[Plan for Improving the Initial Ambulation of Acute Stroke Patients].

BACKGROUND & PROBLEMS: Acute stroke patients should receive a rehabilitation assessment within 24-48 hours of hospitalization. Initial ambulation is known to reduce the occurrence of complications, improve the ability to perform activities of daily living, and reduce the risk of long-term disability. PURPOSE: To raise the initial ambulation willingness of acute stroke patients and to increase the willingness of these patients to receive rehabilitation treatment as soon as possible in order to reduce the long-term physical damage of the stroke incident. RESOLUTIONS: To develop and implement standard operating procedures for the initiation of ambulation (first time leaving the hospital bed) in acute stroke patients, to use health education brochures with texts and illustrations, and to have nurses physically assist patients to initiate ambulation. RESULTS: The rate of ambulation initiation in acute stroke patients rose from 32.0% pre-intervention to 85.4% post-intervention. CONCLUSIONS: Acute stroke patients who initiate ambulation soon after experiencing a stroke may reduce their risk of acute complications, increase their ability to perform activities of daily living, and reduce the risk of long-term disability. Thus, encouraging early ambulation is extremely important to improving the prognosis of this patient population.

Hepatitis C virus NS5A protein enhances gluconeogenesis through upregulation of Akt-/JNK-PEPCK signalling pathways.

BACKGROUND & AIMS: Hepatitis C virus (HCV) infection is highly associated with the type 2 diabetes mellitus, but the detailed mechanisms by which the viral proteins are involved in the clinical outcome remain unclear. METHODS: A cDNA microarray analysis was performed following introducing an NS5A-encoding plasmid or a control vector into a mouse system by hydrodynamics- based transfection. Differentially expressed genes that are associated with gluconeogenesis were selected and their expression levels in HCV patients, in NS5A-expressing systems, and in the viral subgenomic replicon system were further examined by real-time quantitative polymerase chain reaction and Western blot analysis. RESULTS: Differential gene expression including an upregulation of the gluconeogenic rate-limiting enzyme phosphoenolpyruvate carboxykinase (PEPCK) compared with controls was detected in mouse hepatocytes expressing HCV NS5A and in HCV patients with diabetes. In addition, an NS5A-dependent increase in glucose production was demonstrated in human primary hepatocytes. The upregulation of PEPCK and peroxisome proliferator-activated receptor-c coactivator-1a (PGC-1a) were also detected in NS5A-expressing cells and in the viral genotype 1b subgenomic replicon system. Further studies demonstrated that the NS5A-mediated upregulation of PEPCK and PGC-1a genes were resulted from the activation of PI3K-Akt and JNK signalling pathways. In addition, the expression levels of the forkhead transcription factor FoxO1 and the liver-enriched transcription factor HNF-4a were increased in HCV NS5A expressing cells. CONCLUSIONS: By upregulating the expression of PEPCK gene via its transactivators FoxO1 and HNF-4a, and the coactivator PGC-1a, the NS5A promotes the production of hepatic glucose which may contribute to the development of HCV-associated type 2 diabetes mellitus.

Lower prevalence of tuberculosis infection in BCG vaccinees: a cross-sectional study in adult prison inmates.

OBJECTIVES: To address whether the effect of BCG vaccination against tuberculosis (TB) infection lasts to adulthood. METHODS: A cross-sectional study on the prevalence of latent TB infection (LTBI) among HIV-negative men, using QuantiFERON-TB Gold In-tube (QFT-IT), was conducted at a prison in northern Taiwan with >3000 inmates. A QFT-IT ≥0.35 IU/ml was defined as LTBI. A QFT-IT ≥0.7 IU/ml was defined as recent LTBI. The association between the number of BCG scars and LTBI stratified by age was analysed. The study procedure was approved by the institutional review board, and all participants gave written informed consent before receiving screening tests. RESULTS: Among the 2385 participants, 25% had a QFT-IT ≥0.35 IU/ml. Increasing LTBI (14%, 32% and 50%) was observed with increased age (18-34 years, 35-54 years and ≥55 years) (p<0.001 by the Cochran-Armitage Trend Test). The number of BCG scars were found to be inversely correlated with QFT-IT results for both LTBI and recent LTBI in all three age groups (p<0.001 by Cochran-Mantel-Haenszel statistics). CONCLUSIONS: Our results suggest that BCG vaccine seems to have a protective effect in adults decades after vaccination according to the number of recent infections (QFT-IT ≥0.7 IU/ml). This finding has important implications for national policy of BCG vaccination. Further prospective cohort studies on the protective effect of BCG vaccination against TB infection in adults are warranted.

Ivory identification by DNA profiling of cytochrome b gene.

Ivory can be visually identified in its native form as coming from an elephant species; however, determining from which of the three extant elephant species a section of ivory originates is more problematic. We report on a method that will identify and distinguish the protected and endangered elephant species, Elephas maximus or Loxodonta sp. To identify the species of elephant from ivory products, we developed three groups of nested PCR amplifications within the cytochrome b gene that generate amplification products using highly degraded DNA isolated from confiscated ivory samples dating from 1995. DNA from a total of 382 out of 453 ivory samples were successfully isolated and amplified leading to species identification. All sequences were searched against GenBank and found to match with E. maximus and Loxodonta sp. with at least 99% similarity. The samples that were tested came from eight Asian elephants, 14 African forest elephants (Loxodonta cyclotis), and 360 African savannah elephants (Loxodonta africana). This study demonstrates a high success rate in species identification of ivory by a nested PCR approach within the cytochrome b gene which provides the necessary information for the protection of endangered species conservation.

Species identification of Kachuga tecta using the cytochrome b gene.

A DNA technique has been established for the identification to species level of tortoises. The test on the shell of the animal was used to identify samples from the species Kachuga tecta. A total of 100 tortoise shell specimens collected from the National Council of Agriculture (COA), Taiwan, were used in this study. Primer pairs were designed to amplify partial DNA fragments of cytochrome b within the mitochondrial genome. The DNA data showed that among the 100 samples, there were four distinct haplotype DNA sequences, within which there were a total of 90 variable sites. Between haplotypes I and II, there was only 1 nucleotide difference at position 228. Between haplotypes I and III, 65 nucleotide differences were observed; haplotypes I and IV, 62 nucleotide differences; and haplotypes III and IV, 56 nucleotide differences were observed. There were 66 and 63 nucleotide differences between haplotypes II and III and haplotypes II and IV respectively. All four haplotypes were compared with the DNA sequences held at the GenBank and EMBL databases. The most similar species were K. tecta (haplotype I and II), Morenia ocellata (haplotype III) and Geoclemys hamiltonii (haplotype IV), and their respective mtDNA similarities were 99.5%, 99.3%, 89.9% and 99.5%. However, as haplotype III was only 89.9% homologous with M. ocellata, it would seem that this haplotype shows only a limited relationship with a similar species registered currently in these databases. The method established by this study is an additional method for the identification of samples protected under Convention International Trade in Endangered Species (CITES) and will improve the work for the preservation of the endangered species.

Species identification of rhinoceros horns using the cytochrome b gene.

Material suspected of originating from species of Rhinoceros is frequently seized by forensic organizations investigating trade in endangered species. At present identification of the species is possible by DNA sequencing of the material, such as powdered rhinoceros horns. The unambiguous identification of rhino products using a 402 bp fragment of cytochrome b gene was investigated. This DNA sequence may not only assist in the identification of the unknown sample, but can be used to determine the phylogenetic relationships of rhinoceros species. Sequences of suspect rhinoceros horns were compared with the sequences registered in GenBank. The maximum value of genetic distance among white rhinoceros was 0.0176, and 0.0333 among black rhinoceros. In the comparison among rhinoceros species, the greatest genetic distance was between black and Indian rhinoceros (0.1564). The rhinoceros sequences extracted from GenBank and 13 samples in this study were clustered and separated from other mammals. Holstein cow was used as an out-group and was clustered with cattle in the phylogenetic tree. The results of this phylogenetic study also showed that there were four major branches among rhinoceros species from a common origin. The amplification of the 402 bp fragment of the cytochrome b gene was found to be able to detect rhinoceros DNA even in the ratio of 1:19 with Holstein cow DNA. In the initial identification of species from unknown powdered material, all the unknown samples were found to be from rhinoceroses. In phylogenetic analysis, the results supported the morphological hypothesis. The method used in this study can be applied in the identification of processed products of rhinoceros horns, such as sculptures, daggers, powders or even mixture powdered prescriptions.

A highly polymorphic STR locus in Cannabis sativa.

We report on the first short tandem repeat (STR) locus to be isolated from the plant Cannabis sativa. The STR locus, isolated by a hybrid-capture enrichment procedure, was found to contain a simple sequence repeat motif of 6 bp. This 6 bp repeat motif showed no variation in repeat length but with minor variations in repeat unit sequences. The data show the locus to be highly polymorphic with the number of repeat units ranging from 3 to 40 in 108 screened samples. The observed heterozygosity was approximately 87.04%. The forward and reverse primers (CS1F and CS1R) produced no PCR products in cross-reaction study from 20 species of plants, including highly related species such as Humulus japonicus and Nicotiana tabacum. This hexanucleotide repeat DNA locus could be used to identify cannabis samples and predict their genetic relationship as the test is specific to C. sativa and is highly reproducible.