharmaceutical field since 1987. Knowledge of the percolation threshold -- one of the most important concepts in percolation theory -- results in a clear improvement of the solid dosage form design. The percolation threshold is the concentration showing the maximum probability to obtain, for the first time, a percolating cluster of a substance. In this work, the percolation thresholds of dextromethorphan.HBr/Eudragit RS-PM inert matrices were estimated. The drug percolation threshold was estimated as 0.3691+/-0.0541 (P=0.05) of the total porosity (ranging between 23 and 36% w/w of drug). The SEM micrographs of the matrices are consistent with the estimated percolation range. In agreement with previous reports, different percolation thresholds were found for the matrix forming excipient Eudragit RS-PM. The site percolation threshold (based on the release properties) ranged between 10 and 20% v/v of the excipient, the site-bond percolation threshold (estimated from the mechanical properties) between 29.5 and 34% v/v of the excipient and the swelling percolation threshold between 34.3 and 46.9% v/v of the excipient. These percolation ranges are in agreement with those found previously for Eudragit RS-PM matrices containing naltrexone.HCl and morphine.HCl.

[Diagnosis and treatment of mycobacterial keratitis following LASIK. Case report and review of the literature]. / Diagnose und Therapie der Mykobakteriumkeratitis nach LASIK. Kasuistik und Literaturübersicht.

BACKGROUND: Mycobacterial keratitis is a rare complication following LASIK but can lead to an extremely unfavourable outcome. The diagnosis and treatment is often delayed due to confusion with other entities including diffuse lamellar keratitis and poor clinical outcomes with flap amputation and/or keratoplasty are often the case. PATIENT AND METHODS: We report the results of LASIK in a 51-year-old woman with subsequent early-diagnosed mycobacterial keratitis and compared this case to treatments and outcomes reported in the literature. RESULTS: The patient presented 10 days following LASIK with a white focal infiltrate in the stromal interface. The flap was lifted and cultures from the stromal bed and the reverse of the flap were obtained and the interface irrigated. The patient was treated with topical antibiotics (ciprofloxacin 0.3%, amikacin 2.5%, clarithromycin 40 mg/ml and tobramycin 15 mg/ml) for 8 weeks and at the most recent follow-up she had a visual acuity of 1.25. CONCLUSION: In a large number of published cases in the literature the flap had to be amputated and/or corneal transplants were necessary. Early diagnosis and treatment however, are essential to successfully treat post-LASIK keratitis. Therefore the patients should be followed up carefully in the early postoperative period.

[Diffuse lamellar keratitis. Postoperative prophylactic treatment with corticosteroids in an experimental animal study]. / Diffuse lamelläre Keratitis Tierexperimentelle Studie zur Evaluierung einer postoperativen Kortikosteroidprophylaxe.

PURPOSE: The aim of this experimental ani-mal study was to induce diffuse lamellar keratitis (DLK), and investigate a prophylactic treatment with corticosteroids. MATERIALS AND METHODS: A corneal flap was cut in 40 eyes from 20 Dutch-belted rabbits and the interface inoculated with either Pseudomonas aeruginosa lipopolysaccharide (LPS) endotoxin ( n=21) or Palmolive Ultra soap ( n=19). Half of the eyes were treated with topical corticosteroids and the other half remained untreated. Slitlamp examinations were performed 1, 3, 5 and 7 days postoperatively and DLK was graded from I-IV. RESULTS: At the end of the study 33 eyes were available for evaluation and 94% of the non-treated eyes developed DLK. Out of those eyes treated with steroids 19% developed DLK during the 1 week follow-up period. This was statistically significantly lower ( P=0.018) when compared to the untreated group. CONCLUSION: Pseudomonas aeruginosa LPS endotoxin as well as Palmolive((R)) Ultra caused a very high rate of DLK in rabbit eyes. The postoperative prophylactic treatment with corticosteroids showed a statistically significant lower DLK rate in this rabbit eye model.

[Diffuse lamellar keratitis: prophylactic treatment with ketorolac tromethamine 0.5% in an animal model]. / Queratitis lamelar difusa: tratamiento profiláctico con ketorolaco trometamina 0,5% en un modelo animal.

PURPOSE: The objective of this study is to evaluate the use of a topical non-steroidal anti-inflammatory drug in the treatment of induced diffuse lamellar keratitis (DLK) in an animal model. MATERIALS AND METHODS: A corneal flap was created in 40 eyes of 20 Dutch-belted rabbits using the ASC microkeratome. The interface was inoculated with either Pseudomonas Aeruginosa Lipopolysaccharide (LPS) endotoxin or Ultra Palmolive liquid dish washer. The rabbits were divided in two groups: Group I (n=20) treated with ketorolac tromethamine ophthalmic solution 0.5% 4 times a day and the group II (n=20) used as control. The rabbits were examined at the slit lamp at day 1, 3, 5 and 7 postoperatively. DLK was graded from I-IV. RESULTS: At the end of the study 31 eyes were available for evaluation. 28 eyes (90%) developed DLK: 86% of the treated group and 94% of the control group during the follow-up. The treated group showed a lower rate of DLK as well as a lower severity. However, no statistically significant difference was found when comparing both groups (P>0.05). CONCLUSION: Pseudomonas aeruginosa LPS endotoxin and Palmolive Ultra can induce DLK in rabbit eyes. The postoperative prophylactic treatment with a topical non-steroidal anti-inflammatory drug showed a tendency towards a lower DLK rate as well as the severity of the disease.

Experimental infection of Artibeus intermedius bats with serotype-2 dengue virus.

Dengue fever is caused by a flavivirus that primarily infects humans and Aedes sp. mosquitoes. However, viral replication in wild animals other than non-human primates has been scarcely studied. In this report, the susceptibility of Artibeus intermedius frugivorous bat to serotype-2 dengue virus (DENV-2) infection was tested. Twenty-three bats were intraperitoneally inoculated with different viral loads of DENV-2 (New Guinea-C strain). Forty-three percent of the infected bats developed bruises on the chest or on the wings. Histological analyses showed structural alterations in the spleen and bleeding in liver and intestine, but the virus was not detected by RT-PCR in any of the analyzed tissues, and it was found in only one bat (kidney) by semi-nested RT-PCR. In sera, the viral RNA was detected by semi-nested RT-PCR in 39% of bats, but only 8% of bats seroconverted. Overall, these data indicate that DENV-2 replicates poorly in these bats, suggesting they are not suitable hosts to this virus.