[Physiopathology of sudden death--pulmonary embolism].
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Biomedical subjects
Publications and source records attributed to H Fujioka.
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Rhoptries were isolated from merozoites of P. yoelii (17 XL), P. chabaudi adami and P. berghei (K-173), using sucrose gradient density centrifugation. Mouse antisera was prepared against the organelles and characterized. Antibodies specific for a known P. yoelii rhoptry protein were used to identify gradient fractions containing rhoptries and electron microscopy was used to confirm rhoptry enrichment and organelle morphology. Western blotting analysis of the gradients with organelle-specific antisera from each species, revealed several major cross-reactive interspecies protein bands of approximately 235, 210, 180, 160/170, 140, and 96-110 kDa, predominantly in densities of 1.12 and 1.15 g/ml. The parasite origin of the proteins was verified by immunoprecipitation, and reactive epitopes localized to the rhoptries by IEM. By Western blotting antisera specific for P. falciparum rhoptries reacted with protein bands of approximately 96-110 kDa in schizont extracts, and gradient fractions of density 1.12 and 1.15 g/ml from all three rodent malaria species, as well as with the rhoptries in P. yoelii, P. chabaudi, and P. berghei merozoites by IEM. We conclude that the three rodent malaria species and P. falciparum share conserved interspecies epitopes.
Thirteen monoclonal antibodies, obtained after immunization of mice with Plasmodium yoelii schizonts, were selected using immunofluorescence assay: they all presented typical fluorescence patterns of rhoptries. This antigen localization was confirmed by immunoelectron microscopy. The molecular weights of the recognized antigens are 68, 80, 105, 130 and 140 kDa as determined by immunoprecipitation and immunoblot under reducing and nonreducing conditions. These values are very similar to these of the low and high molecular weight complex components of Plasmodium falciparum. Furthermore, these antigens are soluble like P. falciparum rhoptry proteins. Interestingly, our monoclonal antibodies also reacted with two other Plasmodium species (Plasmodium berghei NKK173 strain and P. yoelii nigeriensis 798 VK strain), giving sometimes more complex labeling with apical, membranous, nuclear, or/and cytoplasmic localizations. Finally, none of the monoclonal antibodies stained the rhoptries of P. falciparum FCCE-1/Niger strain.
A systematic technique for the resection of hepatocellular carcinoma (HCC) prevents the dissemination of cancer cells through the portal vein of the remnant liver. We successfully performed a systematic laparoscopic left lateral segmentectomy in a 62-year-old man with HCC. The tumor was located in the left lateral segment of the liver, and measured approximately 4 cm in diameter. Since no other tumors were detected in the liver or in any distant organs, the patient was considered to be a candidate for surgery. A laparoscopic hepatic resection was selected as the procedure of choice. Prior to dissection of the liver parenchyma, the arteries and branches of the portal vein feeding the left lateral segment were divided and dissected, together with the branches of the biliary tree in the umbilical portion of the left pedicle of Glisson's capsule. The liver parenchyma was then dissected and the left hepatic vein divided and dissected, and transection of the left lateral segment was completed. The patient's postoperative course was uneventful and he was discharged on postoperative days 14. No evidence of recurrence has been noted in the 22 months after surgery (the time of this report). This less invasive surgery, taking into consideration the pathogenesis of HCC, may be a useful new approach in selected patients with this tumor.
PURPOSE: Approximately 15 to 20% of infertile men have azoospermia. In the Y chromosome a deletion, termed the azoospermic factor, has been found in some cases of idiopathic azoospermia. We investigate the relationship of factors in autosomal chromosomes (HLA class I antigens) to spermatogenesis failure in idiopathic azoospermia. MATERIALS AND METHODS: We evaluated 65 infertile Japanese men with idiopathic azoospermia. The frequency of the HLA allele reported in 1,216 healthy Japanese men was used as a control. HLA class I typing was performed by the National Institutes of Health standard serological method or polymerase chain reaction-sequence specific primer analysis. Allele frequencies were calculated. We determined statistical significance in the frequency of each allele in patients and controls using the chi-square test. The relationship of HLA antigens to idiopathic azoospermia was expressed as relative risk. RESULTS: In Japanese men with idiopathic azoospermia the frequency of HLA-A33, B13 and B44 was significantly increased compared with controls. The relative risk of HLA-B44 was 8.4, an extremely high value compared with that of other diseases and HLA antigens. CONCLUSIONS: We suggest that HLA class I antigens are important genetic markers that represent a risk factor for idiopathic azoospermia.
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Recent advances in technology have not substantially changed the high mortality rate associated with acute renal failure (ARF). To obtain a simple, valid prognostic index, we retrospectively evaluated the relative importance of demographic data, causes (acute insults) of renal failure, and comorbid clinical conditions for the outcome in 102 ARF patients who received renal replacement therapy with an overall mortality rate of 65% (66 of 102). There were no significant differences between survivors and nonsurvivors in age and gender. Mortality according to acute insults was similar to that of the whole population studied. Of the 10 clinical conditions at the time of the first renal replacement therapy, mechanical ventilation (p = 0.0002), cardiac failure (p = 0.0006), hepatic failure (p = 0.003), central nervous system dysfunction (p = 0.005), and oliguria (p = 0.04) were found to be significantly related to mortality by univariate analysis. Furthermore, multivariate analysis demonstrated that only mechanical ventilation, cardiac failure, and hepatic failure were significant risk factors. Survival was directly related to the number of significant variables in univariate analysis: zero, 89% (8 of 9); one, 62% (21 of 34); two, 19% (5 of 27); three, 10% (2 of 20); four, 0% (0 of 8); five, 0% (0 of 4). This simple and early prognostic index, derived from the assessment of clinical conditions which were easily determined at the patient's bedside, could be useful for outcome prediction in ARF patients requiring renal replacement therapy.
Despite its obscure and short effect, plasma exchange (PE) remains a mainstay in the treatment of liver disease. However, the question still remains as to whether or not PE suppresses the regeneration of the liver because PE deprives patients of hepatotrophic factors. The effect of PE, which could be a total blood exchange (TBE) in a syngeneic setting, on liver regeneration following a 68% partial hepatectomy (PH) was investigated in rats. In Group 1, 20 ml of blood from normal rats was infused while native blood was removed at 6 and 12 h after PH. In Group 2, 20 ml of blood obtained from PH rats at the same time points was infused. The regeneration rate, labeling index of proliferating cell nuclear antigen (PCNA), and plasma hepatocyte growth factor (HGF) level were determined, and standard liver function tests performed at 24, 48, and 72 h. Although all liver function tests improved in Group 1 at 24 and 48 h, the regeneration rate was significantly impaired. Similarly, the PCNA labeling index was significantly lower in Group 1 than that in Group 2. The plasma HGF level was significantly reduced in Group 1 (6 h blood out versus blood in: 1.1+/-0.5 vs. 0.1+/-0.1 ng/ml, p < 0.05). TBE with normal blood following PH suppressed the early stage of liver regeneration, in part, because of the reduction of HGF even though the blood was purified.
We have developed a new method to replace the conventional method of quantitatively measuring cerebral blood flow (CBF), in which octanol-extracted radioactivity counts are measured in continuous arterial blood samples. With the new method, the whole-brain time-activity curves early after the intravenous injection of N-isopropyl-p-[123I]iodoamphetamine were first obtained by the least-squares curve-fitting method. The equation thus obtained was differentiated and the radioactivity counts were corrected by the 5-min octanol-extracted radioactivity counts of the arterial blood sample obtained at a single time point. In the present study, the value obtained by integrating the equation from 0 to 5 min, which was obtained by curve-fitting, was compared with the octanol-extracted radioactivity counts obtained by 5-min continuous arterial blood sampling in 160 patients with cerebrovascular disorders. The results showed good agreement between the values obtained by the two procedures (y = 1.049x - 1522.4; r = 0.987). Using the CBF obtained by the 5-min continuous arterial blood sampling as the standard, the errors using the present integral values of the input function were 7.1 +/- 4.9%. Measurements of the integral of the input function by the present one-point arterial blood sampling method has the potential for use in the routine measurement of CBF, because it is less invasive and more convenient than the conventional method, and it is unaffected by cardiopulmonary disease or smoking.
In addition to its potent efficacy in animal models against Candida sp., Aspergillus fumigatus, and Histoplasma capsulatum, the clinical candidate pneumocandin MK-991 (formerly L-743,872) was also extremely potent against Pneumocystis carinii in models of immune-compromised animals. MK-991 was approximately 14 times more potent than the original natural product lead, pneumocandin B0. The 90% effective dose (ED90) of MK-991 for cyst clearance in the rat model for pneumocystis was 0.011 mg/kg of body weight when delivered parenterally for 4 days twice a day (b.i.d.). In a mouse model, under the same experimental parameters, the ED90 was 0.02 mg/kg. MK-991 was also effective orally, with an ED90 for cyst clearance of 2.2 mg/kg against acute infection in rats (b.i.d. for 4 days). Complete prevention of P. carinii development was achieved in immunocompromised mice at a daily oral dose of 2.25 mg/kg. As reported previously for other pneumocandins and echinocandins, MK-991 selectively prevented the development of P. carinii cysts. When used as a prophylactic agent, neither stage of the organism appeared in the lungs of animals. In response to an acute infection, cysts were eliminated rapidly, while trophozoite forms persisted. Despite good efficacy as an oral agent in murine models, the low oral absorption of this class may limit the use of MK-991 to parenteral therapy.
The mei4+ gene of the fission yeast Schizosaccharomyces pombe was cloned by functional complementation. The mei4 disruptant failed to complete meiosis-I but could proliferate normally. mei4+ was transcribed only in meiosis-proficient diploid cells after premeiotic DNA replication. The mei4+ open reading frame encodes a 57-kDa serine-rich protein comprised of 517 amino acids with a forkhead/HNF3 DNA-binding domain in the amino-terminal region. Transcription of spo6+, a gene required for sporulation, was dependent on the mei4+ function. Two copies of the GTAAAYA consensus sequence, proposed as the binding site for human forkhead proteins, were found in the promoter region of spo6+. A gel mobility shift assay demonstrated the sequence-dependent binding of the GST-Mei4 forkhead domain fusion protein to DNA fragments with one of the consensus elements. Deletion of this consensus element from the spo6 promoter abolished the transcription of spo6+ and resulted in a sporulation deficiency. One-hybrid assay of Mei4 which was fused to the Gal4 DNA-binding domain localized the transcriptional activation domain in the C-terminal 140 amino acids of Mei4. These results indicate that Mei4 functions as a meiosis-specific transcription factor of S. pombe.
In isolated canine cerebral and cutaneous arteries and duodenum, effects of neosurugatoxin (NSTX) on the response to nicotine were compared. Nicotine-induced relaxations are mediated by nitric oxide (NO) from vasodilator nerves in cerebral arteries and by calcitonin gene-related peptide (CGRP) in cutaneous arteries treated with NO synthase inhibitors. Duodenal relaxation to nicotine is mediated by NO from inhibitory nerves. Cerebral arterial strips without endothelium responded to nicotine with relaxations that were inhibited by NSTX (3 x 10(-10) to 3 x 10(-9) M) concentration-dependently. Relaxations to nicotine of duodenal strips were attenuated by NSTX and abolished by tetrodotoxin, whereas those induced by K+ and electrical stimulation were not influenced. In cutaneous arteries treated with N(G)-nitro-L-arginine, nicotine-induced relaxations were attenuated by NSTX as low as 10(-10) M, but unaffected by tetrodotoxin. The inhibitory potency of NSTX was in the order of cutaneous artery > duodenum > cerebral artery. It is concluded that NSTX selectively antagonizes actions on nicotinic receptors in nitroxidergic nerves of cerebral arteries, CGRP-mediated sensory nerves of cutaneous artery and ganglionic cells of the duodenum, but the affinity of this toxin to nicotinic receptors in sensory nerves is the highest.
The effects of surgical repair versus non-repair on cell morphology and type X collagen expression were investigated using a rat model of Achilles tendon avulsion. The animals were divided into four groups. In Group 1, tendon was reattached to the original attachment site by suturing through a drill hole in the calcaneus; in Group II, tendon was not reattached and a drill hole was not made; in Group III, tendon was not reattached but a drill hole was made; and the animals in Group IV were sham operated. In Group I (tendon reattached), at 2 weeks postoperatively, many hypertrophic chondrocytes appeared at the reattachment site adjacent to bone and type X collagen was detected immunologically both in the cells and in the extracellular matrix. After 4 weeks, the cells at the original site of attachment were arranged in rows along the newly formed tendon fibers and were stained with type X collagen antibody. By contrast, when tendon was not reattached (Groups II and III), a gap between the original attachment site and the tendon stump was observed through the entire postoperative period. At 8 weeks, the original attachment site was covered by fibrocartilaginous tissue and tendon became attached to the calcaneal fibrocartilage area, which is proximal to the original attachment site. Type X collagen was detected in the cells which were adjacent to bone. In Group IV (sham operation), there were no changes in histology or type X collagen distribution, either at the attachment site or in tendon and bone, compared with the non-operated control rats. These results suggest that surgical reattachment of tendon to the original site is important to help reorganize cells during the repair process. Type X collagen was identified immunohistochemically in the cells adjacent to bone in all the groups, suggesting that it may play a role in maintaining distinct areas of calcified and non-calcified fibrocartilage.
PURPOSE: We determined a rational strategy for treatment of patients with retrograde ejaculation in the era of modern assisted reproduction technology. MATERIALS AND METHODS: In 7 consecutive patients medical treatment or retrieval of spermatozoa from the bladder was performed at a male infertility clinic. RESULTS: Antegrade ejaculation was restored in 3 patients, and spermatozoa were retrieved from the bladder and used for assisted reproduction in 3. Spermatozoa with good oolemma penetrating ability were collected by seminal vesicle massage. CONCLUSIONS: Modern assisted reproduction technology is a powerful treatment option for retrograde ejaculation when combined with a technique to retrieve spermatozoa of good quality from the bladder.
Cerebral blood flow (CBF) has been measured using a microsphere model with octanol-extracted radioactivity counts (integral value of input function). We developed a new method estimating the integral value of input function. First, we fitted the whole brain time-activity curves early after intravenous injection of N-isopropyl-p-[123I]iodoamphetamine (123I-IMP) by the least-squares method. Second, we differentiated this equation. Third, we calibrated it using octanol-extracted radioactivity counts of the arterial blood sampled at 5 min. Finally, we integrated it. A significant correlation was found between the integral values obtained using a new method and those obtained using the continuous arterial blood sampling data (y = 1.048x-1206, r = 0.984). The errors between the CBF values obtained using a new method and those obtained using the 5-min continuous arterial blood sampling was 6.88 +/- 4.78%. Measurement of integral values of the input function using a new method with one-point arterial blood sampling is less invasive and convenient, and is not influenced by cardiopulmonary disease or smoking. Therefore, it would be useful for the routine measurement of CBF.
We developed a new microsphere method using 123I-IMP in which arterial blood is collected at one time point early after 123I-IMP injection instead of conventional continuous arterial blood sampling, and the input count is estimated using a whole brain time-activity curve until that time point. The differential curve dCb(t)/dt of the brain time-activity curve Cb(t) early after 123I-IMP injection (when the microsphere model is established) approximates the arterial time-activity curve Ca(t) as an input function. The input count as the integral value of Ca(t) at 0-5 min was estimated using the 50 min whole brain activity Cb(5 min) and the differential value dCb(5 min/dt and the 5-min octanol-extracted count of arterial samples obtained at one time point Ca(5 min). The input counts calculated by this method were very closely correlated with measurement values obtained by continuous arterial sampling. This method is more convenient and less invasive than the continuous arterial sampling method.