Cell cycle control and CDC28/Cdc2 homologue and related gene cloning of Cryptococcus neoformans]

In Cryptococcus neoformans the DNA content of cells having tiny buds varied rather widely, depending on growth phases and strains used. Typically, buds of C. neoformans emerged soon after initiation of DNA synthesis in the early exponential phase. However, bud emergence was delayed to G2 during transition to the stationary phase, and in the early stationary phase budding scarcely occurred, although roughly half of the cells completed DNA synthesis. The timing of budding in C. neoformans was shifted to later cell cycle points with progression of the growth phase of the culture. Similarly, a deficit in oxygen was demonstrated to delay the timing of budding, prolong the G2 phase and cause accumulation of cells after DNA synthesis, but before commitment to budding. The C. neoformans homologue of the main cell cycle control gene CDC28/Cdc2 was isolated using degenerate RT-PCR. The full-length coding region was then amplified using primers to target the regions around the start and stop codons. The gene was called CnCdk1 and was found to have high homologies to S. cerevisiae CDC28 and S. pombe cdc2. To determine its function, its ability to rescue S. cerevisiae cdc28-temperature sensitive mutants was tested. S. cerevisiae cdc28-4 and cdc28-1N strains transformed with the pYES2-CnCdk1 construct exhibited growth at the restrictive temperature. Results of the sequence analysis and the ability of CnCdk1 to complement the S. cerevisiae cdc28-ts mutations support its assumed role as the CDC28/cdc2 homologue in C. neoformans.     

Marchiafava-Bignami disease treated by mianserin hydrochloride in short-term evaluated by neuropsychological analysis

Since 1903, Marchiafava‐Bignami disease has been recognized as a rare syndrome with focal demyelination and necrosis in the corpus callosum, which is usually found in chronic alcoholics. It extends into the neighboring white matter and occasionally as far as the subcortical regions. We report a Japanese patient with Marchiafava‐Bignami disease associated with alcohol abuse, who had traveled around Western Europe, North America and China for more than 30 years. As he suffered extreme delirium in the early stages we administered a low dose (10 mg) of mianserin hydrochloride. He was very irritable and uncooperative on admission, after 20 days his delirium had disappeared and his temper had become very calm and mild. After 40 days, his intelligence level increased substantially as measured by various neuropsychological tests.     

Ultrasound registration of the bone surface for surgical navigation.

OBJECTIVE: To allow non-invasive registration of the bone surface for computer-assisted surgery (CAS), this investigation reports the development and evaluation of intraoperative registration using 2D ultrasound (US) images. This approach employs automatic segmentation of the bone surface reflection from US images tagged with the 3D position to enable the application of CAS to minimally invasive procedures. METHODS: The US-based registration method was evaluated in comparison to point-based registration, which is the predominant method in current clinical use. The absolute accuracy of the US-based registration was determined using a phantom pelvis, with fiducial registration providing the ground truth. The relative accuracy was determined by an intraoperative study comparing the US registration to the point-based registration obtained as part of the HipNav experimental protocol. RESULTS: The phantom pelvis study demonstrated equivalent accuracy between point- and US-based registration under in vitro conditions. In the intraoperative study, the US-based registration was sufficiently consistent with the point-based registration to warrant larger-scale clinical trials of this non-invasive registration method. CONCLUSION: Ultrasound-based registration eliminates the need for physical contact with the bone surface as in point-based registration. As a result, non-invasive registration could fully unlock the potential of computer-assisted surgery, enabling development of the next generation of minimally invasive surgical procedures.     

Metabolism of 99mTc-ethyl cysteinate dimer (99mTc-ECD) in blood--mainly in vitro]

Metabolism of 99mTc-ethyl cysteinate dimer (99mTc-ECD) in blood was studied mainly in vitro. When 99mTc-ECD was mixed with blood taken from 12 subjects, the octanol extraction ratio of ECD (y) decreased rapidly and the octanol extraction ratio-time profile well fitted a monoexponential curve (y = Ae-kt/1000, A, k: constant, t: time after mixing). The k value and hematocrit (Ht) were significantly correlated (k = 0.376Ht-3.27, r = 0.897, p less than 0.001), therefore, it was suggested that the majority of the enzyme which dissolves ECD exists in red blood cells. When ECD was mixed with blood, there were more hydrophilic products of ECD in plasma than those generated by the enzyme in plasma. In vivo input function of 99mTc-ECD was calculated by arterial blood sampling and octanol extraction. The duration of effective input was relatively short, which was attributed to rapid decrease of octanol extraction ratio in vivo.     

Two cases of persistent hypouricemia associated with diabetes mellitus.

Two patients with diabetes mellitus had persistent hypouricemia due to increased urate clearance; the degree of the apparent renal hypouricemia with uricosuria was quite mild. At the onset of diabetes, their serum urate levels were normal. Even after good diabetes control in both cases, hypouricemia continued. Based on the pharmacological evaluation in both patients, pyrazinamide administration could partially decrease urate clearance, however, suppression by pyrazinamide was less than in normal subjects, and probenecid increased urate clearance. These results suggest that the present cases had a renal abnormality affecting tubular presecretory reabsorption of urate, which might be due to diabetes mellitus.     

Posterior instability near extension is related to clinical disability in isolated posterior cruciate ligament deficient patients

The purpose of this study is to evaluate the relationship between the magnitude of knee laxity and posterior instability at different knee flexion angles and clinical disability in isolated posterior cruciate ligament (PCL) deficient patients. Knee laxity at 20° and 70° of knee flexion were evaluated using KT-2000 arthrometer, and the posterior instability at 20°, 45° and 90° of flexion were evaluated using stress radiography. We assessed the differences in the knee laxity and the tibial translation between isolated PCL deficient knees and normal knees, and between the patients with giving-way during activities of daily living (ADL) and without giving-way. There were statistical differences in the knee laxity and the tibial translation at all knee flexion angles between the PCL deficient knees and normal knees. The magnitude of the knee laxity at 20° of flexion measured with KT-2000 arthrometer was significantly larger in the patients with giving-way than those in the patients without giving-way although there was no significant difference in the tibial translation at 70° between the two groups. The tibial translation in both medial and lateral compartments at 20° and 45° measured with stress radiography were significantly larger in the patients with giving-way than those in the patients without giving-way although there was not significant difference at 90° between the two groups. These results suggested that the magnitude of the knee laxity and the posterior tibial translation at shallow knee flexion angles would be related to giving-way during ADL in isolated PCL deficient patients.