Effective methods of preserving SCWL-diseased sugarcane leaves for genomic DNA extraction and molecular detection of phytoplasma

Simple and economical methods to preserve sugarcane leaf tissues infected with sugarcane white leaf (SCWL) phytoplasma for later detection of SCWL using nested- PCR analysis were described. Diseased leaf tissues were preserved by air-drying and stored at room temperature or stored fresh at 4 or -20°C in two types of bags: brown paper bags and plastic bags with punctured holes. Leaf materials were stored for up to 32 days prior to DNA extraction. The best storage condition was placing fresh leaves at -20°C and air-drying leaves in paper bags at room temperature. High molecular weight plant genomic DNA bands were detected in all stored infected tissues. Genomic DNAs were used as templates for nested- PCR which showed a single band of 210 bp in the 16S-23S intergenic spacer region specific to SCWL phytoplasma. Nested-PCR was also performed on genomic DNA extracted from diseased leaves heattreated in a microwave oven and the SCWL-specific bands were effectively detected. Therefore, treating diseased sugarcane leaves by air-drying, freezing, refrigeration or microwave heat treatment were all effective for molecular detection of SCWL phytoplasma. However, air-drying and storage in paper bags was the most economical and practical, especially when large numbers of samples are to be stored or transported from the fields to a distant laboratory or exchanged among laboratories.Key words: Sugarcane white leaf phytoplasma, nested-polymerase chain reaction (PCR), preservation, 16S- 23S intergenic spacer region

Illicit and Counterfeit Drug Analysis by Morphologically Directed Raman Spectroscopy

Morphologically directed Raman spectroscopy (MDRS) is a novel tool for the forensic analysis of illicit and counterfeit drug samples. MDRS combines Raman microspectroscopy with automated particle imaging so that physical and chemical information about the components of a mixture sample can be obtained. Results of automated particle imaging are used to determine samples for Raman analysis. The use of MDRS for these types of samples can be employed for both forensic investigations and adjudications of cases. The method provides insight about the physical and chemical composition of the sample, as well as about manufacturing and sample history. Here, MDRS was used in four different illicit and counterfeit drug analyses: (1) examination of a multicomponent drug mixture where the results could be used for comparative source attribution, (2) the detection of low (or trace) concentration particles in a drug sample, (3) the analysis of synthetic cathinone samples (i.e., bath salts), and (4) a study of counterfeit pharmaceutical products

Frailty syndrome in ambulatory patients with COPD

Panita Limpawattana,1 Siraphong Putraveephong,2 Pratchaya Inthasuwan,2 Watchara Boonsawat,3 Daris Theerakulpisut,4 Jarin Chindaprasirt5 1Division of Geriatric Medicine, 2Department of Internal Medicine, 3Division of Respiratory System, Department of Internal Medicine, 4Division of Nuclear Medicine, Department of Radiology, 5Division of Oncology Medicine, Department of Internal Medicine, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand Abstract: Frailty is a state of increased risk of unfavorable outcomes when exposed to stressors, and COPD is one of the several chronic illnesses associated with the condition. However, few studies have been conducted regarding the prevalence of COPD and its related factors in Southeast Asia. The objectives of this study were to determine the prevalence of frailty in COPD patients and to identify the associated factors in these populations. A cross-sectional study of COPD patients who attended a COPD clinic was conducted from May 2015 to December 2016. Baseline characteristics were collected, and the diagnosis of frailty was based on the FRAIL (fatigue, resistance, ambulation, illnesses, and loss of weight) scale. Descriptive statistics were used to analyze baseline data. Factors associated with frailty were analyzed using univariate and multivariate regression analyses. The results showed that the prevalence rates of frailty and pre-frailty were 6.6% (eight out of 121 cases) and 41.3% (50 out of 121 cases), respectively, among COPD patients. Fatigue was the most common component of the FRAIL scale that was found more frequently in frail patients than in non-frail patients (odds ratio [OR] 91.9). Factors associated with frailty according to multivariate analyses were comorbid cancer (adjusted OR [AOR] 45.8), at least two instances of nonelective admission over the past 12 months (AOR 112.5), high waist circumference (WC) (AOR 1.3), and presence of sarcopenia (AOR 29.5). In conclusion, frailty affected 6.6% of stable COPD patients. Cancer, two or more instances of nonelective hospitalization over the past 12 months, high WC, and presence of sarcopenia were associated with frailty. Early identification and intervention in high-risk patients is recommended to prevent or delay the adverse outcomes of frailty. Keywords: frailty syndrome, FRAIL scale, chronic lung disease, sarcopeni

Isolation of a gene preferentially expressed in mature anthers of rice (Oryza sativa L.)

Using monoclonal antibodies raised against pollen-specific proteins, we have isolated a cDNA clone, designatedOry-Cl from a rice anther cDNA expression library. A transcript corresponding to theOry-Cl gene showed preferential expression in anthers. This transcript was not detected in any vegetative tissues analysed. RNA gel blot analysis of different developmental stages of anthers showed that theOry-Cl gene is expressed at later stages of pollen development. In situ hybridisation showed that theOry-Cl transcript is only present in mature pollen.<br /