Distinction of Paramecium strains by a combination method of RAPD analysis and multiplex PCR.

Paramecium is employed as a valuable model organism in various research fields since a large number of strains with different characteristics of size, morphology, degree of aging, and type of conjugation can be obtained. It is necessary to determine a method for the classification and simple identification of strains to increase their utility as a research tool. This study attempted to establish a polymerase chain reaction (PCR)-based method to differentiate strains of the same species. Genomic DNA was purified from several strains of P. caudatum, P. tetraurelia, and P. bursaria used for comparison by the random amplified polymorphic DNA (RAPD)-PCR method. In P. tetraurelia and P. bursaria, it was sufficiently possible to distinguish specific strains depending on the pattern of random primers and amplification characteristics. For the classification of P. caudatum, based on the sequence data obtained by RAPD-PCR analysis, 5 specific primer sets were designed and a multiplex PCR method was developed. The comparative analysis of 2 standard strains, 12 recommended strains, and 12 other strains of P. caudatum provided by the National BioResource Project was conducted, and specific strains were identified. This multiplex PCR method would be an effective tool for the simple identification of environmental isolates or the management of Paramecium strains.

Comparative Analysis Between Paramecium Strains with Different Syngens Using the RAPD Method.

Paramecium spp. are a genus of free-living protists that live mainly in freshwater environments. They are ciliates with high motility and phagocytosis and have been used to analyze cell motility and as a host model for pathogens. Besides such biological characteristics, apart from the usual morphological and genetic classification of species, the existence of taxonomies (such as syngens) and mating types related to Paramecium's unique reproduction is known. In this study, we attempted to develop a simple method to identify Paramecium strains, which are difficult to distinguish morphologically, using random amplified polymorphic DNA (RAPD) analysis. Consequently, we can observe strain-specific band patterns. We also confirm that the presence of endosymbiotic Chlorella cells affects the band pattern of P. bursaria. Furthermore, the results of the RAPD analysis using several P. caudatum strains with different syngens show that it is possible to detect a band specific to a certain syngen. By improving the reaction conditions and random primers, based on the results of this study, RAPD analysis can be applied to the identification of Paramecium strains and their syngen confirmation tests.

[Usefulness of ultrasound sonography in performing posterior sacroiliac ligament injection in elderly patients].

BACKGROUND: The sacroiliac joint (SIJ) is recognized as a potential source for low back pain especially in elderly people. Previous studies indicated that in patients with SIJ pain, posterior sacroiliac ligament injections are more effective than intra articular injections. The aim of this study was to assess usefulness of ultrasound sonography (US) in performing posterior sacroiliac ligament injection. METHODS: We examined SIJ and ligaments, and injected local anesthetics using ultrasonography. US guided needle insertions were performed. We also examined the cadevar of a 74-year-old woman to examine SIJ anatomy. RESULTS: US was performed in 61 male and 97 female outpatients (N = 158) with a mean age of 72.8 ± 10.9. US examination of the SIJ is difficult to perform because of its complex anatomy, and wide range of normal variations, and relative inaccessibility by surface scanning. The long posterior sacroiliac ligament (LPSL) is visualized as a hyper echoic laminated linear structure. We were able to confirm needle placement and area affected by local anesthetics. CONCLUSIONS: Our study suggests that US can be useful in performing posterior sacroiliac ligament injections. Sufficient anatomical knowledge and extensive US experience are essential in performing these injections.

[Effect of the sacroiliac ligament block on intractable low back pain in elderly patients].

BACKGROUND: Lumbar spine disease in the elderly people is complicated by a variety of pathophysiology in the spine and the cause of the pain is unclear. Diagnosis of pain may be difficult in patients with pain in the thigh and groin area. Sacroiliac joint is supporting the trunk and movable joint. We examined the effect of the sacroiliac joint block for intractable low back pain. METHODS: Retrospectively we examined the duration of disease in patients with hip and leg pain visiting the hospital for eight months, and we questioned the site of pain awareness. Newton test, Gaenslen test, Patrick test and Fadire test were carried out for sacroiliac joint pain in patients with at least one positive finding. When performing sacroiliac ligaments block local anesthetics was injected to check the position of the dorsal sacroiliac ligaments under ultrasonic echo whenever possible. The block is performed with the patients prone at a point one finger from the posterior superior iliac spine level at an angle of 30-45 degrees downward toward the outside. Injecting the drugs penetrating the ligament continued to give a feel slightly outward to avoid the iliac Cattelan 23 G needle. We confirmed pain assessment NRS at 11 (0-10), and the improvement of pain was assessed with the change of the NRS on the next return. RESULTS: NRS showed a significant decrease at all points in time before block, their pain decreased gradually. The patients showed NRS improvement of more than 50% of the first block in 15 of 24 patients. CONCLUSIONS: Block at the posterior sacroiliac ligament region significantly reduced pain for chronic intractable low back pain. The block was shown to be effective as a treatment and for diagnosis.

[Anesthetic management using auto-transfusion and hypothermia for massive bleeding].

A 51-year-old woman was scheduled for emergency enterectomy and vascular repair under general anesthesia for active bleeding from internal iliac artery caused by repeated radiotherapy for cervical cancer and subsequent hypovolemic shock. For the first two hours of operation, the blood loss exceeded 6,000 ml and the hemoglobin level decreased to a low of 3.8 g x dl(-1) despite administration of 38 units of packed red cells. Intraoperative blood salvage was used in order to minimize further loss of hemoglobin. Mild hypothermia technique was also introduced to prevent brain ischemia. Total bleeding volume was approximately 10,000 ml, and total transfused volume was 8,740 ml. No neurological deficit and no systemic infection were found during the postoperative course. Although clinical risks of cell salvage in patients undergoing surgery for malignant tumor remain controversial, we conclude intraoperative blood salvage using Cell Saver could be utilized as a life-saving means and mild hypothermia might have been efficacious for protecting the brain from ischemia in our case.