SEPT5 Research Articles

Antibacterial activity of jackfruit leaves extracts and the interference on antimicrobial susceptibility of enteropathogen

The objective of the present study was to evaluate the antimicrobial potential of Artocarpus heterophyllus dry leaves extracts on S. enterica and E. coli and their interaction with conventional antimicrobials. Dried powdered leaves were used to produce hexane (Hex), methanolic (MeOH) and ethanolic residue (EtOHr) extracts. The antimicrobial test was performed against Escherichia coli, ATCC 25922, E. coli EPEC, CDC 086H35, and Salmonella enterica serotype Enteritidis phagotype 4 (SE PT4) through minimal inhibition concentration (MIC) and minimal bactericidal concentration (MBC). The agar diffusion technique, well and disc-variants were used to measure the antimicrobial effect of the plant extract combination with antimicrobial of clinical usage. We highlight the bactericidal effect of jackfruit tree leaves on E. coli and S. enterica SE PT4 at 7.2 mg/mL and the effect of the extracts on antimicrobial activity. The interaction between chloramphenicol and dry leaves extracts was characterized by both synergism and antagonism depending on extract type and bacteria used. The interaction between antimicrobial of clinical usage and jackfruit tree leaves extracts demonstrated changes in susceptibility profile of antimicrobials tending to an antagonist effect. As the jackfruit tree leaves may interfere on antimicrobials action, special attention should be given to its usage as traditional medicine in the treatment of food borne diseases.

MiR-185 and SEPT5 Genes May Contribute to Parkinson's Disease Pathophysiology.

There are still unknown mechanisms involved in the development of Parkinson's disease (PD), which elucidating them can assist in developing efficient therapies. Recently, studies showed that genes located on the human chromosomal location 22q11.2 might be involved in the development of PD. Therefore, the present study was designed to evaluate the role of two genes located on the chromosomal location (miR-185 and SEPT5), which were the most probable candidates based on our bibliography. In vivo and in vitro models of PD were developed using male Wistar rats and SHSY-5Y cell line, respectively. The expression levels of miR-185, SEPT5, LRRK2, and PARK2 genes were measured at a mRNA level in dopaminergic areas of rats' brains and SHSY-5Y cells using the SYBR Green Real-Time PCR Method. Additionally, the effect of inhibition on the genes or their products on cell viability and gene expression pattern in SHSY-5Y cells was investigated. The level of miR-185 gene expression was significantly decreased in the substantia nigra (SN) and striatum (ST) of the rotenone-treated group (control group) compared to the healthy normal group (P < 0.05). In addition, there was a significant difference in the expression of SEPT5 gene (P < 0.05) in the substantia nigra between two studied groups. The results of an in vitro study showed no significant change in the expression of the genes; however, the inhibition on miR-185 gene expression led to the increase in LRRK2 gene expression in SHSY-5Y cells. The inhibition on LRRK2 protein also decreased the cellular toxicity effect of rotenone on SHSY-5Y cells. The results suggested the protective role of miR-185 gene in preventing the development of PD.

Interaction of Recombinant Gallus gallus SEPT5 and Brain Proteins of H5N1-Avian Influenza Virus-Infected Chickens.

Septin forms a conserved family of cytoskeletal guanosine triphosphate (GTP) binding proteins that have diverse roles in protein scaffolding, vesicle trafficking, and cytokinesis. The involvement of septins in infectious viral disease pathogenesis has been demonstrated by the upregulation of SEPT5 protein and its mRNA in brain tissues of H5N1-infected chickens, thus, providing evidence for the potential importance of this protein in the pathogenesis of neurovirulence caused by the avian influenza virus. In this study, cloning, expression, and purification of Gallus gallus SEPT5 protein was performed in Escherichia coli. The SEPT5 gene was inserted into the pRSETB expression vector, transformed in the E. coli BL21 (DE3) strain and the expression of SEPT5 protein was induced by IPTG. The SEPT5 protein was shown to be authentic as it was able to be pulled down by a commercial anti-SEPT5 antibody in a co-immunoprecipitation assay. In vivo aggregation of the recombinant protein was limited by cultivation at a reduced temperature of 16 °C. Using co-immunoprecipitation techniques, the purified recombinant SEPT5 protein was used to pull down host’s interacting or binding proteins, i.e., proteins of brains of chickens infected with the H5N1 influenza virus. Interacting proteins, such as CRMP2, tubulin proteins, heat-shock proteins and other classes of septins were identified using LCMS/MS. Results from this study suggest that the codon-optimized SEPT5 gene can be efficiently expressed in the E. coli bacterial system producing authentic SEPT5 protein, thus, enabling multiple host’s proteins to interact with the SEPT5 protein.

Interventions to reduce the bacterial load in recycled broiler litter

Two experiments were undertaken to evaluate the bacterial load in recycled litter between broiler flocks following addition of quicklime (T1), windrowing (T2), shallow fermentation (T3), and control (no intervention, T4). The first experiment was developed in field conditions in which the broiler houses were accompanied by 6 consecutive flocks and the effect of the treatments was assessed on enterobacteria and aerobic mesophiles. The second experiment was conducted in an experimental broiler house with recycled litter for assessment of Salmonella Enteritidis phage type 4 (SE PT4). In the field study, T3 presented the best results in reducing enterobacteria in broiler litter in relation to the other treatments, with the highest reduction occurring in the first 3 flocks, tending to stabilization from the fourth flock onward for all the treatments assessed. From the third to sixth flocks, enterobacteria level at the end of the treatments (d 12) was lower than the average in the fresh litter, except in T4. All treatments reduced aerobic mesophiles throughout the flocks, where T2 showed the highest reduction. The percentage of dry matter in the broiler litter diminished in T4 and increased in T3 over the course of the flocks. In the second experiment, the drop in the SE PT4 level in the broiler litter first occurred in T2 and T3. However, all the treatments except for T4 eliminated SE PT4 within 12 d. The temperature of the broiler litter in T2 was higher in relation to the other treatments. The results show that litter treatment prior to reutilization by the successive broiler flock is required to reduce the level of residual bacteria. The fermentative treatments (T2 and T3) were found to be superior to the others in terms of reducing the bacterial load, with shallow fermentation standing out with the highest reduction of enterobacteria and equivalent SE PT4 elimination when compared to windrowing.

Effect of Enterococcus faecium EF 55 on morphometry and proliferative activity of intestinal mucosa in broilers infected with Salmonella Enteritidis

Abstract Introduction: The present study aimed to investigate the effect of Enterococcus faecium EF55 on chickens, as well as its influence on proliferative activity of epithelial intestinal cells after infection with Salmonella enterica serovar Enteritidis phage type 4 (SE PT4). Moreover, the length and area of duodenal and jejunal villi of the birds were examined. Material and Methods: A pool of 80 birds was divided randomly into four groups. Probiotic group (EF) and Salmonella + probiotic group (EFSE) received E. faecium EF55 (109 CFU – 3 g per group/day) during 22 d. Salmonella group (SE) and EFSE group were infected with Salmonella Enteritidis PT4 (108 CFU in 0.2 mL PBS) in a single dose per os on day four of the experiment, whereas control birds (C group) received only 0.2 mL of PBS on that day. Samples were taken on the 4th and 18th day post infection. Results: Supplementation of feed with E. faecium EF 55 confirmed its selective antibacterial activity against SE PT4. The chickens infected with SE PT4 and fed E. faecium EF55 supplemented diet showed increased proliferative activity of enterocytes in the jejunum in both samplings. Applied probiotic strain demonstrated positive impact on intestinal morphometry in the jejunum of both non-infected groups and in Salmonella-infected chickens. In the latter group, the beneficial effect of E. faecium EF 55 was manifested by more efficient tissue turnover in the jejunum.

MLL-SEPT5 fusion transcript in infant acute myeloid leukemia with t(11;22)(q23;q11)

Chromosomal rearrangements involving the MLL gene at band 11q23 are the most common genetic alteration encountered in infant acute myeloid leukemia. Reciprocal translocation represents the most frequent form of MLL rearrangement. Currently, more than 60 partner genes have been identified. We report here a case of de novo acute myeloid leukemia with a t(11;22)(q23;q11) in a 23-month-old child. Fluorescence in situ hybridization study revealed that the 3’MLL segment was translocated onto the derivative chromosome 22 and the breakpoint on chromosome 22 was located in or near the SEPT5 gene at 22q11.21. Long distance inverse-polymerase chain reaction was used to identify precisely the MLL partner gene and confirmed the MLL–SEPT5 fusion transcript. Involvement of the SEPT5 gene in MLL rearrangement occurs very rarely. Clinical, cytogenetic and molecular features of acute myeloid leukemia with a MLL–SEPT5 fusion gene are reviewed.

Septin 4 (SEPT4; ARTS)

Septin 4 (SEPT4; ARTS)

The Alteration of SEPT5 Gene Expression in BCR-ABL Positive Cells and Its Possible Correlation with the Development and / or Progression of Chronic Myeloid Leukemia (CML)

Abstract 4415The CML is a clonal disease of stem cells and its main feature is the unregulated production of a tyrosine kinase protein called BCR-ABL, the progression of the disease to accelerated phase or blast crisis may be associated with genomic instability. Because of this, the use of tools for the study of gene expression could bring new insights in the understanding of these mechanisms in the CML. In a recent study using SSH libraries, we compared the gene expression pattern between granulocytes of health control and CML patients, and we identified the gene SEPT5 expressed only in CML patients. Although the studies in the literature, there is not a clear relationship between the expression of this gene and the development or progression of CML. SEPT5 is a member of nucleotide binding proteins called septins that were firstly described in yeast as cell division cycle regulatory proteins. This gene was reported in patients with AML translocated with MLL gene, in adult human brain and heart; it is also associated with alpha granules of human blood platelets. The aims of this study are to carry a functional analysis of SEPT5 in differents cells line and to study the relationship of this gene and the development and/or progression of CML. The gene expression evaluation was made in granulocytes, mononuclear cells and total leukocytes of CML patients and healthy blood donors in peripheral blood. It was also evaluated in bone marrow donors, in human cell lines (K562, HL60 and NB4) and in mice cell lines (BaF3/BCR-ABLp210 and BaF3T315I), performed by real-time PCR for the following genes: SEPT5, β-actin and GAPDH. Experiments were also performed to verify the difference between the chemotaxis of granulocytic cells from controls and patients by ELISA. Data were analysed statistically using the ANOVA followed by Dunnett's test – P value of less than 0.05 was considered to be significant. The study was approved by the Research Ethic Committee of the Faculty of Medical Sciences of University of Campinas. The gene expression of SEPT5 was evaluated by real time PCR using the same samples used in the library construction to validate the results found in the SSH library. The data confirmed our previous results, showing that the SEPT5 expression is increased in all cells of patients compared to controls. The same results were observed when we studied the expression comparing individually patients and health blood donors, suggesting that this protein could be increased in all human cells that present the translocation BCR-ABL. The level of expression of this gene in HL60 and NB4 was significantly lower than in K562 cell line. The experiments with mice cell lines showed a higher expression of this gene in BaF3T315I when compared to BaF3BCR-ABLp210. We obtained a significant expression difference in all experiments (p <0.05). The spontaneous and stimulated with IL-8 chemotaxis assays used granulocytes and were assessed using chamber containing 96 wells. However, although the results suggest an increased chemotactic activity in patients, there were no significant differences (p<0.05) between controls and patients – regardless of whether the chemotaxis was spontaneous or stimulated with IL-8. In mammals the SEPT5 gene is associated with cellular processes such as exocytosis, apoptosis, leukemogenesis, carcinogenesis and neurodegeneration. Therefore, molecules capable of interacting with the septins, either at biochemical or molecular level, can bring information about their functions in cytokinesis. Studies indicate that the human septins can interact among themselves and with other components of the cytoskeleton – this may be a relevant observation regarding the function of this gene in cancer. The SEPT5 can be activated by different pathways – this may increase expression in translocated cells. Despite major advances in the treatment of CML, the treatments available are not capable of inactivating all the signaling pathways activated by BCR/ABL. Our results demonstrate that SEPT5 may be involved in the pathophysiology of CML. Also, it is clear the importance of the study of pathways that could culminate in its high expression or the triggering of other unknown pathways involved in the development of CML. The increased expression of this gene may be related to disease progression, and finally, the identification of several important genes may lead to a better understanding of CML and helping to identify new therapeutic targets. FAPESP/INCT. Disclosures:No relevant conflicts of interest to declare.

Leukocytic responses and intestinal mucin dynamics of broilers protected with Enterococcus faecium EF55 and challenged with Salmonella Enteritidis

The protective effect of Enterococcus faecium EF55 in chickens challenged with Salmonella enterica serovar Enteritidis phage type 4 (SE PT4) was assessed. The antibacterial effect on the bacterial microflora in the small intestine in relation to white blood cell count, phenotyping of peripheral blood and intestinal lymphocytes, functional activity of lymphocytes and phagocytes and mucin quantitation were investigated. Day-old chicks (85) were randomly divided into four groups. The probiotic group (EF) and Salmonella+probiotic group (EFSE) received E. faecium EF55 (109 CFU – 3g/group/day) for 21days. The Salmonella group (SE) and EFSE group were infected with Salmonella Enteritidis (108 CFU in 0.2ml PBS) in a single dose per os on day four of the experiment. The control group chicks (C) were fed a commercial diet without added bacteria. Supplementation of EF55 in the diet of the chickens in the EFSE group, challenged with S. Enteritidis, caused the density of the intestinal mucin layer to increase significantly in non-specific regions (duodenum and jejunum), but decrease significantly in target regions (caeca) for S. Enteritidis. Probiotic treatment also appeared to result in a significantly higher number of lymphocytes in peripheral blood and a tendency to increase CD3, CD4, CD8, and IgM positive cells 3days post-infection with S. Enteritidis. The results demonstrated an antibacterial effect and suggested that EF55 had a moderating effect on intestinal mucin production and leukocytic response in the early phase of S. Enteritidis infection.

Deletion of human GP1BB and SEPT5 is associated with Bernard-Soulier syndrome, platelet secretion defect, polymicrogyria, and developmental delay

The bleeding disorder Bernard-Soulier syndrome (BSS) is caused by mutations in the genes coding for the platelet glycoprotein GPIb/IX receptor. The septin SEPT5 is important for active membrane movement such as vesicle trafficking and exocytosis in non-dividing cells (i.e. platelets, neurons). We report on a four-year-old boy with a homozygous deletion comprising not only glycoprotein Ibβ (GP1BB) but also the SEPT5 gene, located 5' to GP1BB. He presented with BSS, cortical dysplasia (polymicrogyria), developmental delay, and platelet secretion defect. The homozygous deletion of GP1BB and SEPT5, which had been identified by PCR analyses, was confirmed by Southern analyses and denaturing HPLC (DHPLC). The parents were heterozygous for this deletion. Absence of GPIbβ and SEPT5 proteins in the patient's platelets was illustrated using transmission electron microscopy. Besides decreased GPIb/IX expression, flow cytometry analyses revealed impaired platelet granule secretion. Because the bleeding disorder was extremely severe, the boy received bone marrow transplantation (BMT) from a HLA-identical unrelated donor. After successful engraftment of BMT, he had no more bleeding episodes. Interestingly, also his mental development improved strikingly after BMT. This report describes for the first time a patient with SEPT5 deficiency presenting with cortical dysplasia (polymicrogyria), developmental delay, and platelet secretion defect.

Antimicrobial activity of Enterococcus faecium EF 55 against Salmonella Enteritidis in chicks

The protective effect of Enterococcus faecium EF 55 against Salmonella enterica serovar Enteritidis phage type 4 (SE PT4) was studied in 1-day-old chicks. The EF 55 strain (isolated and characterised by the authors earlier) was applied daily (1.10(9) CFU/0.2 ml PBS) for 7 days. Oral inoculation of the SE PT4 strain was performed on day 8 in a single dose of 5.10(8) CFU/0.2 ml PBS. The experiment lasted for 21 days. Samples were collected on day 1 of the experiment to verify the absence of Salmonella, on day 8 to check colonisation of EF 55 and immunological status in experimental birds, and on days 2, 4, 6, 8 and 14 after SE PT4 infection of chicks. Strain EF 55 sufficiently colonised the digestive tract of chicks after 7 days of application. The highest numbers of EF 55 in the faeces of chicks were observed before SE infection and persisted to day 6 post infection (p.i.) in both the EF and EF+SE groups. PCR confirmed the identity of the EF 55 strain. The counts of SE PT4 strain in faeces of the EF+SE group were significantly reduced in comparison to those in the SE group on days 2 and 14 p.i. (P < 0.01). The significant reduction of salmonellae in the caecum was recorded at the end of the experiment (day 14 p.i.) in the EF+SE group in comparison to the SE group (P < 0.01). At day 4 p.i., colonies of S. Enteritidis PT4 were found in the liver of chicks of the SE group in a higher concentration than in chicks of the EF+SE group (P < 0.001). Salmonellae were isolated from the liver until days 8 and 6 p.i. in the SE and EF+SE groups, respectively. The mean values of actual lymphocyte subpopulations in the blood and the relative percentage of caecal intraepithelial lymphocyte subpopulations (CD4, CD8, CD44, TCR, MHC II and IgM) were not influenced at a statistically significant level by the application of the EF 55 and/or the SE PT4 strain. The results demonstrate the antimicrobial effect of E. faecium EF 55 against S. Enteritidis PT4.

Deletion/blockade of Septin 5 converts &amp;ldquo;microdomain&amp;rdquo; to &amp;ldquo;nanodomain&amp;rdquo; coupling between Ca2+ channels and synaptic vesicles at a developing central synapse

Deletion/blockade of Septin 5 converts &amp;ldquo;microdomain&amp;rdquo; to &amp;ldquo;nanodomain&amp;rdquo; coupling between Ca2+ channels and synaptic vesicles at a developing central synapse

Novel Spliced MLL Fusions Have Been Identified Involving the MLL Partner Genes ELL, EPS15, MLLT3, and SEPT5.

Chromosomal rearrangements of the human MLL gene are a genetic hallmark for aggressive acute leukemias. More than 60 partner genes have been characterized at the molecular level until now. The observed recombination mechanisms between MLL and its many recombination partner genes include reciprocal chromosomal translocations, insertions, and intrachromosomal 11q deletions and inversion. “Spliced MLL fusions”, have recently been described as a novel mechanism to generate functional chimeric fusion transcripts for 50% of t(11;19)(q23;p13.3) leukemia patients where the MLL gene is fused to the MLLT1 (ENL) gene and for two single leukemia patients. One patient had a t(11;15) involving the ZFYVE19 gene and the other patient had a 11q interstitial deletion involving the DCPS gene. Within these cases the disrupted MLL gene is fused only on RNA level to a functional transcription unit located in the vicinity of the breakpoint. Here we describe eight novel “spliced MLL fusions” involving the MLL partner genes ELL (1x), EPS15 (3x), MLLT3 (1x), and SEPT5 (1x). For ELL, MLLT3 and SEPT5 the identified spliced fusion seems to be a single or rare event. However 1/3 of the t(1;11)(p32;q23) leukemia cases exhibit “spliced MLL fusions”. I.e. approximately 1/3 of all breakpoints are located upstream of the EPS15 gene. Further studies will help to investigate the remaining question whether these two groups of t(1;11)(p32;q23) leukemia patients have any significant difference in outcome. Interestingly, for the t(11;22)(q23;q11.2) leukemia the major identified fusion transcript has an additional 57 bp exon derived from the 2 kb area up-stream of the SEPT5 gene. Furthermore this patient has a T-ALL and thus differs from all other cases where the patients show myeloid markers when the MLL gene is fused to one of the five members of the septine family. This is leading to the question whether or not this 57 bp big exon can be made responsible for the lineage shift. With these results the number of genes involved in „spliced MLL fusions” has increased from three to seven. Supported in part by grants Ma 1876/7-1, /8-1 and /9-1 from the DFG, grant N1KR-S12T13 from the BMBF and grant 102362 from the Deutsche Krebshilfe.

Phosphorylation of Adult Type Sept5 (CDCrel-1) by Cyclin-dependent Kinase 5 Inhibits Interaction with Syntaxin-1

Increasing evidence implicates cyclin-dependent kinase 5 (Cdk5) in neuronal synaptic function. We searched for Cdk5 substrates in synaptosomal fractions prepared from mouse brains. Mass spectrometric analysis after two-dimensional SDS-PAGE identified several synaptic proteins phosphorylated by Cdk5-p35; one protein identified was Sept5 (CDCrel-1). Although septins were isolated originally as cell division-related proteins in yeast, Sept5 is expressed predominantly in neurons and is implicated in exocytosis. We confirmed that Sept5 is phosphorylated by Cdk5-p35 in vitro and identified Ser17 of adult type Sept5 (Sept5_v1) as a major phosphorylation site. We found that Ser17 of Sept5_v1 is phosphorylated in mouse brains. Coimmunoprecipitation from synaptosomal fractions and glutathione S-transferase-syntaxin-1A pulldown assays of Sept5_v1 expressed in COS-7 cells showed that phosphorylation of Sept5_v1 by Cdk5-p35 decreases the binding to syntaxin-1. These results indicate that the interaction of Sept5 with syntaxin-1 is regulated by the phosphorylation of Sept5_v1 at Ser17 by Cdk5-p35.

Deletion of Two Contiguous Genes, Platelet GPIbβ (Glycoprotein Ibβ) and Septin SEPT5, in a Boy with Bernard-Soulier Syndrome and Developmental Delay: A Possible New Contiguous Gene Syndrome.

Mutations or deletions in the genes for platelet GPIb/V/IX (consisting of GPIbα, GPIbβ, GPV, GPIX) cause the bleeding disorder Bernard-Soulier syndrome. Septins are GTP-binding proteins essential for active membrane movement such as vesicle trafficking. In non-dividing cells (i.e. platelets) septins are implicated in exocytosis. Platelets from a SEPT5 knockout mouse show altered serotonin secretion and platelet aggregation suggesting that SEPT5 is involved in secretion in platelets.We report on a 4 year old boy born to consanguineous parents who developed several life-threatening mucosal bleeding episodes (when he suffered from upper respiratory infections) and showed developmental delay. Giant platelets were present in the blood smear, and ristocetin-induced platelet aggregation was reduced indicating that the boy suffered from Bernard-Soulier syndrome. Immune transmission electron microscopy of the platelets showed reduced expression of GPIb. PCR- and Southern analysis demonstrated that exon 1 and partly exon 2 of GPIbβ was deleted on both alleles. Further experiments showed that the deletion extended 5′ from exon 1 of GPIbβ to include the whole gene coding for SEPT5 located 5′ of GPIbβ. After bone marrow transplantation from a HLA-identical unrelated donor the boy developed no further bleeding symptoms.Conclusions: We identified a patient with Bernard-Soulier syndrome and mental retardation who had not only a homozygous deletion of exon 1 and partly exon 2 of GPIbβ but also a deletion of the whole adjacent SEPT5 gene, possibly indicating a novel contiguous gene syndrome. Whether absence of SEPT5 contributed to particularly severe bleeding symptoms and to developmental delay remains to be proven.

Use of lytic bacteriophages to reduce Salmonella Enteritidis in experimentally contaminated chicken cuts

Reducing Salmonella contamination in poultry is of major importance to prevent the introduction of this microorganism into the food chain. Salmonellae may spread during storage time (shelf life) whenever pre-harvest control fails or post-harvest contamination occurs. Therefore, preventive measures should also be used in the post-harvest level of poultry production in order to control salmonellae. Chicken skin samples were experimentally contaminated by immersing whole legs (thighs and drumsticks) in a suspension containing 10(6) colony forming units per milliliter (CFU/mL) of Salmonella Enteritidis phage type 4 (SE PT4) at the slaughter day. One day later, samples from one group were immersed in a suspension pool containing 10(9) CFU/mL of each of three wild salmonella-lytic bacteriophages previously isolated from feces of free-range chickens. Salmonella counting was performed at three-day intervals in the chicken legs stored at 5°C and showed a significant reduction (P<0.05) of SE PT4 in bacteriophage-treated cuts on days 3, 6 and 9 post-treatment. These findings suggest that the use of bacteriophages may reduce SE PT4 in chicken skin. Further studies are encouraged and might demonstrate the potential of this approach as an efficient and safe technique to be routinelly used for Salmonella control in chicken products.

Genetic deletion of mouse platelet glycoprotein Ibβ produces a Bernard-Soulier phenotype with increased α-granule size

AbstractHere we report the characterization of a mouse model of the Bernard-Soulier syndrome generated by a targeted disruption of the gene encoding the glycoprotein (GP) Ibβ subunit of the GP Ib-IX complex. Similar to a Bernard-Soulier model generated by disruption of the mouse GP Ibα subunit, GP IbβNull mice display macrothrombocytopenia and a severe bleeding phenotype. When examined by transmission electron microscopy, the large platelets produced by a GP IbβNull genotype revealed α-granules with increased size as compared with the α-granules from control mouse platelets. Data are presented linking the overexpression of a septin protein, SEPT5, to the presence of larger α-granules in the GP IbβNull platelet. The SEPT5 gene resides approximately 250 nucleotides 5′ to the GP Ibβ gene and has been associated with modulating exocytosis from neurons and platelets as part of a presynaptic protein complex. Fusion mRNA transcripts present in megakaryocytes can contain both the SEPT5 and GP Ibβ coding sequences as a result in an imperfect polyadenylation signal within the 3′ end of both the human and mouse SEPT5 genes. We observed a 2- to 3-fold increase in SEPT5 protein levels in platelets from GP IbβNull mice. These results implicate SEPT5 levels in the maintenance of normal α-granule size and may explain the variant granules associated with human GP Ibβ mutations and the Bernard-Soulier syndrome.

Salmonella enteritidis, Phage Type 4 Infection in a Commercial Layer Flock in Southern California: Bacteriologic and Epidemiologic Findings

Salmonella enteritidis, phage type 4 (SE PT4), was isolated from five of six 27-wk-old layer chickens submitted for necropsy from a flock of 43,000. Bacteriologic and epidemiologic investigations on the ranch revealed that five of the eight flocks (n = 176,000) were infected. The prevalence of SE PT4 in randomly selected healthy birds ranged from 1.7% (in caged birds) to 50% (in free-range birds) and prevalence in culled birds (kept on dirt floor houses) ranged from 14% to 42%. The estimated overall prevalence of group D Salmonella in eggs contaminated with group D Salmonella was 2.28 per 10,000. The estimated prevalence of group D Salmonella in eggs from caged birds in three infected houses ranged from 1.5 to 4.1 per 10,000, whereas in two houses of free-range birds, prevalence was 14.9 to 19.1 per 10,000. Three of the eight flocks on the ranch remained negative for Salmonella between May 1994 and December 1995 or until removed from the ranch. Salmonella enteritidis PT4 was also isolated from 12.5% (6 of 48) of mice; 57% (four of seven) of cats; and two of two skunks tested. Environmental drag swabs and well water samples yielded multiple serotypes of Salmonella (23/180 and 5/14, respectively) but not S. enteritidis.