Detection of Mycoplasma species in cell culture by PCR and RFLP based method: effect of BM-cyclin to cure infections

PURPOSE: A two-stage nested polymerase chain reaction (PCR) assay system was described that amplifies the 16S-23S rRNA spacer region sequences of Mycoplasma and Acholeplasma infections in cell cultures and virus stocks. METHODS: Established cell lines and virus stocks were screened for the presence of Mycoplasma by using nested PCR using two sets of outer and inner primers, amplifies 16S-23S rRNA. PCR and restriction fragment length polymorphism (RFLP) assay was used to detect and identify most of the species-specific Mycoplasmas involved in cell cultures and virus stock contaminants. Infected cultures detected by PCR-RFLP were further treated with BM-cyclin (5 microg/mL) and passaged for three times and tested for Mycoplasma infections by PCR-RFLP. RESULTS: Mycoplasma pirum and Mycoplasma orale infections were detected by nested PCR. Species specificity was identified by using RFLP of Vsp I, Cla I and Hin dIII restriction enzymes. Mycoplasma infections were cured by treatment with BM-cyclin. This was further confirmed by non-amplification of PCR amplimers in BM-cyclin treated vs. non-treated cultures. CONCLUSIONS: Regular monitoring of cell cultures for Mycoplasma infections and identification of species-specific Mollicutes will identify the source of contaminations. This approach can be used for quality control of the biological reagents used in cell culture and virology laboratories.     

Duplex PCR Assay for Identification of Tissue of Cattle and Buffalo Origin Targeting Mitochondrial Cytochrome b Gene

Proceedings of the National Academy of Sciences, India Section B: Biological Sciences - The aim of this study was to develop PCR assay for simultaneous detection of tissues from cattle and buffalo...     

Consumer grievance reporting in G.M.C. Hospital, Miraj

A cross-sectional study conducted among 400 'consumers' at G.M.C. Hospital, Miraj revealed 91% consumers had one/more grievance/s. Higher grievances reported in illiterates, lower social class, indoor patients, surgical patients. Lower grievances reported in age < 15 yrs and 31-45 yrs, rural dwellers. Sex and new / old status of patients not affected grievance- reporting. 10 grievances out of 38 were found to be of serious nature. Some grievances (e.g. post-operative complications, discharge without clinical relief, absence of counselling, cursory clinical examination etc.) might have potential as possible medical negligence.     

Antipsychotic Potential and Safety Profile of TPGS-Based Mucoadhesive Aripiprazole Nanoemulsion: Development and Optimization for Nose-To-Brain Delivery

Delivering therapeutics to the brain using conventional dosage forms is always a challenge, thus the present study was aimed to formulate mucoadhesive nanoemulsion (MNE) of aripiprazole (ARP) for intranasal delivery to transport the drug directly to the brain. Therefore, a TPGS based ARP-MNE was formulated and optimized using the Box-Behnken statistical design. The improved in vitro release profile of the formulation was in agreement to enhanced ex vivo permeation through sheep mucous membranes with a maximum rate of permeation co-efficient (62.87  cm h^?1 × 10³) and flux (31.43  μg cm^?2.h^?1). The pharmacokinetic profile following single-dose administration showed the maximum concentration of drug in the brain (C_max) of 15.19 ± 2.51  μg mL^?1 and T_max of 1 h in animals with ARP-MNE as compared to 10.57 ± 1.88  μg mL^?1 and 1 h, and 2.52 ± 0.38  μg mL^?1 and 3 h upon intranasal and intravenous administration of ARP-NE, respectively. Further, higher values of % drug targeting efficiency (96.9%) and % drug targeting potential (89.73%) of ARP-MNE through intranasal administration were investigated. The studies in Wistar rats showed no existence of extrapyramidal symptoms through the catalepsy test and forelimb retraction results. No ex vivo ciliotoxicity on nasal mucosa reflects the safety of the components and delivery tool. Further, findings on locomotor activity and hind-limb retraction test in ARP-MNE treated animals established its antipsychotic efficacy. Thus, it can be inferred that the developed ARP-MNE could effectively be explored as brain delivery cargo in the effective treatment of schizophrenia without producing any toxic manifestation.     

Ambiguous Findings on Radiographs

Findings with uncertain clinical significance are frequently encountered on radiographs. A structure or opacity visible on radiographs could be due to several causes ranging from artifact or external structure to malignancy or a life-threatening process. The approach that a radiologist chooses to address ambiguous findings can have a significant impact on a patient's health. In this article we discuss the causes and impact of ambiguous findings on radiographs. We also discuss the various strategies radiologists can adopt to maximize clinical value and, when needed, reach a definite diagnosis.     

Changes in the range of motion of the cervical spine and adjacent segments at ≥24 months after uninstrumented corpectomy for cervical spondylotic myelopathy

Background  

Few clinical studies have described the changes in the range of motion (ROM) of the cervical spine and adjacent segments following central corpectomy. We aimed to quantify the changes in range of motion (ROM) of the cervical spine and the adjacent segments at ≥24 months following uninstrumented central corpectomy (CC) for cervical spondylotic myelopathy (CSM) and to determine the contribution of the adjacent segments to the compensation for loss of motion of the cervical spine following CC.