Percentages of water, muscle, and bone decrease and lipid increases in early period after successful kidney transplantation: a body composition analysis.

BACKGROUND: Successful kidney transplantation (KT) can theoretically reconstitute body composition of a patient with chronic kidney disease (CKD). However, the practical changes have not been well documented. We evaluated changes in body composition among candidates before and 1 year after KT. METHODS: We enrolled 37 male and 18 female kidney recipients eligible for comparison of their body mass index (BMI), body composition, and lipid metabolism before and 1 year after KT. Twenty-one patients had been induced with a calcineurin inhibitor, mycophenolate mofetil, steroid, and basiliximab, and 34 others underwent steroid withdrawal on postoperative day 3. The body composition was analyzed using bioelectrical impedance. We also analyzed changes in BMI and lipid profiles. RESULTS: There was no significant change in BMI (21.4 ± 3.1 vs 21.7 ± 3.5 kg/m(2)). Regarding body composition, the water level decreased significantly (61.2 ± 4.9% vs 58.3 ± 5.3%; P < .05). In contrast, fat significantly increased (16.4 ± 6.7% vs 20.3 ± 7.1%; P < .05). More interestingly, successful KT significantly decreased the muscle and bone mass at 1 year after KT (37.3 ± 5.1% vs 34.8 ± 4.7%; 16.3 ± 2.1% vs 15.2 ± 2.1%; respectively; P < .05). Serum lipid profiles of total cholesterol, low-density lipoprotein cholesterol, and triglyceride worsened after KT. Comparing the 2 protocols, there was no difference in any item. CONCLUSIONS: Care must be taken even after successful KT to avoid dyslipidemia, which is a risk factor for cardiovascular disease. Well programmed dietary and/or exercise protocols to prevent muscle atrophy and fat gain should be considered even after successful KT.

Influence of calcium channel blockers in patients with gastrointestinal disease in Japanese community pharmacies.

WHAT IS KNOWN AND OBJECTIVE: Calcium channel blockers (CCBs), which have been widely used for the treatment of hypertension and angina pectoris, decrease lower oesophageal sphincter pressure and, as a result, can exacerbate gastrointestinal disease. In a previous study, increased risk of exacerbation of gastrointestinal disease among elderly patients following treatment with CCBs was identified. The prevalence of gastrointestinal diseases has increased in elderly patients, and it is possible that treatment with CCBs may have undesirably influenced this increase. The change in risk of gastrointestinal disease can be estimated by analysing changes in the prescription of antisecretory drugs as an outcome of exacerbation of gastrointestinal disease caused by CCBs. METHODS: It was hypothesized that patients who were prescribed CCBs would also change their use of antisecretory drugs. From September 2005 to August 2009, a dynamic retrospective cohort study was performed at five community pharmacies in Nagasaki, Japan, to assess alteration of antisecretory drug therapy following treatment with CCBs. Correlations with alterations of antisecretory drug therapy were determined by the Cox proportional hazards model. RESULTS AND DISCUSSION: The proposed study included 260 patients who were prescribed CCBs and 155 controls. During the study period, 53 patients were prescribed CCBs and 13 controls altered their antisecretory drug therapy; the hazard ratio was 2·22 (95% CI 1·25-4·26). WHAT IS NEW AND CONCLUSION: Calcium channel blocker treatment of patients with gastrointestinal disease was associated with alteration in frequency of prescription and an increase in dosage of antisecretory drugs. For clinical management of hypertension, alternative antihypertensive drugs may be considered for patients with gastrointestinal diseases. Further studies are required to determine the influence of CCB therapy on gastroesophageal diseases, suggested by the increase in use of antisecretory drugs.

Conteúdo polifenólico e atividade antioxidante dos frutos da palmeira Juçara (Euterpe edulis Martius) / Polyphenolic content and antioxidant capacity of fruits of Juçara (Euterpe edulis Martius) palm tree

O conteúdo polifenólico e a atividade antioxidante do extrato do fruto de Euterpe edulis Martius foram avaliados. Esta espécie é uma palmeira conhecida como Juçara, nativa da Mata Atlântica e utilizada para a extração de palmito. O processo de extração do palmito acarreta a morte da planta, uma vez que esta apresenta estipe único. A elevada demanda ocasionou a escassez deste recurso natural. Muitas espécies da Mata Atlântica podem ser utilizadas pelo manejo sustentável para a preservação e exploração econômica pelas comunidades locais. O fruto da palmeira Juçara pode ser uma das alternativas de manejo sustentável dos recursos naturais da Mata Atlântica. A capacidade antioxidante do fruto pode ser utilizada como justificativa para a aplicação como alimento nutricional. O conteúdo polifenólico do fruto foi determinado pelo método de Folin-Ciocalteau e os resultados obtidos foram: 10,31 ± 0,25%, 12,42 ± 0,89%, 12,75 ± 0,94%, para o extrato bruto, fração acetato de etila e fração remanescente, respectivamente. A atividade antioxidante foi determinada pelos métodos de redução do complexo fosfomolibdênico e DPPH. O extrato bruto e as frações acetato de etila e remanescente apresentaram atividade antioxidante, sendo que as duas últimas demonstraram maior atividade indicando que o conteúdo polifenólico pode ser responsável por esta atividade.

The polyphenolic content and the antioxidant activity of Euterpe edulis Martius fruit extract were assessed. This species is a Palm tree known as Juçara, native to Atlantic Forest and used for palm heart extraction. The process of palm heart extraction leads to the death of the plant since the latter has one single stem. The high demand has resulted in the depletion of this natural resource. Many species from Atlantic Forest can be used by means of sustainable management for the preservation and economic exploration by local communities. The fruit of Juçara palm can be one of the sustainable management alternatives for the Atlantic Forest natural resources. The antioxidant capacity of this fruit can justify its application as nutritional food. The fruit polyphenolic content was determined by the Folin-Ciocalteau method and the obtained results were: 10.31 ± 0.25%, 12.42 ± 0.89%, 12.75 ± 0.94% for crude extract, acetyl acetate and remaining fractions, respectively. The antioxidant capacity was determined through the phosphomolybdenium complex and DPPH methods. The crude extract and the acetyl acetate and remaining fractions showed antioxidant activity, and the latter two showed higher activity, indicating that polyphenolic content may be responsible for this activity.

Avaliação da atividade fitotóxica com enfoque alelopático do extrato das cascas de Celtis iguanaea (Jacq. ) Sargent Ulmaceae e purificação de dois triterpenos / Evaluation of the phytotoxic activity focused on the allelopathic effect of the extract from the bark of Celtis iguanaea (Jacq. ) Sargent Ulmaceae and purification of two terpenes

A espécie Celtis iguanaea (Jacq.) Sargent é popularmente conhecida como esporão de galo ou grão de galo. As folhas são indicadas pelo uso popular para o tratamento de dores no corpo e no peito, para reumatismo, asma, cólicas, má digestão e como diurético; as raízes são utilizadas para infecções urinárias e as cascas para a febre. O presente trabalho objetivou contribuir para o estudo fitoquímico e atividade fitotóxica com enfoque alelopático das cascas de Celtis iguanaea. O extrato etanólico foi submetido à partição com os solventes hexano, clorofórmio e acetato de etila. As substâncias friedelina e epifriedelinol (triterpenos) foram isoladas da fração hexano e identificadas por meio de métodos espectroscópicos de RMN de ¹H e 13C. O extrato bruto na concentração de 0,1 mg mL-1 causou inibição acentuada do hipocótilo em 34,97% e estimulou o crescimento da radícula em 29,64% de plântulas de Lactuca sativa. No ensaio de toxicidade frente à Artemia salina o extrato bruto e frações apresentaram uma CL50 superior a 1000 μg mL-1, indicando que o mesmo não possui efeito tóxico.

The species Celtis iguanaea (Jacq.) Sargent is popularly known as "esporão de galo" or "grão de galo". Its leaves are recommended by the popular use for the treatment of body and chest aches, as well as for rheumatism, asthma, cramps, indigestion and as diuretic; its roots are used for urinary infections and its bark for fever. This study aimed to contribute to the phytochemical investigation of the toxic activity focused on the allelopathic effect of the bark of Celtis iguanaea. The ethanol extract was subjected to solvent partition with hexane, chloroform and ethyl acetate. The substances friedelin and epifriedelinol (triterpenes) were isolated from the hexane fraction and identified by spectroscopic methods ¹H and 13C NMR. The crude extract at a concentration of 0.1 mg mL-1 caused marked inhibition of hypocotyl in 34.97% and stimulated radicle growth in 29.64% seedlings of Lactuca sativa. In the toxicity test against Artemia salina the crude extract and fractions showed an LC50 higher than 1000 μg mL-1, indicating that it has no toxic effect.

Functional polymorphisms in the interleukin-6 and serotonin transporter genes, and depression and fatigue induced by interferon-alpha and ribavirin treatment.

Depression and fatigue are frequent side effects of interferon-alpha (IFN-alpha) treatment, and there is compelling evidence that the inflammatory response system (including interleukin-6, IL-6) and the serotonergic system is important in the pathophysiology of such symptoms. Functional polymorphisms in the promoter region of the IL-6 gene (rs1800795) and serotonin transporter gene (5-HTTLPR) have been identified as regulating these systems. The present study aimed to determine if these polymorphisms were associated with the development of depression and fatigue during IFN-alpha and ribavirin treatment. Ninety-eight Caucasian patients receiving pegylated IFN-alpha and ribavirin treatment for chronic hepatitis C virus at King's College Hospital, London, and Emory University Hospital, Atlanta, participated in this prospective cohort study. Symptoms of depression and fatigue were measured before treatment and at weeks 4, 8, 12 and 24 during treatment. The 'low IL-6' synthesizing genotype (CC) was associated with significantly fewer symptoms of depression (effect size = 0.7 at week 24; F = 9.4, d.f. = 436, P = 0.002). The 'high transcription' serotonin transporter (5-HTT) genotype (LL) was also associated with significantly fewer symptoms of depression, but with a much smaller effect (effect size = 0.2 at week 24; F = 4.5, d.f. = 436, P = 0.03). Neither polymorphisms were associated with symptoms of fatigue (IL-6: F = 1.2, d.f. = 430, P = 0.2; 5-HTT: F = 0.5, d.f. = 430, P = 0.5). The smaller effects of the 5-HTT polymorphism on depression may be explained by an interaction between the genes (F = 5.0, d.f. = 434, P = 0.02): the 'protective' effect of the 5-HTTLPR polymorphism was evident only in the presence of the 'low IL-6' genotype (F = 5.4, d.f. = 64, P = 0.02), not in the presence of the 'high IL-6' genotype (F = 2.2, d.f. = 369, P = 0.1). The association between the IL-6 polymorphism and reduced risk of depressive symptoms confirms the role of the inflammatory response system in the pathophysiology of IFN-alpha-induced depression; in contrast, the effect of the 5-HTT gene was small and perhaps dependent on the status of the inflammatory response.

'Hypersynchronisation' by tissue velocity imaging in patients with cardiac amyloidosis.

OBJECTIVE: It is unknown if some patients with cardiac amyloidosis (CA) have mechanical dyssynchrony, as has been demonstrated in patients with ischaemic and dilated cardiomyopathies. The aim of this study was to assess mechanical dyssynchrony in patients with CA using tissue velocity imaging (TVI) and to define its usefulness for risk stratification. DESIGN AND PATIENTS: We included 121 patients with primary amyloidosis and 37 age-matched and sex-matched controls. Patients were divided into two groups: 60 with advanced-CA and 61 with no-advanced-CA, according to left ventricular (LV) wall thickness and diastolic dysfunction. Dyssynchrony assessment included: (1) atrioventricular dyssynchrony (dys), (2) interventricular dys, (3) intraventricular dys assessed longitudinally, using the standard deviation of time to systolic peak velocity (Ts-SD) of the 12 basal and mid level LV segments, and (4) intraventricular dys assessed radially, using the difference in radial Ts between mid anteroseptal and mid posterior segments. OUTCOME: Primary end-point was all-cause death. During a median follow-up of 13 months there were 35 events among patients. RESULTS: Contrary to the hypothesis, the intraventricular dys indices in advanced-CA patients were reduced compared to either the no-advanced-CA group or to controls (Ts-SD: 12.1 (9.0); 35.1 (18.6); 24.5 (14.1), respectively, p<0.001). This reduction was primarily the result of decreased ejection time (ET). Moreover, ET was the most significant predictor of survival (HR = 0.98, p<0.001). CONCLUSIONS: The regional timing of systolic motion measured by TVI was abnormally synchronised in the patients with advanced-CA. ET reduction plays a prominent part in this process and should be considered an essential parameter for assessment of patients with cardiac amyloidosis.

Increased risk of exacerbating gastrointestinal disease among elderly patients following treatment with calcium channel blockers.

BACKGROUND AND OBJECTIVE: Calcium channel blockers (CCBs) have been widely used for the treatment of hypertension and angina pectoris. It is presumed that CCBs decrease the lower esophageal sphincter pressure and as a result, the risk of gastrointestinal disease may be increased. Since the prevalence of gastrointestinal diseases has increased in elderly patients, it is possible that treatment with CCBs may have contributed to this increase. Therefore, we considered that the risk of exacerbating gastrointestinal disease among elderly patients by CCBs can be estimated by using the prescription ratio of antisecretory drugs as an outcome. METHOD: We hypothesized that patients who are prescribed CCBs would increase the use of antisecretory drugs involving H(2)-receptor antagonists and proton pump inhibitors (PPIs). From January 2001 to December 2005, a dynamic retrospective cohort study was performed at three community pharmacies in Nagasaki city, Japan, to assess the use of antisecretory drugs following treatment with CCBs among elderly patients. The correlation of initiation of antisecretory drugs treatment to maintenance therapy with PPIs was determined by the Cox proportional hazards model. RESULTS: The proposed study includes 303 patients prescribed CCBs and 258 controls. During the study period, 138 patients prescribed CCBs and 66 controls were initiated by giving antisecretory drugs; the hazard ratio was 1.40 (95% confidence interval 1.21-1.63). Eighty two patients taking CCBs and 32 controls were initiated by the maintenance therapy with PPIs; the hazard ratio was 1.48 (95% confidence interval 1.21-1.83). CONCLUSION: Patients who simultaneously initiated the use of antisecretory drugs with CCBs could not be found. Therefore, antisecretory drugs have not been used to prevent the gastrointestinal diseases caused by CCBs. The results obtained in this study suggest that the risk of gastrointestinal disease could be increased by long-term treatment with CCBs for elderly patients.

Engineering of the H2O2-binding pocket region of a recombinant manganese peroxidase to be resistant to H2O2.

The manganese peroxidase produced by Phanerochaete chrysosporium, which catalyzes the oxidation of Mn(2+) to Mn(3+), is easily inactivated by the hydrogen peroxide (H2O2) presented in the reaction. We attempted to increase H2O2 resistance by the conformational stabilization around the H2O2-binding pocket. Based on its structural model, engineering of oxidizable Met273 located near the pocket to a non-oxidizable Leu showed a great improvement. Furthermore, after treatment at 1 mM H2O2 where the wild-type is completely inactivated, full activity can be retained by engineering the Asn81, which might have conformational changes due to the environment of the pocket, to a non-bulky and non-oxidizable Ser.

The Purple leaf (Pl) locus of rice: the Pl(w) allele has a complex organization and includes two genes encoding basic helix-loop-helix proteins involved in anthocyanin biosynthesis.

The Purple leaf (Pl) locus of rice (Oryza sativa L.) affects regulation of anthocyanin biosynthesis in various plant tissues. The tissue-specific patterns of anthocyanin pigmentation, together with the syntenic relationship, indicate that the rice Pl locus may play a role in the anthocyanin pathway similar to the maize R/B loci. We isolated two cDNAs showing significant identity to the basic helix-loop-helix (bHLH) proteins found in the maize R gene family. OSB1 appeared to be allelic to the previously isolated R homologue, Ra1, but showed a striking difference at the C-terminus because of a 2-bp deletion. Characterization of the corresponding genomic region revealed that the sequence identical to a 5'-portion of OSB2 existed approximately 10-kb downstream of the OSB1 coding region. OSB2 lacks a conserved C-terminal domain. Restriction fragment length polymorphism analyses using an F(2) population indicate that both genes co-segregate with the purple leaf phenotype. A transient complementation assay showed that the anthocyanin pathway is inducible by OSB1 or OSB2. These results suggest that the Pl(w) allele may be complex and composed of at least two genes encoding bHLH proteins.

Differences between the tolerance characteristics of two anticonvulsant benzodiazepines in the amygdaloid-kindled rat.

The characteristics of the development of tolerance to the anticonvulsant effects of chronic treatment by dipotassium clorazepate and diazepam using amygdaloid-kindled rats were investigated. Dipotassium clorazepate (5 mg/kg) or diazepam (5 mg/kg) were intraperitoneally administered for 10 consecutive days. Tolerance to the anticonvulsant effect of dipotassium clorazepate developed in seizure stage on day 6, after-discharge duration on day 7 and seizure latency on day 4. In contrast, tolerance to the effects of diazepam developed more rapidly in seizure stage on day 4, after-discharge duration on day 4 and seizure latency on day 3. Thus tolerance to the anticonvulsive effect of dipotassium clorazepate developed relatively slower than that to diazepam. All rats had stage 5 convulsions 24 hr after cessation of the administration of dipotassium clorazepate and diazepam. Concomitant determinations of plasma concentrations of the main metabolite of dipotassium clorazepate and diazepam, desmethyldiazepam, showed no statistical difference during treatment, suggesting that the developed tolerance was not metabolic but functional.

[A case of lung cancer in which FDG-PET was useful for preoperative diagnosis].

The patient was 48-year-old woman. The patient was referred to our hospital due to detection of an abnormal shadow on a chest X-ray image during a health checkup. The results of a CT scan morphologically suggested a benign tumor, but a diagnosis could not be made even after bronchoscopic cytology. FDG-PET revealed accumulation at the tumor site suggestive of malignancy, and an operation was performed. Prompt pathological examination showed that the tumor was adenocarcinoma, and right lower lobectomy and lymphnode dissection were performed. FDG-PET is thought to be useful for diagnosis in cases of lung tumor in which a preoperative diagnosis can not be established.

Assessment of coronary flow velocity with transthoracic Doppler echocardiography during dobutamine stress echocardiography.

OBJECTIVES: The purpose of this study was to evaluate the feasibility of measuring coronary flow velocity (CFV) by transthoracic Doppler echocardiography (TTDE) in the left anterior descending coronary artery (LAD) during contrast-enhanced dobutamine stress echocardiography (DSE). We also assessed the value of TTDE for detecting stress-induced myocardial ischemia in the LAD territory. BACKGROUND: Noninvasive assessment of both CFV and wall motion during DSE would enhance the diagnostic accuracy of DSE. METHODS: One hundred forty-four consecutive patients underwent CFV recording in the distal LAD by TTDE during contrast-enhanced DSE. Regional wall motion score index (WMSI) in the LAD territory and CFV ratio at peak stress (CFV ratio peak), defined as a ratio of CFV at peak stress to basal CFV, were obtained. RESULTS: Coronary flow velocity was successfully recorded in 129 patients (90%) at baseline and during dobutamine infusion. Mean value of CFV ratio peak was 2.39 +/- 0.83 (range: 0.84 to 4.40). There was good correlation between WMSI at peak stress and CFV ratio peak (r = 0.62, p < 0.001). Coronary flow velocity ratio peak was significantly lower in patients who developed stress-induced wall motion abnormality (WMA) in the LAD territory than it was in those patients without WMA (1.51 +/- 0.51 vs. 2.76 +/- 0.65, p < 0.001). A CFV ratio peak <2.1 had a sensitivity of 92% and a specificity of 86% for detecting the presence of stress-induced WMA. CONCLUSIONS: Assessment of CFV in the distal LAD during DSE is feasible in the majority of cases and provides a CFV ratio for detecting stress-induced myocardial ischemia in the LAD territory.

Unusual occipital condyle fracture with multiple nerve palsies and Wallenberg syndrome.

A 52-year-old male presented with an extremely rare fracture of the occipital condyle involving the jugular foramen with marked medial rostrad displacement of the fragments. He had ipsilateral VII through XII nerves palsies and Wallenberg syndrome. Conservative treatment did not improve the cranial nerve palsies. A high-resolution CT-scan is essential to visualize these fractures.

Characteristics of epilepsy in severely mentally retarded individuals.

In order to clarify the characteristics of epilepsy in patients with severe mentally retarded (SMR) subjects, we analyzed 52 SMR subjects with epilepsy from the institute for SMR subjects at Kikuchi National Hospital, Kumamoto, Japan. A total of 61.5% patients had uncontrolled seizures which were resistant to treatment. The most common combinations of seizure types in those not responding to conventional anticonvulsants were generalized tonic-clonic seizures (GTCS) with tonic seizure and GTCS with atypical absence. Their clinical features were characterized by spastic paralysis associated with a slower background electroencephalogram and abnormal computed tomography scans of the head, suggesting the involvement of cortical damage. These findings suggest that a large proportion of epilepsy in SMR subjects does not respond to treatment and that the severity of organic brain damage may therefore affect the natural course of epilepsy in such patients.

Effect of dipotassium clorazepate on amygdaloid-kindling and comparison between amygdaloid- and hippocampal-kindled seizures in rats.

We examined the effect of dipotassium clorazepate (7-chloro-1, 3-dihydro-2-oxo-5-phenyl-1H-1, 4-benzodiazepine-3-carboxylate potassium hydroxide), an antianxiety drug, on amygdaloid kindling and compared its effects for 7 successive days on amygdaloid- versus hippocampal-kindled seizures, using the rat kindling model of epilepsy. Dipotassium clorazepate at 5 mg/kg significantly delayed amygdaloid kindling. The contralateral cortical after-discharge duration in the dipotassium clorazepate-treated group was significantly shorter than the after-discharge duration in the amygdala in the first seven stimulations, whereas it was significantly shorter only in the first three stimulations in the control group, indicating that dipotassium clorazepate suppressed the spread of seizure activity from focus to contralateral cortex. Dipotassium clorazepate suppressed amygdaloid-kindled seizures at 2 and 5 mg/kg, while 1 mg/kg or more suppressed hippocampal-kindled seizures. Thus, differences in effective dosages in both amygdaloid- and hippocampal-kindled seizures may suggest a difference in the neuronal mechanisms involved in this kindling.

Changes in the specificity of antibodies by site-specific mutagenesis followed by random mutagenesis.

The specificity for 11-deoxycortisol (11-DOC) of a monoclonal antibody (mAb), designated SCET, was changed to specificity for cortisol (CS) by site-specific mutagenesis followed by random mutagenesis. The Fab form of SCET was expressed on the surface of a phage. During the first step, mutations were introduced at 14 amino acid positions in three complementarity-determining regions (CDRs) of the VH domain that seemed likely to form the steroid-binding pocket. A clone, DcC16, was isolated from the resultant library with multiple mutations and this clone was shown to have CS-binding activity but also to retain high 11-DOC-binding activity. During the second step, mutations were introduced randomly into the entire VH-coding region of the DcC16 clone by an error-prone polymerase chain reaction, and CS-specific mutant antibodies were selected in the presence of 11-DOC as a competitor. Three representative clones were analyzed with the BIAcore instrument, and each revealed a large increase in the binding constant for CS and a decrease in that for 11-DOC. Structural models, constructed by computer simulation, indicated the probable molecular basis for these changes in specificity.

Host immune responses to ribosome, ribosomal proteins, and RNA from Mycobacterium bovis bacille de Calmette-Gúerin.

The ribosomes from BCG strongly induced delayed type hypersensitivity (DTH) skin reactions in guinea pigs immunized with live BCG or heat killed Mycobacterium tuberculosis H37Rv, and also induced lymphocyte proliferative response in mice immunized with ribosomes. In contrast, neither ribosomal proteins nor RNA alone induced both DTH skin reactions and lymphocyte proliferative responses. Particle form consisted of ribosomal proteins and RNAs might be absolutely required for the activation of immune responses.

Isolation of a protease-deficient mutant of Bacillus brevis and efficient secretion of a fungal protein disulfide isomerase by the mutant.

The efficient production of a thermostable protein disulfide isomerase (PDI) was successfully achieved using the newly isolated protease-deficient mutant, Bacillus brevis 31-OK. Extracellular protease (exoprotease) activity was about a quarter of that in the parent, and the mutant was deficient in at least one of the major exoproteases. The cDNA encoding the fungal PDI was inserted downstream of the signal peptide-encoding region in an expression-secretion vector for B. brevis. Efficient production of PDI was feasible using B. brevis 31-OK as a host and modified signal sequences composed of three leucine residues inserted in the hydrophobic region of the MWP (middle wall protein) signal sequence. The maximal secretion of PDI into the culture medium was 1.1 g/l, which is about twice that by the parent strain and fifty times greater than the amount of rat and murine PDIs produced by Escherichia coli. The enzymatic properties such as the specific activity and thermal stability of the recombinant PDI are similar to those of natural PDI derived from Humicola insolens mycelia. B. brevis 31-OK was able to maintain its exoprotease activity at a low level throughout the cultivation and is considered to be useful host for production of a protease-sensitive protein and for increase of protein productivity due to stable accumulation.

RFLP mapping of BaYMV resistance gene rym3 in barley (Hordeum vulgare).

The rym3 (formerly designated ym3) gene conferring resistance to barley yellow mosaic virus (BaYMV) is effective against all strains of the virus but up to now has not been mapped to any chromosome. We performed a linkage analysis, using DNA extracted from individually harvested mature leaves of 153 F(2) plants derived from a cross between BaYMV-resistant cv 'Ishuku Shirazu' carrying rym3 and susceptible cv 'Ko A'. Additionally, the F(3) lines derived from F(2) plants were grown in the BaYMV-infested field and examined for their reaction to BaYMV. Our results indicated that rym3 is located on the short arm of chromosome 5H and flanked by RFLP markers MWG28and ABG705A at distances of 7.2 and 11.7 cM, respectively. The chromosomal configuration estimated by DNA markers around rym3 and the utilization of these molecular markers for pyramiding with the BaYMV resistance genes in barley breeding programs are discussed.

Anticonvulsant effects of dipotassium clorazepate on hippocampal kindled seizures in rats.

We examined the anticonvulsant properties of dipotassium clorazepate (DC) against hippocampal kindled seizures in rats. Adult male Wistar rats were subjected to kindling 1 week after the implantation of electrodes. After five stage 5 seizures were induced, the generalized convulsion triggering threshold (GST) was determined. Dipotassium clorazepate was administered intraperitoneally in rats that showed two stable stage 5 seizures induced at the GST current intensity. Dipotassium clorazepate at doses of 1 mg/kg or more produced an anticonvulsant effect, but did not readily suppress limbic seizures. Dipotassium clorazepate did not completely suppress after-discharges (AD) even at the highest dose, which was 5 mg/kg. Moreover, raised stimulus intensity failed to affect its efficacy as an anticonvulsant. The results of the present study suggest that DC has a modest anticonvulsant potency. It is reasonable to assume that its anticonvulsant efficacy is primarily due to attenuation of AD propagation rather than the raising of the seizure triggering threshold at the kindling focus.