Establishment of loss-of-function phenotypes is often a key step in determining the biological function of a gene. We describe a procedure to obtain mutant petunia plants in which a specific gene with known sequence is inactivated by the transposable element dTphl. Leaves are collected from batches of 1000 plants with highly active dTphl elements, pooled according to a three-dimensional matrix, and screened by PCR using a transposon-and a gene-specific primer. In this way individual plants with a dTphl insertion can be identified by analysis of about 30 PCRs. We found insertion alleles for various genes at a frequency of about 1 in 1000 plants. The plant population can be preserved by selfing all the plants, so that it can be screened for insertions in many genes over a prolonged period.offspring and found to be due to insertions of dTphl elements (E.S., A.v.H., D.K., J.N.M.M., and R.K., unpublished data). Thus, the high incidence of mutations in the W138 line is mainly due to transposition of dTphl elements.We show how plants with a dTphl insertion in a specific gene can be identified in a large plant population by a polymerase chain reaction (PCR) assay. Such plants are heterozygous for the insertion allele unless the insertion occurred in a previous generation. After selfing, progeny homozygous for the insertion allele are obtained that can be analyzed for an altered phenotype. The screened plant population is maintained by selfing all plants, so that semipermanent libraries are obtained that can be screened at any time. These libraries are open to the scientific community.

In this study a combination of autologous chondrocyte implantation (ACI) and the osteochondral autograft transfer system (OATS) was used and evaluated as a treatment option for the repair of large areas of degenerative articular cartilage. We present the results at three years post-operatively. Osteochondral cores were used to restore the contour of articular cartilage in 13 patients with large lesions of the lateral femoral condyle (n = 5), medial femoral condyle (n = 7) and patella (n = 1). Autologous cultured chondrocytes were injected underneath a periosteal patch covering the cores. After one year, the patients had a significant improvement in their symptoms and after three years this level of improvement was maintained in ten of the 13 patients. Arthroscopic examination revealed that the osteochondral cores became well integrated with the surrounding cartilage. We conclude that the hybrid ACI/OATS technique provides a promising surgical approach for the treatment of patients with large degenerative osteochondral defects.

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Purpose To determine by B-scan ultrasonography if the addition of hyaluronidase affects the dispersal of anaesthetic fluid after sub-Tenon's injection. Design Single-centre prospective randomised double-blind study. Materials and methods We performed a trial in 19 patients who were randomised to receive 5 ml of lidocaine 2% alone, or with hyaluronidase 15 IU/ml. A pre-anaesthetic B-scan ultrasound was performed followed by a standard infero-nasal sub-Tenon's injection. Further B-scan ultrasound studies were performed at 1, 3, and 5 min recording depth of local anaesthetic fluid. Data was analysed with Fisher's exact test and Student's t-test where appropriate. Results were considered significant when Po0.05. Results The maximum depth of local anaesthetic was significantly less in the hyaluronidase group than the control group at 3 and 5 min (0.79 vs 1.65 mm, P-value 0.01 and 0.43 vs 1.52 mm, P-value 0.002 respectively). There were no statistically significant differences in the akinesia, pain and surgical satisfaction scores between the two groups. Conclusions The addition of hyaluronidase significantly augments the dispersal of local anaesthetic fluid, as measured by B-scan ultrasonography.

Bacterial vaginosis is the commonest cause of abnormal vaginal discharge in women of reproductive age and require laboratory test for diagnosis . A total 200 women aged 15-45 years with history of abnormal vaginal discharge were included as study population. Fifty women without such history of discharge were taken as healthy control. Three vaginal swab samples were taken from each case and control. These swab samples were subjected to test by conventional methods such as Amsel clinical criteria, Gram stain Nugent method, culture and by newly developed BV Blue test. The results of the BVBlue test were compared with these methods to find out the efficacy of BVBlue test. Rate of detection of bacterial vaginosis (BV) cases was 21.5% by Amsel clinical criteria, 21.0% by Gram stain Nugent method, 21.0% by culture and 22% by BVBlue test among the study population. When comparing with the conventional test and culture, BVBlue test was 100% sensitive and 98% specific. It is rapid, technically simple and is suitable for screening large number of patient in short time where laboratory facilities are not developed. Key words: Bacterial Vaginosis, BVBlue test, Nugent method, Abnormal vaginal discharge. DOI: http://dx.doi.org/10.3329/bjmm.v4i1.8465 BJMM 2011; 4(1): 24-27

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Degenerative and traumatic articular cartilage defects are common, difficult to treat, and progressive lesions that cause significant morbidity in the general population. There have been multiple approaches to treat such lesions, including arthroscopic debridement, microfracture, multiple drilling, osteochondral transplantation and autologous chondrocyte implantation (ACI) that are currently being used in clinical practice. Autologous bone-marrow mesenchymal cell induced chondrogenesis (MCIC) is a single-staged arthroscopic procedure. This method combines a modified microfracture technique with the application of a bone marrow aspirate concentrate (BMAC), hyaluronic acid and fibrin gel to treat articular cartilage defects. We reviewed the current literatures and surgical techniques for mesenchymal cell induced chondrogenesis.

Deficiency of the ninth component of human complement (C9) is the most common complement deficiency in Japan, with an incidence of approximately one homozygote in 1000, but is very rare in other countries. Genetic analyses of Japanese C9 deficiency have shown that a C-to-T transition leading to TGA stop codon for Arg95 in exon 4 of the C9 gene (Arg95Stop) is common in Japanese C9 deficiency. To determine the prevalence of heterozygous carriers of the Arg95Stop mutation in a Japanese population, we collected DNA samples from 300 individuals in two of the four main islands of Japan. Heterozygote detection was performed with an allele-specific polymerase chain reaction (PCR) system designed to detect exclusively only one of the normal and mutant alleles, followed by confirmation with PCR/single-strand conformation polymorphism (SSCP) analysis and direct sequencing. Twenty individuals were heterozygous for the Arg95Stop mutation. None was homozygous. The prevalence of carriers of the Arg95Stop mutation was 6.7% (20/300). An estimated frequency (0.12%) of complete C9 deficiency due to homozygous Arg95Stop mutation was consistent with frequencies determined by serological studies.