Dried, ground banana plant leaves (Musa spp.) for the control of Haemonchus contortus and Trichostrongylus colubriformis infections in sheep.

To evaluate the anthelmintic effect of Musa spp. leaves, 12 animals were artificially infected with Haemonchus contortus, and another 12 animals were infected with Trichostrongylus colubriformis. Then, both treatment groups were offered 400 g of dried ground banana plant leaves, and the control animals were offered only 1000 g of coast cross hay. During the trials, the animals received weekly physical examinations. The methods used to evaluate the efficiency of this treatment were packed cell volume, total plasma protein and faecal egg counts, and egg hatchability tests were performed on days -2, +3, +6, +9, +13 and +15. Coproculture tests were performed on day -2 to confirm monospecific infections. In the FEC and EHT, a statistically significant difference (0.04, 0.005; p < 0.05) was noted for T. colubriformis. There were no statistically significant differences (p > 0.05) for Haemochus contortus group in all tests. Our results confirmed previous findings suggesting that dried ground banana plant leaves possess anthelmintic activity.

Preliminary results after Single-Incision Laparoscopic Colorectal Surgery.

OBJECTIVE: Single-incision laparoscopic surgery (SILS) represents the recent advance in laparoscopic surgery claiming to be less invasive than conventional laparoscopic surgery. This study investigates the feasibility and safety of the procedure in colorectal surgery and reports the experiences in our center. METHODS: We retrospectively evaluated 41 consecutive patients surgically treated in our institution (February 2011-April 2013). The patient characteristics were evaluated for: gender, age, body mass index and ASA-score. Data included: indication, procedure, intraoperative complications, operation time, number of lymph nodes, postoperative complications, length of stay (LOS), morbidity and cosmesis. RESULTS: SILS was performed in 41 patients including 9 patients with colorectal cancer resection. We performed 3 ileocaecal resections, 11 right hemicolectomies, 7 sigmoidectomies and 20 rectosigmoidectomies. The operation time ranged from 45-210 min (median 123 min). Number of lymph nodes identified, ranged from 1-37 (median n=8). Six post-operative complications (14%) occurred: 1 gastroparesis, 1 subobstruction, 1 anastomotic leak and 3 patients needed a blood transfusion postoperatively. Median LOS was 6 days (range 4-21 days). One delayed complication (2,4%) occurred (eventration). None of the patients died. All patients had satisfactory cosmetic results. CONCLUSIONS: With the proviso that the study population was limited in size, SILS is feasible and is a save procedure in colorectal surgery and the procedure has an obvious cosmetic benefit. The results are comparable to other international reports. Still the procedure should be restricted to selected patients and performed by experienced surgeons. Additional prospective studies are essential to define the further benefit.

Prognostic value of power doppler ultrasonography for equine superficial digital flexor tendon injury in thoroughbred racehorses.

The potential value of hypervascularity detected with power Doppler ultrasonography (PDU) within equine superficial digital flexor tendon (SDFT) as a prognostic factor of SDFT injury is not clear. The purpose of this study was to test the hypothesis that hypervascularity within SDFT is one of the risk factors for subsequent severe SDFT injury and to evaluate the prognostic value. A prospective cohort study of 97 Thoroughbred racehorses without any clinical signs of SDFT injury was conducted. Six variables of age, body weight, sex, the cross-sectional area of SDFT, PDU signal within SDFT and experience of steeplechase were assessed for the possibility of risk factors of subsequent SDFT injury in follow-up period of 1 year. Multivariable logistic regression analyses were used for assessment of the odds ratios (ORs) and 95 % confidence intervals (CIs) of SDFT injury. Multivariable logistic regression analysis revealed that the PDU signal within SDFT was a risk factor for the development of SDFT injury in follow-up period (P = 0.017). The adjusted OR of SDFT injury was significantly higher in PDU positive group than in PDU negative group (OR 3.17, 95 % CIs 1.20-8.35). Although further studies are required, these results would be useful for early detection and/or prevention of development for clinical severe SDFT injury.

Association between N-methyl-D-aspartate Receptor Subunit 2B Gene Polymorphisms and Personality Traits in a Young Japanese Population.

OBJECTIVE: The N-methyl-D-aspartate receptor subunit 2B (GluN2B) is involved in regulation of anxiety and depression and nervous activity in the brain. Single nucleotide polymorphisms of the GluN2B gene (GRIN2B) are associated with human mental function and behaviour. We investigated whether four GRIN2B polymorphisms (rs7301328, rs1806201, rs1805247, and rs1805502) affect characterisation of personality traits. METHODS: In 248 young people, GRIN2B polymorphisms were analysed, and personality traits were assessed using the Neuroticism Extraversion Openness-Five Factor Inventory (NEO-FFI) and State-Trait Anxiety Inventory (STAI). RESULTS: There was no main effect of the GRIN2B polymorphisms on the NEO-FFI and STAI dimension scores. Interaction between polymorphism and sex was found in rs1805247 (p = 0.034) and rs1805502 (p = 0.040) in terms of the conscientiousness score of the NEO-FFI. However, post hoc simple main effect analysis showed no significant effect. The preliminary haplotype analysis indicated that haplotype CTT (rs1806201-rs1805247-rs1805502) in the haplotype block was associated with the extraversion score of the NEO-FFI in female participants (p = 0.044), but the significance was lost on correction for multiple testing. CONCLUSION: There was no significant association between selected GRIN2B polymorphisms and personality traits, but this may be due to low statistical power. Further studies involving a larger study population are needed to clarify this.

A retrospective cohort study investigating risk factors for the failure of Thoroughbred racehorses to return to racing after superficial digital flexor tendon injury.

A retrospective cohort study was conducted to investigate risk factors for the failure of Thoroughbred racehorses to return to racing after an injury of the superficial digital flexor tendon (SDFT). Successful return was defined as the completion of five or more races after SDFT injury. The official Japan Racing Association (JRA) medical records of racehorses with a core-type SDFT injury were reviewed for clinical variables related to the characteristics of the horse and the severity of SDFT injuries at the time of diagnosis. Data on racing outcomes were obtained from the official JRA racing database. Risk factors were screened using univariable logistic regression and subsequent multivariable model building. Forty-nine of 346 (14.2%) horses successfully returned to racing after SDFT injuries. Multivariable model building revealed that an increase in the total number of injured zones (defined as the total number of zones in which the injured hypoechoic area was observed at the time of ultrasonographic diagnosis of SDFT injury) was associated with an increased risk of failure to return to racing after SDFT injury. Horse characteristics, such as age, body mass and sex, were not associated with a successful return to racing. In the rehabilitation of cases with larger (longer) lesions, more effective and careful medical management may be needed for an improvement in the athletic outcomes.

Cytolytic activity of liposomes containing stearylamine.

In order to develop the cytotoxic liposome, the cytolytic effect of polycationic liposome was examined. Upon incubation of the stearylamine-containing liposome (stearylamine-liposome) with rabbit erythrocyte, a significant extent of hemolysis was observed. Hemolytic activity of the liposome depends on the amount of stearylamine in the liposome membrane. The plots of the initial rate of hemolysis versus the concentration of stearylamine-liposome showed a sigmoidal curve, suggesting that stearylamine-liposomes act cooperatively on the erythrocyte membrane. Hemolytic activity of stearylamine-liposome was markedly influenced by the composition of hydrocarbon chains of the phospholipids in the liposome membrane, suggesting that the membrane fluidity of stearylamine-liposome is important to evoke the hemolysis. Since the liposomes containing acidic phospholipids inhibited markedly the stearylamine-liposome-caused hemolysis, it is likely that the primary target of stearylamine-liposome is the negatively charged component(s) such as acidic phospholipids on the erythrocyte membrane. Furthermore, stearylamine-liposome induced the release of the intravesicular contents from the liposome made of acidic phospholipids but not from the liposome made of phosphatidylcholine only. These results suggest that stearylamine-liposome interacted with the negative charges of the erythrocyte membrane and eventually damaged the cell. Erythrocytes from rabbit, horse and guinea pig are highly susceptible to stearylamine-liposome but those from man, sheep, cow and chicken are less so.

Separation of gate- and channel-forming domains in the pore-forming protein of the outer membrane of Pseudomonas aeruginosa.

The domains of the pore-forming protein responsible for the gate and channel formations were separated and identified in the outer membrane of Pseudomonas aeruginosa. The proteolytic cleavage of the 46K channel protein, protein D2, yielded two major domains with apparent M(r) of 27K and 19K. We identified the 27K polypeptide to be the channel-forming domain by an in vitro permeability assay. The channel size of purified 27K domain was indistinguishable from that of native protein D2. Degradation of the 19K domain into small subfragments increases the channel activity about ten times suggesting that the 19K polypeptide forms the gate or cap.

Protein D2 porin of the Pseudomonas aeruginosa outer membrane bears the protease activity.

We report here our discovery that protein D2 of the outer membrane of Pseudomonas aeruginosa is a novel porin bearing protease activity. Homogeneously purified protein D2 hydrolyzed several synthetic peptides according to the Michaelis-Menten kinetics. A specific serine protease inhibitor, diisopropyl fluorophosphate (DFP), inactivated the protease activity and [3H]DFP covalently labeled protein D2. We tested the effect of two monoclonal antibodies raised against protein D2 on the protease activity. One antibody lowered the protease activity to about 20%, while the other enhanced it to about 300% of that without antibody. In addition, the fractions derived from the outer membrane of the protein D2-deficient mutants showed negligible protease activity, whereas similarly fractionated outer membrane proteins of the protein D2-positive parent strain showed strong protease activity.

Blood coagulation factor IX-binding protein from the venom of Trimeresurus flavoviridis: purification and characterization.

The coagulation factor IX/factor X-binding protein (IX/X-bp) from the venom of Trimeresurus flavoviridis is a heterogeneous two-chain protein, and the structure of each chain is similar to that of the carbohydrate-recognition domain of C-type lectins, such as asialoglycoprotein receptors, pancreatic stone protein, and the Fc epsilon receptor for immunoglobulin E. Analysis of the binding properties of IX/X-bp revealed that it binds to the gamma-carboxyglutamic acid (Gla)-containing domains of factors IX and X [Atoda, H. et al. (1994) Eur. J. Biochem. 224, 703-708]. In the present study, we isolated another anticoagulant protein that binds to factor IX but is not to factor X. This protein, designated IX-bp, inhibited factor IXa-induced clotting but not factor Xa-induced clotting, whereas IX/X-bp inhibits both. The concentration of IX-bp for half-maximal binding to solid-phase bovine factor IX was 0.4 nM whereas IX-bp did not bind to factor X even at 40 nM. The binding of IX-bp to solid-phase factor IX was inhibited by the addition of Gla-domain peptide of factor IX, indicating that IX-bp binds to the Gla-domain region of factor IX. IX-bp had two Ca(2+)-binding sites with different affinities for Ca2+ ions. At pH 7.5, the apparent Kd values for these sites were 14 and 130 microM, respectively. IX-bp was a two-chain protein (27.5-kDa band before reduction and 16.8- and 15.7-kDa bands after reduction on SDS-PAGE) and it reacted with immunoglobulin G against IX/X-bp. The complete amino acid sequence of IX-bp was determined. The 16.8-kDa chain (A chain) of IX-bp consisted of 129 residues, of which 19 were different from those in the A chain of IX/X-bp (129 residues). The sequence of the 15.7-kDa chain (B chain) was identical to that of the B chain of IX/X-bp (123 residues). We conclude that IX-bp is a protein that is structurally similar to but functionally different from IX/X-bp. The difference of binding specificity between IX-bp and IX/X-bp presumably arises from the sequence differences in the A chains.

cDNA cloning of a heterogeneous two-chain anticoagulant protein IX-bp.

IX-bp and IX/X-bp are heterogeneous two-chain anticoagulant proteins isolated from the venom of habu snake, Trimeresurus flavoviridis. The amino acid sequence of one (B chain) of their two chains is identical. We recently reported the cloning of cDNA encoding the B chain and that of the A chain of IX/X-bp. Here, we report the isolation and characterization of cDNA clones encoding the A chain of IX-bp. The 697-bp sequence showed a putative ORF capable of encoding a 152-amino-acid protein containing a 23-amino-acid signal peptide. The overall amino acid sequence identity between the pre-A chain of IX-bp and that of IX/X-bp is 84%, whereas the cDNA sequence identity of the two ORFs is 91% indicating that habu snake acquired the variety of venom proteins by efficient mutations of antonymous sites.

Trypanocidal activity of the stearylamine-bearing liposome in vitro.

Liposome made of stearylamine and phosphatidylcholine showed the trypanocidal activity in vitro. Cytotoxicity of the liposome against Trypanosoma cruzi appeared to be the strongest in trypomastigotes followed by amastigotes and epimastigotes. Lysis of the human erythrocyte was undetectably low under the conditions that the liposome kills more than 95% of trypomastigotes. The liposome seems to damage the plasma membrane.

Mechanism of relaxant response to papaverine on the smooth muscle of non-pregnant rat uterus.

In the present paper, we examined a mechanism of the papaverine induced relaxation in the smooth muscle of non-pregnant rat uterus. The hyperosmotic 65 mM KCl (H-65K+)-or oxytocin-induced contraction in the uterus was inhibited by an addition of papaverine in a concentration-dependent manner. Papaverine did not increase both cAMP and cGMP contents in the uterus in the presence of H-65K+ or oxytocin. In fura 2 loaded muscles, papaverine did not affect an increase of [Ca2+]i level by high K+ or oxytocin. In permeabilized muscles, papaverine had no effect on the Ca2+-induced contraction. H-65K+ and oxytocin increased the rate of oxygen consumption 1.8 and 1.5 times higher than that in the resting condition, respectively. The increase of oxygen consumption in the H-65K+ or oxytocin was significantly inhibited by papaverine (1-100 microM). These results suggested that papaverine inhibits smooth muscle contraction mainly by inhibition of mitochondrial respiration in rat uterus as well as guinea pig ileum, which shows a highly spontaneous activity and a highly metabolic dependency of a contraction.

Characteristics of Japanese alcoholics with inactive aldehyde dehydrogenase: clinical features of alcoholics with ALDH2*2.

Abstract In a person with inactive ALDH2 (ALDH2*2) the blood aldehyde concentration tends to rise faster and higher and there are flushing responses which are considered to be a restraint against excessive alcohol drinking. The subjects in this study comprised 71 Japanese alcoholics. Psychiatrists interviewed the patients concerning the clinical features. Alcoholics homozygous (n = 59) for ALDH2*1/ALDH2*1 (Group I) and those heterozygous (n = 12) for ALDH2*1/ALDH2*2 (Group II) were compared. Group II alcoholics included significantly more cases of guilt or personality disorder. These findings indicate that alcoholics with the ALDH2*2 genotype showed generally typical clinical features.

Protection of Toxoplasma gondii-infected mice by stearylamine-bearing liposomes.

The cytotoxic activity of stearylamine-bearing liposomes against Toxoplasma gondii (RH strain) was examined. When tachyzoites were treated in vitro with liposomes consisting of 20 mol% stearylamine and 80 mol% phosphatidylcholine (130 micrograms/ml total lipids), more than 95% of the parasites were killed within 90 min. Intraperitoneal injection of 10 mg of 30 mol% stearylamine/70 mol% phosphatidylcholine-liposomes in mice shortly before or after T. gondii challenge afforded protection from death for more than 30 days to 70-80% of the treated mice, whereas all untreated mice succumbed within 9 days. The liposome-injected mice that survived remained symptom-free and behaved normally.

[Properties of extracellular products produced by group A streptococci isolated from patient with streptococcal toxic shock syndrome].

Extracellular products of group A streptococci isolated from patients with streptococcal toxic shock syndrome (STSS) examined. The outline of discussion of the 4 products are as follows; Products were streptolysin O (SLO), proteinase, erythrogenic toxin and DNase. SLO showed a large amount of products more than proteinase, erythrogenic toxin and DNase. It should not be concluded that there is a large amount of SLO in the isolated strains from patients with STSS. SLO produced by group A streptococci isolated from the patient with STSS has an isoelectric point (pI) of 6.0 and a molecular weight of 64,000 and shows a hemolytic activity in the presence of 2 mercaptoethanol (2-ME). Furthermore, the hemolytic activities of all components were inhibited by gamma-globulin and cholesterol. These results indicated that all component with hemolytic activity are SLO. These data suggest that STSS does not make up a single syndrome but, rather, that the multiple STSS clinical criteria probably reflects a different characteristic of individual S. pyogenes isolates.

[Type specific protein antigen of group B streptococci [II]. Diversity of the c alpha protein antigen].

The c protein antigen of group B streptococci has been reported to consist of at least two acid extractable antigens, c alpha and c beta. We previously reported that strain FZ301 contain the c alpha and novel protein antigen, named as S antigen by Denka Seiken, and the c alpha antigen of this strain exhibit a more complicated structure. In this report described the analysis of the diversity of the c alpha antigen by immunoelectrophoresis and immunodiffusion. Hot acid extract from strain A909 produced a single precipitin band against the anti-121424c alpha serum but the 121424 extract produced double precipitin bands by immunoelectrophoresis. These c alpha antigens showed anodic migration, but the 121424c alpha antigens tended to migrate slower than the A909c alpha antigen. Moreover, the anti-A909c alpha serum produced a single precipitin band when tested after electrophoresis of the 121424 extract. When examined by immunodiffusion, the anti-121424c alpha serum produced two distinct precipitin bands with the 121424 and FZ301 extract, and these were reactions of identity. However, the A909 extract produced only one band which joined one of the two bands produced with the 121424 extract. These results shown that the strain 121424 contain c alpha and S antigen as well as strain FZ301. The precipitin band of the c alpha antigen in the A909 extract against the anti-A909 serum formed the spur with the reaction of the c alpha antigen in the 121424 or FZ301 extract on immunodiffusion. Moreover, the c alpha antibody in the anti-A909 serum failed to removed completely by repeated adsorption with strain 121424.(ABSTRACT TRUNCATED AT 250 WORDS)

[Properties of a hemolysin produced by group B streptococci].

The hemolysin produced by group B streptococci (GBH) has an isoelectric point (pI) of 5.8 and it shows a hemolytic activity in the absence of 2-mercaptoethanol (2-ME). The hemolytic activities of GBH were compared to that of streptolysin O (SLO) and streptolysin S (SLS). These hemolysins differed with respect to the binding and release of hemoglobin (Hb). GBH was bound to phospholipids on the membranes of target erythrocytes, followed by the gentle release of K+ and slow Hb release without lag time. Incontrast SLO released Hb as rapidly as K+. GBH induced hemolysis was inhibited by the addition of 30 mM raffinose. These results indicate that the effective diameter of the pores formed by GBH was about 1.1 nm. GBH showed a lower hemolytic efficiency than SLO, reflecting the fact that these hemolysins destroy erythrocytes by a different mechanism. Intracellular K+ and Hb were released at a different rate in GBH treated cells, indicating that a colloid-osmotic process is involved in the lytic mechanism.

[Properties of a hemolysin produced by group C streptococci isolated from hemolytic streptococcal infection in formosan squirrels].

The hemolysin produced by group C streptococci (GCH) has an isoelectric point (pI) of 5.6 and a molecular weight of 32,000 and shows hemolytic activity in the absence of 2-mercaptoethanol (2-ME). The hemolytic activity of GCH was compared to that of streptolysin S (SLS). These hemolysins did not differ with respect to the binding and release of hemoglobin (Hb). GCH was bound to phospholipids on the membranes of target erythrocytes, followed by the rapid released of K+ and slow Hb release after a relatively long lag time. GCH showed a lower hemolytic efficiency than SLS, reflecting the fact that these hemolysins destroy erythrocytes by slightly different mechanisms.

[Properties of extracellular products produced by group A streptococci isolated from patients with streptococcal toxic shock syndrome].

Extracellular products of group A streptococci isolated from patients with streptococcal toxic shock syndrome (STSS) were examined. The outline of the discussion of the 3 products are as follows; streptolysin O (SLO), proteinase and erythrogenic toxin. SLO and proteinase showed a relatively large amount of products more than erythrogenic toxin. SLO produced by group A streptococci isolated from the patient with STSS had an isoelectric point (pI) of 6.0 and a molecular weight of 64,000 and showed hemolytic activity in the presence of 2-mercaptoethanol (2-ME). Furthermore, the hemolytic activities of all components were inhibited by gamma-globulin and cholesterol. Proteinase had pIs of 8.7 and 8.9, and a molecular weight of 21,000. These data suggest that STSS clinical criteria probably reflects a characteristic of a large amount of products of individual S. pyogenes isolates.

[Relationship between alcoholism and HTR2 MspI polymorphism].

We investigated the human serotonin receptor, HTR2 genotype among 73 alcoholics. We found that there might not be a significant difference between alcoholics and controls in the frequency of HTR2 C1/C2 gene (MspI polymorphism). This result suggested that the HTR2 C1/C2 gene might not be associated with the risk factor for developing alcohol dependence. Further studies are required to determine whether or not a novel serotonin receptor polymorphism reflects pathogenesis of alcoholism.