Stable isotopes to study sulfur amino acid utilization in broilers.

Nutritionists have been discussing whether the dietary supplementation of cyst(e)ine is required as a part of the dietary methionine (Met) in the total sulfur amino acid (TSAA) requirement to achieve optimum performance in broilers. Part of Met is converted to cysteine (Cys) to meet the Cys requirement, especially for feather growth. The TSAA requirement has been determined by using graded levels of free Met in the diet, without supplementation of free cyst(e)ine. It has also been argued that the Met to Cys ratio (Met : Cys) changes with age and even with different Met sources. The objective of this study was to evaluate the two sources of Met, while determining the proportion of Met and Cys in total dietary TSAA that optimize the performance of broilers. A performance assay was carried out in a factorial arrangement (5 × 2) using 1080 broilers from 42 to 56 days of age fed diets having different dietary proportions of Met and Cys (44 : 56, 46 : 54, 48 : 52, 50 : 50 or 52 : 48) while maintaining the same dietary TSAA in the diets. Two synthetic Met sources (dl-Met or l-Met) were used for each of the diets with different dietary Met : Cys ratios. Twenty-one broilers of the same age were fed the diets 44 : 56, 48 : 52 and 52 : 48 by supplementing the diet with L-(15N) Met or L-(15N2) Cystine to study the metabolism of TSAA. No differences were observed between Met sources for feed intake, BW gain and feed conversion ratio (FCR; P > 0.05); however, FCR was numerically improved at 50 : 50 Met : Cys. Regarding TSAA utilization, the conversion of Met to Cys increased with increase in Met : Cys ratios, but the concentration of Met intermediates decreased. Broiler chickens responded to different dietary proportions of sulfur amino acids by altering their sulfur amino acid metabolism, and diets containing 50 : 50 Met : Cys is recommended for broilers of age 42 to 56 days.

Germinação assimbiótica e desenvolvimento de Dendrobium nobile Lindl. sob efeito de reguladores vegetais no tratamento pré-germinativo / Asymbiotic germination and development of Dendrobium nobile Lindl. under the effect of plant growth regulators in pre-germinative treatment

O presente trabalho teve como objetivo estudar a influência dos reguladores vegetais BAP e GA3 como tratamentos pré-germinativos no processo de germinação e desenvolvimento inicial de plântulas de Dendrobium nobile, espécie importante pelas propriedades farmacológicas como anti-oxidante, vasodilatadora e até mesmo anti-cancerígena, além do valor ornamental. Os tratamentos pré-germinativos consistiram de BAP e GA3, separadamente, nas concentrações de 0,0; 1,0; 2,0 e 5,0 mg L-1. Após seis meses da semeadura in vitro e manutenção em câmara de germinação e de crescimento com temperatura e foto-período controlados (12 horas e 23ºC ± 2), foram avaliados os parâmetros número de sementes germinadas, porcentagem de germinação, massa fresca e altura das plântulas, diâmetro e número de pseudobulbos, número de folhas, número de raízes, e o comprimento da maior raiz. O delineamento experimental foi inteiramente casualizado. Todas as variáveis foram submetidas à análise de variância e de regressão, quando significativas. As sementes de D. nobile germinaram melhor na ausência de reguladores vegetais e os tratamentos com BAP ou GA3 na embebição das sementes pouco beneficiaram o desenvolvimento in vitro de D. nobile.

The present study aimed to investigate the influence of plant growth regulators BAP and GA3 as pre-germinative treatment in the process of germination and initial development of seedlings of Dendrobium nobile, a species important for its pharmacological properties like antioxidant, vasodilator and even anticancer, besides its ornamental value. Pre-germinative treatments consisted of BAP and GA3, separately, at the concentrations of 0.0; 1.0; 2.0 and 5.0 mg L-1. At six months after in vitro sowing and maintenance in a germination and growth chamber with controlled temperature and photoperiod (12 hours and 23ºC ± 2), the following parameters were evaluated: number of germinated seeds, percentage of germination, fresh mass and height of seedlings, number and diameter of pseudo-bulbs, number of leaves, number of roots, and length of the largest root. The experimental design was completely randomized. All variables underwent analysis of variance and regression analysis when significant. D. nobile seeds presented better germination in the absence of plant growth regulators and the treatments with BAP or GA3 in seed imbibition little benefited D. nobile in vitro development.

In vitro evaluation of cashew nut shell liquid as a methane-inhibiting and propionate-enhancing agent for ruminants.

Cashew nut shell liquid (CNSL) containing antibacterial phenolic compounds was evaluated for its potency as a feed additive for ruminants. In experiment 1, ruminal responses to CNSL supplementation were assessed using a batch culture system. Rumen fluid from cattle was diluted with artificial saliva and incubated for 18h in a batch culture with a mixed diet containing a 30:70 hay:concentrate diet to which raw or heated CNSL was added at a final concentration of 500 µg/mL. In experiment 2, a Rusitec, using rumen fluid from the same cattle, was operated over a period of 7 d during which only raw CNSL was tested at concentrations of 0, 50, 100, or 200 µg/mL, and variations in fermentation and bacterial population were assessed. In experiment 3, a pure culture study was conducted using selected bacteria to determine their susceptibility to CNSL. In experiment 1, methane production was inhibited by raw CNSL (56.9% inhibition) but not by heated CNSL. Total volatile fatty acid concentration was not affected, whereas increased concentrations of propionate and decreased concentrations of acetate and butyrate were observed using either raw or heated CNSL. These changes were more obvious when raw CNSL was tested. In experiment 2, raw CNSL inhibited methanogenesis and increased propionate production in a dose-dependent manner, showing maximum methane inhibition (70.1%) and propionate enhancement (44.4%) at 200 µg/mL supplementation. Raw CNSL increased total volatile fatty acid concentration and dry matter digestibility. Raw CNSL also appeared to induce a dramatic shift in the population of rumen microbiota, based on decreased protozoa numbers and changes in quantitative PCR assay values for representative bacterial species. In experiment 3, using pure cultures, raw CNSL prevented the growth of hydrogen-, formate-, and butyrate-producing rumen bacteria, but not the growth of bacteria involved in propionate production. Based on these data, raw CNSL, rich in the antibacterial phenolic compound anacardic acid, is a potential candidate feed additive with selective activity against rumen microbes, leading to fermentation that results in decreased methane and enhanced propionate production.

Vitamin B6 deficiency and anemia in pregnancy.

Iron deficiency is the most common cause of anemia in pregnancy. Pregnant women with anemia are, in general, exclusively treated with iron supplementation. We observed that several pregnant women with anemia who were nonresponsive to iron supplementation also had vitamin B6 deficiency, and that anemia in these cases improved with the administration of vitamin B6. Our prospective study in healthy pregnant women showed that blood levels of iron, ferritin and vitamin B6, in particular, fell to the lower limit of the nonpregnant reference range by the third trimester. We conclude that it is important to take into account the deficiency of vitamin B6 besides iron in the evaluation of anemia during pregnancy.

Dietary effects of bitter gourd oil on blood and liver lipids of rats.

Bitter gourd is widely used as an edible plant in Asia. In this study, we evaluated the effects of bitter gourd oil (BGO) on the blood and liver lipids of rats. Three groups of rats were given a basal diet (AIN-93G) containing 7% fat by weight. The dietary fat consisted of soybean oil (control), soybean oil + BGO (6.5:0.5, w/w; 0.5% BGO), or soybean oil + BGO (5:2, w/w; 2.0% BGO). This fat treatment gave 3.4 and 15.4% of cis(c)9,trans(t)11,t13-18:3 in the dietary fat of 0.5 and 2.0% BGO, respectively. Fatty acid analysis showed the occurrence of c9,t11-18:2 in the liver of rats fed BGO diets, whereas this conjugated linoleic acid (CLA) isomer was not detected in the liver of rats fed the control diet. Furthermore, dietary BGO decreased the concentration of 18:2n-6 and increased the concentration of 22:6n-3. The formation of the CLA isomer in the liver lipids of rats fed BGO diets could be explained by either of the following two metabolic pathways, namely, enzymatic biohydrogenation of c9,t11,t13-18:3 or enzymatic isomerization of c9,c12-18:2. The BGO diets had significantly reduced free cholesterol levels with a trend toward an increase in HDL cholesterol, but there was no significant change in the total cholesterol. The dietary BGO also affected the level of plasma hydroperoxides. A slight but significant increase in hydroperoxides was found in the rats fed 2.0% BGO. This may be attributed to the lower oxidative stability of c9,t11,t13-18:3 in BGO.

Cytotoxic effect of conjugated trienoic fatty acids on mouse tumor and human monocytic leukemia cells.

The cytotoxicity of fatty acids from seed oils containing conjugated linolenic acids (CLN) was studied. Fatty acids from pomegranate, tung, and catalpa were cytotoxic to human monocytic leukemia cells at concentrations exceeding 5 microM for pomegranate and tung and 10 microM for catalpa, but fatty acids from pot marigold oil had no effect at concentrations ranging up to 163 microM. The main conjugated fatty acids of pomegranate, tung, catalpa, and pot marigold were cis(c)9,trans(t)11,c13-CLN (71.7%), c9,t11,t13-CLN (70.1%), t9,t11,c13-CLN (31.3%), and t8,t10,c12-CLN (33.4%), respectively. Therefore, the cytotoxicities of fatty acids from pomegranate, tung, and catalpa were supposed to be due to 9,11,13-CLN isomers. To elucidate the cytotoxicity of these CLN, we separated each CLN isomer from the fatty acid mixtures by high-performance liquid chromatography and analyzed its cytotoxicity. The cytotoxicities of c9,t11,c13-CLN, c9,t11,t13-CLN, and t9,t11,c13-CLN were much stronger than that of t8,t10,c12-CLN. Therefore, the higher cytotoxicity of fatty acids from pomegranate, tung, and catalpa than those from pot marigold would be derived from the different activities of 9,11,13-CLN and 8,10,12-CLN. Since there was little difference in the cytotoxicities of c9,t11,c13-CLN,c9,t11,t13-CLN, and t9,t11,c13-CLN, it is suggested that the cis/trans configuration of 9,11,13-CLN isomers had little effect on their cytotoxic effects. The mechanism of the cytotoxicity of the four fatty acids above may involve lipid peroxidation, because the order of toxicity of the fatty acids was consistent with their susceptibility to peroxidation in aqueous phase. This was supported by the decrease in the cytotoxicity of the fatty acids by addition of butylated hydroxytoluene.

Activating mutation of D835 within the activation loop of FLT3 in human hematologic malignancies.

Mutations of receptor tyrosine kinases are implicated in the constitutive activation and development of human malignancy. An internal tandem duplication (ITD) of the juxtamembrane (JM) domain-coding sequence of the FLT3 gene (FLT3/ITD) is found in 20% of patients with acute myeloid leukemia (AML) and is strongly associated with leukocytosis and a poor prognosis. On the other hand, mutations of the c-KIT gene, which have been found in mast cell leukemia and AML, are clustered in 2 distinct regions, the JM domain and D816 within the activation loop. This study was designed to analyze the mutation of D835 of FLT3, which corresponds to D816 of c-KIT, in a large series of human hematologic malignancies. Several kinds of missense mutations were found in 30 of the 429 (7.0%) AML cases, 1 of the 29 (3.4%) myelodysplastic syndrome (MDS) cases, and 1 of the 36 (2.8%) acute lymphocytic leukemia patients. The D835Y mutation was most frequently found (22 of the 32 D835 mutations), followed by the D835V (5), and D835H (1), D835E (1), and D835N (1) mutations. Of note is that D835 mutations occurred independently of FLT3/ITD. An analysis in the 201 patients newly diagnosed with AML (excluding M3) revealed that, in contrast to the FLT3/ITD mutation (n = 46), D835 mutations (n = 8) were not significantly related to the leukocytosis, but tended to worsen disease-free survival. All D835-mutant FLT3 were constitutively tyrosine-phosphorylated and transformed 32D cells, suggesting these mutations were constitutively active. These results demonstrate that the FLT3 gene is the target most frequently mutated to become constitutively active in AML.

[Three long surviving patients with gastric cancer metastasizing to the liver under interdisciplinary therapy].

In 30 patients with gastric cancer metastasizing to the liver over the past 15 years at our hospital the primary foci in the stomach could be resected in a curative manner. The authors report herein three long surviving patients in this series. [Case 1] A 49-year-old male. Distal gastrectomy was performed in November 1984. Metastasis to the liver occurred in June 1986. The right lobe of the liver was resected in November 1987 after transcatheter arterial embolization (TAE). Although hepatic arterial infusion chemotherapy was conducted, the cancer metastasized to the whole body, and the patient died in December 1991. [Case 2] A 65-year-old female. Distal gastrectomy was performed in July 1994. The left hepatic lobe and segment 5 in the right lobe were resected in June 1995. Although TAE was performed six times starting in December 1996, the patient died of hepatic failure in July 1999. [Case 3] A 73-year-old male. This patient simultaneously received distal gastrectomy and extended resection of the posterior hepatic segments in September 1997. Cancer recurred in the remaining liver in July 1998. Although microwave coagulation therapy (MCT) and TAE were performed, the patient died of hepatic failure in January 2000. In these patients who survived for a long period, the primary focus was well-differentiated adenocarcinoma under sufficient local control with metastasis limited to the nearest regional lymph nodes (group 1 lymph nodes). The patients could undergo interdisciplinary therapy, including hepatectomy, MCT, TAE, and hepatic arterial infusion chemotherapy.

High throughput screening for human interferon-gamma production inhibitor using homogenous time-resolved fluorescence.

An immunoassay for interferon-gamma (IFN-gamma) using homogeneous time-resolved fluorescence (HTRF) has been developed. In this assay, IFN-gamma can be detected by simply adding a mixture of three reagents-biotinylated polyclonal antibody, europium cryptate (fluorescence donor, EuK)-labeled monoclonal antibody, and crosslinked allophycocyanin (fluorescence acceptor, XL665) conjugated with streptavidin-and then measuring the time-resolved fluorescence. The detection limit of IFN-gamma by the proposed method is about 625 pg/ml. We applied the method to the detection of IFN-gamma secreted from NK3.3 cells and employed it in high throughput screening for IFN-gamma production inhibitors. With this screening format, IFN-gamma can be measured by directly adding the above reagents to microplate wells where NK3.3 cells are being cultured and stimulated with interleukin-12. This "in situ" immunoassay requires only pipetting reagents, with no need to transfer the culture supernatant to another microplate or wash the plate. Therefore, this screening format makes possible full automation of cell-based immunoassay, thus reducing cost and experimental time while increasing accuracy and throughput.

Characterization of native and recombinant peptidyl prolyl cis-trans isomerases derived from Methanococcus thermolithotrophicus based on cDNA sequence.

It is important to establish whether a recombinant protein is an authentic copy of the predicted cDNA sequence. In this study, recombinant protein for native peptidyl prolyl cis-trans isomerase (N-PPIase) and double-labeled (13C- and 15N-) protein (DL-PPIase) appeared on the sodium dodecyl sulfate (SDS) electropherograms as two bands for N-PPIase and four bands for DL-PPIase. Since the N-terminal amino acid residues of all bands were the same, we characterized these bands using the peptide mapping method and amino acid composition analysis. Peptide mapping of the proteins seemed to be almost identical but they could not reflect the whole amino acid sequences of the protein. The bands on the polyvinylidene difluoride (PVDF) membrane, electroblotted after SDS-polyacrylamide gel electrophoresis (SDS-PAGE), were hydrolyzed and their amino acid composition was analyzed using a highly sensitive 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) amino acid analysis and compared with the cDNA sequences for proteins. The matching score (sigma(T%-E%)2) for similarity of proteins was calculated by summation of the square difference between the theoretical (T%) and the experimental (E%) amino acid composition of the recombinant protein. The amino acid composition of all bands of both proteins showed more than 93% of the theoretical values. The major molecular weights of both proteins were 16812 and 17694 by electrospray ionization (ESI)-mass spectrometry. However, the purified proteins also contained minor compounds with Mr of 3721 for N-PPIase and 5285 for DL-PPIase. These compounds were considered to be nonpeptidyl products that comigrated with the protein. Similarities of the amino acid composition of the four bands were more than 98%. Our results indicate that AQC amino acid analysis is the most suitable method for characterization of a recombinant protein.

Novel antiarthritic agents with 1,2-isothiazolidine-1,1-dioxide (gamma-sultam) skeleton: cytokine suppressive dual inhibitors of cyclooxygenase-2 and 5-lipoxygenase.

Various 1,2-isothiazolidine-1,1-dioxide (gamma-sultam) derivatives containing an antioxidant moiety, 2,6-di-tert-butylphenol substituent, were prepared. Some compounds, which have a lower alkyl group at the 2-position of the gamma-sultam skeleton, showed potent inhibitory effects on both cyclooxygenase (COX)-2 and 5-lipoxygenase (5-LO), as well as production of interleukin (IL)-1 in in vitro assays. They also proved to be effective in several animal arthritic models without any ulcerogenic activities. Among these compounds, (E)-(5)-(3,5-di-tert-butyl-4-hydroxybenzylidene)-2-ethyl-1, 2-isothiazolidine-1,1-dioxide (S-2474) was selected as an antiarthritic drug candidate and is now under clinical trials. The structure-activity relationships (SAR) examined and some pharmacological evaluations are described.

Modulation of lung local immune responses by oral administration of a herbal medicine Sho-saiko-to.

Sho-saiko-to (SST), a Chinese/Japanese herbal medicine (Kampo medicine) widely used to treat chronic hepatitis in Japan, is known to modulate immune responses, and thus its immunomodulating activity may be responsible for its bi-directional effects on the lungs as therapeutic efficacy in various lung diseases and involvement in development of interstitial pneumonia. We administered SST to BALB/c mice orally and examined the lung tissue levels of pro/anti-inflammatory cytokines, interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6), and the effects of SST on acute lung injury induced by instillation of lipopolysaccharide (LPS) or IL-1. Although SST had no effect on lung TNF-alpha or IL-1beta level, it increased IL-6. Investigation of active fractions of SST suggested that multiple ingredients were supposed to be responsible for IL-6-inducing activity. Liquiritigenin, a metabolite of liquiritin which is one of the major ingredients in SST enhanced in vitro IL-6 production in anti-CD3 monoclonal antibody (anti-CD3 mAb)-stimulated lung mononuclear cells in a cell-type specific and dose-dependent manner. SST suppressed LPS-induced lung injury at the later phase when lung leak was evident while being ineffective on initial neutrophil sequestration to the lung in these models. These findings suggest that SST modulates lung inflammation by regulating local immune response.

Synthesis of a biologically active fluorescent derivative of GM1, a main Ginseng saponin metabolite formed by intestinal bacteria.

A fluorescent derivative of GM1 [20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol], a main Ginseng saponin metabolite formed by intestinal bacteria, was obtained from the condensation of its trisnor-aldehyde derivative with dansyl hydrazine. The dansylated GM1 fluoresced strongly and showed almost the same properties as its parent compound in lipophilicity and biological activities, so this fluorescent compound might provide an insight into the mechanism of pharmacological activities of GM1.

Cell adhesion kinase beta forms a complex with a new member, Hic-5, of proteins localized at focal adhesions.

Cell adhesion kinase beta (CAKbeta/PYK2) is the second protein-tyrosine kinase of the focal adhesion kinase subfamily. We identified a cDNA that encodes a CAKbeta-binding protein. This cDNA clone encodes the human homologue of Hic-5, the cDNA of which was cloned in 1994 as transforming growth factor beta1- and hydrogen peroxide-inducible mRNA. We found that Hic-5 exclusively localized at focal adhesions in a rat fibroblast line, WFB. This localization of Hic-5 was confirmed in WFB cells expressing Myc-tagged Hic-5. The amino acid sequence of Hic-5 is highly similar to that of paxillin in the four LD motifs as well as in the four contiguous LIM domains. The Hic-5 N-terminal domain directly associated in vitro with the extreme C-terminal region (residue 801 to the end) of CAKbeta. CAKbeta was coimmunoprecipitated with Hic-5 from the WFB cell lysate. The coimmunoprecipitation of CAKbeta with Hic-5 was markedly inhibited by the addition of the extreme C-terminal region of CAKbeta. Coimmunoprecipitation of Hic-5 with CAKbeta, which was shown in COS-7 cells doubly transfected with cDNA constructs of CAKbeta and Myc-tagged Hic-5, was lost when the CAKbeta amino acid residues 741-903 were deleted. Hic-5 was tyrosine-phosphorylated in Src-transformed 3Y1 cells and in cells treated with pervanadate. Hic-5 associated with CAKbeta was selectively tyrosine-phosphorylated in WFB cells exposed to hypertonic osmotic stress. These results indicate that Hic-5 is a paxillin-related component of focal adhesions and binds to CAKbeta, implying possible involvement of Hic-5 in the downstream signaling of CAKbeta.

Vesnarinone prolongs action potential duration without reverse frequency dependence in rabbit ventricular muscle by blocking the delayed rectifier K+ current.

BACKGROUND: Methanesulfonanilide derivatives, selective inhibitors of the rapidly activating component (I(Kr)) of the delayed rectifier potassium current (I(K)), prolong action potential duration (APD) of cardiac muscles with reverse frequency dependence, which limits their clinical use because of proarrhythmia. Vesnarinone, a quinolinone derivative developed as a cardiotonic agent, has complex pharmacological properties, but its clinical efficacy is explained in part by I(K) reduction. Therefore, we investigated the mode of I(K) block by vesnarinone. METHODS AND RESULTS: I(K) of the rabbit ventricular myocyte was activated by voltage-clamp steps applied from a holding potential to various depolarizing levels. The development of I(K) block at depolarization (+10 mV) and its recovery process at hyperpolarization (-75 mV) were compared between vesnarinone and E-4031. The I(K) block by vesnarinone (3 micromol/L) developed and recovered monoexponentially, with time constants of 361 ms (n=5) and 1.87 seconds (n=4), respectively. I(K) block by E-4031 (0.3 micromol/L) developed instantaneously, with no recovery from the block at hyperpolarization. The I(K) block by vesnarinone, estimated by I(K) tail after a train of depolarizing pulses (for 30 seconds at 0.2 to 2 Hz), was increased with increasing frequency (twofold at 2 from 0.2 Hz), but that by E-4031 was unchanged. In rabbit papillary muscles, vesnarinone (10 micromol/L) prolonged APD at stimulation frequencies >0.2 Hz, whereas E-4031 (0.3 micromol/L) prolonged that in a reverse frequency-dependent manner. CONCLUSIONS: Vesnarinone may prolong the repolarization of human cardiac muscle without reverse frequency dependence, because I(Kr) is expressed in humans as well as in the rabbit. Thus, this drug may be a model for an ideal class III drug without the risk of proarrhythmia.

Synthesis and evaluation of novel fluorinated methotrexate derivatives for application to rheumatoid arthritis treatment.

An ongoing search for new antifolate drugs useful against rheumatoid arthritis (RA) led us to prepare new methotrexate (MTX) derivatives containing enantiomerically pure L-erythro- or L-threo-gamma-fluoroglutamic acid. The derivatives in which the phenyl ring was replaced by a 3'-substituted phenyl or methylthiophene ring showed potent immunosuppressive activities, including in vitro inhibition of mitogen responses to both T and B cells and in vivo inhibition of antibody production in mice. These compounds also exhibited inhibitory activity in adjuvant arthritis in rats. Their toxicity was lower than that of MTX, which was probably due to the strong electronegativity of fluorine, which increases the acidity of the gamma-carboxyl group and thereby decreases polyglutamylation in normal cells. These results revealed the potential of the fluorinated MTX derivatives as candidate drugs for the treatment of RA.

Meningoencephalocele associated with Tripterygium wilfordii treatment.

We treated a male infant with occipital meningoencephalocele associated with the taking of Tripterygium wilfordii. The infant was delivered normally at 38 weeks of gestation with a huge cystic mass protruding from the occiput. He was diagnosed with occipital meningoencephalocele and cerebellar agenesis. His mother had taken T. wilfordii for rheumatoid arthritis early in her pregnancy. T. wilfordii is a herbal medicine used for rheumatoid arthritis and male contraception. Since its toxicity is high and its use during pregnancy is restricted, it is the most likely cause of this infant's anomalies.

A comparison of blood pressure changes in phacoemulsification cataract surgery with topical and retrobulbar block local anesthesia.

BACKGROUND: Changes are observed in blood pressure (BP) levels during cataract surgery, although BPs are considered to remain stable under local anesthesia. We evaluated the daily, pre- and postoperative BPs of 2270 patients after cataract surgery performed under either topical anesthesia or retrobulbar block. METHODS: All operations were performed by the same surgeon using the same method of phacoemulsification and aspiration with posterior chamber intraocular lens implantation under local anesthesia. RESULTS: The mean daily BP was 99.3 +/- 14.2 mm Hg; the mean preoperative BPs increased and then the postoperative BPs decreased. The postoperative BPs of the retrobulbar injection group decreased significantly more than those of the topical application group. In 833 cases, the systolic BP changed by more than 20 mm Hg. Even when the patients were hypertensive, the preoperative and postoperative BPs decreased in the same manner. CONCLUSION: The present study shows that, following surgery with retrobulbar block anesthesia, BP decreases to a greater extent than with topical anesthesia. Physicians should be aware of the high proportion of cases in which the systolic BP changes by more than 20 mm Hg.

Inhibitory effects of pituitary adenylate cyclase activating polypeptide on histamine-induced respiratory resistance in anesthetized guinea pigs.

Asthma is a chronic inflammatory disorder of the airways in which many cells participate. This inflammation causes recurrent episodes and symptoms that are associated with widespread but variable airflow limitation that is at least partly reversible either spontaneously or with treatment. Therefore, an investigation of useful remedies for the treatment of bronchial asthma is proposed. In this study, we determined whether both forms of pituitary adenylate cyclase activating polypeptide (PACAP 38 and PACAP 27) belonging to the vasoactive intestinal peptide (VIP) family of peptides could inhibit the effects of histamine-induced respiratory resistance (Rr) in anesthetized guinea pigs, when compared with VIP. The order for 50% suppression (ED50) of Rr induced by peptides was VIP > PACAP 27 > PACAP 38. The inhibitory effects induced by PACAP 38 on histamine-induced Rr in guinea pigs were more prolonged than with the other two peptides. Moreover, adding the endopeptidase inhibitor phosphoramidon prolonged the inhibitory effects of PACAPs. These results suggested that the exogenous peptides of the inhibitory nonadrenergic noncholinergic nervous (i-NANC) peptides could become a useful remedy for treatment of bronchial asthma, because these belong to an important intrinsic hormone.

[Studies on enrichment broth for verotoxin-producing Escherichia coli (VTEC) O157].

Growth of verotoxin-producing Escherichia coli (VTEC) O157 in conventionally recommended pre-enrichment broth media at different temperatures was evaluated. In addition, sensitivity of VTEC O157 isolates to several antibacterial drugs, which were added to the selective enrichment broth, was tested. All five isolates of VTEC O157 tested grew well in trypticase soy broth (TSB) at 36 degrees C and 42 degrees C, while the growth of one isolate was markedly suppressed in TSB supplemented with cefixime (CFIX), potassium tellurite (PT), and vancomycin (TSB-CTV) even at 36 degrees C. A significant growth suppression was also observed in three of the isolates cultured in novobiocin (NB)-supplemented modified EC broth (mEC-NB) at 42 degrees C. In mEC-NB after 24-hr incubation at 36 degrees C, the five VTEC O157 isolates grew well, although one isolate was slightly suppressed during the first 8 hours. Minimum growth inhibitory concentrations of CFIX, NB and PT for a total of 90 clinical and environmental isolates of VTEC O157 were all above the concentrations usually prescribed for mEC-NB and TSB-CTV. These findings suggest that mEC-NB and TSB-CTV should be used at 36 degrees C for growth of VTEC O157 and that use of a nonselective pre-enrichment broth medium (i.e. TSB) together with a selective one (i.e. TSB-CTV or mEC-NB) is necessary for successful isolation of VTEC O157 from various specimens.