Identification of 66 kDa phosphoprotein associated with motility initiation of hamster spermatozoa

Background and Aims:  Sperm motility is regulated by protein phosphorylation. The 66 kDa protein obtained from hamster sperm flagella was phosphorylated at serine residues associated with the motility initiation. In order to understand the regulatory mechanism of sperm motility, the 66 kDa protein was identified in the present study.
Methods:  The 66 kDa protein was purified by 2-D gel electrophoresis and identified by matrix-assisted laser desorption ionization mass spectrometry, liquid chromatography-tandem mass spectrometry and peptide sequencer.
Results:  The 66 kDa protein was tubulin β chain.
Conclusion:  The 66 kDa protein is one of the tubulin β chain isoforms and phosphorylated in relation to the motility initiation. (Reprod Med Biol 2004; 3 : 133–139)     

Detection of glyceraldehyde 3-phosphate dehydrogenase messenger RNA using a peptide nucleic acid probe in paraffin-embedded archival specimens

Although the human genome project has been completed, the functions of many genes remain undetermined. In situ hybridization (ISH) is a key method for identifying cells in which a given messenger RNA is transcribed. Paraffin-embedded specimens remain precious materials for research, but preservation of high-quality RNA in these specimens is not expected unless ample caution was taken during fixation. Peptide nucleic acid (PNA) is a recently developed hybrid molecule with genetic information that has high stability and high affinity to the complementary DNA or RNA. We applied a PNA probe to mRNA ISH of liver specimens obtained by autopsy and embedded in paraffin 28-48 years ago. An 18-mer PNA probe for glyceraldehyde 3-phosphate dehydrogenase was used. Staining was then analyzed in association with morphology by hematoxylin and eosin staining, and with the time between death of the patient and tissue fixation. Notably, specimens fixed with formalin and embedded in paraffin 48 years ago yielded excellent results if the time before fixation was short enough (<8 h). There was a significant inverse correlation between the intensity of ISH staining and the time before fixation. Oligonucleotide PNA probe, albeit at high cost, would increase the value of paraffin-embedded specimens in storage for use in human medical research.     

Zinc deficiency in patients with idiopathic taste impairment with regard to angiotensin converting enzyme activity

OBJECTIVE: The present study aimed at measuring the ratio of apo/holo activities of angiotensin converting enzyme (ACE) ratio in the serum of patients with taste impairment to evaluate their status of zinc nutrition. METHODS: Nineteen patients complaining of taste impairment were divided into two groups: zinc-deficiency taste impairment (n=6) and idiopathic taste impairment (n=13) and compared to 30 volunteers. Zinc concentrations in the serum were measured by atomic absorption spectrometry (normal values: 64-111 microg/dl). Patients with zincemia values of <63 microg/dl with no history of other disorder or medication known to cause dysgeusia were diagnosed as zinc deficient, while those with the same condition and values >64 microg/dl were considered to belong to the idiopathic group. The activities of ACE in the serum were measured spectrophotometrically as the activity of the holo-ACE, and after addition of 80 microM of zinc to the serum in vitro, the increase of ACE activity over initial value in the serum was determined as that of the apo-ACE. Finally, the apo/holo-ACE activities ratio was used as an index of zinc nutritional status. RESULTS: The mean concentrations of zinc in the serum were 77.4+/-8.4 microg/dl in volunteers, 77.6+/-8.4 microg/dl in the idiopathic patients and significantly decreased at 55.7+/-5.8 microg/dl in zinc-deficiency patients. ACE activities in the serum were 14.7+/-7.6, 14.5+/-4.0 and 14.1+/-3.3 IU/l in volunteers, the idiopathic group and zinc-deficiency taste impairment group, respectively. The mean ACE ratios were 1.10+/-0.6% in volunteers and significantly increased at 9.8 +/- 4.0% in the idiopathic group and at 13.7+/-6.6% in zinc-deficiency taste impairment group. CONCLUSION: The results of the present study indicated that zinc deficiency is a predominant factor underlying hypogeusia even when zinc concentrations are within normal ranges in the serum and show that clinically, ACE ratio may be a more sensitive indicator of the zinc nutritional status than measuring zinc concentration in the serum.     

Sotalol-induced coronary spasm in a patient with dilated cardiomyopathy associated with sustained ventricular tachycardia

A 54-year-old man with severe left ventricular dysfunction due to dilated cardiomyopathy was referred to our hospital for symptomatic incessant sustained ventricular tachycardia (VT). After the administration of nifekalant hydrochloride, sustained VT was terminated. An alternate class III agent, sotalol, was also effective for the prevention of VT. However, one month after switching over nifekalant to sotalol, a short duration of ST elevation was documented in ECG monitoring at almost the same time for three consecutive days. ST elevation with chest discomfort disappeared since he began taking long-acting diltiazem. Coronary vasospasm may be induced by the non-selective beta-blocking properties of sotalol.     

Successful management of intractable epilepsy with lidocaine tapes and continuous subcutaneous lidocaine infusion

PURPOSE: We report the successful management of a 10-year-old girl with intractable frontal lobe epilepsy by using lidocaine tapes and continuous subcutaneous lidocaine infusion. METHODS: This patient's seizures were refractory to conventional antiepileptic drugs (AEDs) and mexiletine, but they responded well to the intravenous infusion of lidocaine. The intravenous infusion of lidocaine was replaced by lidocaine tape therapy, and subsequently by continuous subcutaneous lidocaine infusion therapy. The lidocaine tape (Penles, Nihon Lederle, Tokyo Japan) used was a stamp-sized (30.5 x 50.0 mm) tape containing 18 mg of lidocaine. We used 25 lidocaine tapes every 12 h (50 tapes/day). Lidocaine hydrochloride (10%) was administered continuously at a dose of 1.5 mg/kg/h (0.3 ml/hour) through a 27-G needle that was inserted in the subcutaneous tissue. RESULTS: Lidocaine tape therapy showed good efficacy for 1 year. After that, six lidocaine tapes were added 6 h after the exchange of 25 lidocaine tapes [62 tapes/day (25,6,25,6)], because the seizures became frequent when the lidocaine tapes were being exchanged. The seizures were then well controlled, but dermatitis due to the lidocaine tapes grew serious, and lidocaine tape therapy had to be stopped. Continuous subcutaneous infusion of lidocaine applied in place of lidocaine tapes provided long-term seizure control without remarkable side effects. CONCLUSIONS: Lidocaine tape therapy and continuous subcutaneous lidocaine infusion therapy were considered to be useful for controlling this patient's seizures. This is the first report to describe the efficacy of continuous subcutaneous lidocaine infusion therapy for epilepsy.     

Preparation of antibacterial silver-doped silica glass microspheres

Various types of inorganic substances doped with silver ions have been developed as antibacterial materials, and some have already been commercialized. Colorless and chemically durable materials that slowly release silver ions are, however, still need to be developed. The present authors have previously shown that when a silica glass doped with silver and aluminium ions is prepared using the sol-gel method, the resultant product is colorless, chemically durable, and slowly releases silver ions into water over a long period. The doped silica glass takes a form of microspheres <1 microm in diameter, it is easily mixed with organic polymers, and the mixture can be formed into a thin film or fine fibers, etc. We report on the preparation of silver doped silica glass microspheres having a diameter =1 microm, using the sol-gel method. Initially, tetraethoxysilane was partially prehydrolyzed by water in ethanol, and then aluminium triisopropoxide was added to the solution to form Si-O-Al bonds. Finally, an ammonia solution containing silver nitrate was added to form silica microspheres doped with silver ion together with aluminium ions. The results show monodispersed microspheres 0.4-0.6 microm in diameter were obtained with nominal compositions of Si/Al/Ag = 1/0.01-0.03/0.003-0.03, with a molar ratio of Al/Ag = 1-3.3. The microspheres were colorless, showed a high chemical durability, and slowly released silver ions into water at 37 degrees C. Microspheres with the composition Si/Al/Ag = 1/0.01/0.01 showed excellent antibacterial activity against Escherichia coli. The minimum inhibitory concentration (MIC) of the microspheres was 400, which is less than the MIC value (800) of commercial antibacterial materials.     

Changes in left anterior descending coronary artery flow profiles after coronary artery bypass grafting examined by means of transthoracic Doppler echocardiography

OBJECTIVE: We sought to investigate the changes of velocity profiles in the left anterior descending coronary artery after coronary artery bypass grafting using transthoracic Doppler echocardiography. METHODS: Forty-five patients who received a bypass graft to the left anterior descending coronary artery were studied. Before coronary artery bypass grafting, Doppler velocity profiles of the distal left anterior descending coronary artery were recorded with transthoracic Doppler echocardiography. Peak systolic velocity, mean systolic velocity, peak diastolic velocity, mean diastolic velocity, total velocity time integral, systolic velocity time integral, and diastolic velocity time integral were measured. Three weeks after coronary artery bypass grafting, left anterior descending coronary artery antegrade flow in the distal portion of the anastomosis was obtained by using the same method. Coronary angiography was performed before and 3 weeks after coronary artery bypass grafting. RESULTS: The overall success rate of measuring the left anterior descending coronary artery flow was 60.0% preoperatively and 80.0% postoperatively. In 25 patients, in whom all parameters were obtained both before and after coronary artery bypass grafting, the following increased significantly after coronary artery bypass grafting: peak systolic velocity (14.86 +/- 7.50 vs 25.07 +/- 17.02 cm/s, P =.0045), mean systolic velocity (9.86 +/- 5.42 vs 18.03 +/- 12.94 cm/s, P =.0026), peak diastolic velocity (24.26 +/- 12.54 vs 48.28 +/- 31.66 cm/s, P =.0021), mean diastolic velocity (14.94 +/- 6.65 vs 30.36 +/- 20.71 cm/s, P =.0022), diastolic velocity time integral (7.22 +/- 2.88 vs 15.55 +/- 10.39 cm, P =.0009), total velocity time integral (10.50 +/- 4.48 vs 19.27 +/- 12.63 cm, P =.0034), and diastolic-to-systolic velocity time integral ratio (3.09 +/- 1.53 vs 4.97 +/- 2.75, P =.0044). Angiography showed graft patency and no significant change in left anterior descending coronary artery stenosis in all patients. CONCLUSIONS: Transthoracic Doppler echocardiography showed a significant increase in some parameters in left anterior descending coronary artery flow after coronary artery bypass grafting. Measurement of left anterior descending coronary artery flow by means of transthoracic Doppler echocardiography might be a noninvasive method to evaluate the effect of bypass grafting on the left anterior descending coronary artery.     

Homocysteine and hemostatic disorder as a risk factor for myocardial infarction at a young age

Introduction: Hyperhomocysteinemia is a coronary risk factor, but its pathophysiologic mechanism remains unclear. Materials and methods: The importance of hyperhomocysteinemia in the pathogenesis of early myocardial infarction, was determined in case-control study of 127 men with a first early myocardial infarction ≤45 years and 150 age-matched male controls. We measured plasma concentrations of homocysteine, fibrinogen, antithrombin, tissue factor, tissue factor pathway inhibitor, tissue plasminogen activator, plasminogen activator inhibitor-I, plasminogen, ₂-antiplasmin, lipoprotein(a), protein C, protein S, factor VII, and activated factor VII. Results: Homocysteine concentrations were higher in patients with early myocardial infarction than in controls (11.2±5.3 and 8.3±5.0 μmol/l, respectively, P<0.001). Hyperhomocysteinemia was associated with early myocardial infarction (odds RATIO=2.22, P<0.001) by multivariate logistic regression analysis. Tissue factor, antithrombin, plasminogen, tissue plasminogen activator, plasminogen activator inhibitor-I, lipoprotein(a), diabetes, and smoking also had associations. In a stepwise logistic regression analysis, hyperhomocysteinemia was the strongest predictor of early myocardial infarction (R²=0.19, P<0.001). Hyperhomocysteinemia also had positive correlations with tissue factor (ρ=0.26, P=0.009), tissue factor pathway inhibitor (ρ=0.23, P=0.020), and tissue plasminogen activator (ρ=0.25, P=0.011) in patients with early myocardial infarction, but not in controls. Conclusions: Hyperhomocysteinemia is an independent risk factor for early myocardial infarction, and is associated with a hypercoagulable state mediated by the extrinsic coagulation cascade.