Microbes and Oxytocin: Benefits for Host Physiology and Behavior.

It is now understood that gut bacteria exert effects beyond the local boundaries of the gastrointestinal tract to include distant tissues and overall health. Prototype probiotic bacterium Lactobacillus reuteri has been found to upregulate hormone oxytocin and systemic immune responses to achieve a wide array of health benefits involving wound healing, mental health, metabolism, and myoskeletal maintenance. Together these display that the gut microbiome and host animal interact via immune-endocrine-brain signaling networks. Such findings provide novel therapeutic strategies to stimulate powerful homeostatic pathways and genetic programs, stemming from the coevolution of mammals and their microbiome.

Correlation of adherence to the 2012 Infectious Diseases Society of America practice guidelines with patient outcomes in the treatment of diabetic foot infections in an outpatient parenteral antimicrobial programme.

AIM: To evaluate adherence to the 2012 Infectious Diseases Society of America practice guidelines for the management of patients with diabetic foot infections and to determine an association between adherence and clinical outcome. METHODS: A retrospective chart review was performed to evaluate the management and clinical outcomes of patients with diabetic foot infections treated with outpatient parenteral antimicrobial therapy between 1 January 2011 and 30 June 2012 at Wishard Health Services/Eskenazi Health. Adherence to individual Infectious Diseases Society of America diabetic foot infection treatment guideline recommendations was measured, and then assessed in relation to clinical outcome. RESULTS: A total of 57 patients (61% male, mean age 54 years) with moderate to severe diabetic foot infection met the inclusion criteria. None of the treatment courses of these patients adhered to all the Infectious Diseases Society of America guideline recommendations. The recommendations most frequently adhered to were consultation of appropriate multidisciplinary teams (n=54, 94.7%) and performance of diagnostic imaging (n=52, 89.5%). The recommendations least frequently adhered to were diabetic foot wound classification scoring on admission (n=0, 0%), appropriate culture acquisition (n=12, 21.2%), surgical intervention when indicated (n=32, 46.2%) and appropriate empiric antibiotic selection (n=34, 59.7%). Of 56 patients, 52 (92.9%) experienced clinical cure at the end of outpatient parenteral antimicrobial therapy compared with 34 of 53 patients (64%) at 6 months after the completion of therapy. Adherence to individual guidelines was not associated with clinical outcome. Patients who experienced treatment failure were more likely to have severe diabetic foot infection or peripheral neuropathy. CONCLUSIONS: Adherence to the Infectious Diseases Society of America diabetic foot infection guideline recommendations was found to be suboptimal in the present study. The effect of adhering to individual Infectious Diseases Society of America diabetic foot infection recommendations on clinical outcome needs to be investigated.

Probiotic 'glow of health': it's more than skin deep.

Radiant skin and hair are universal indicators of good health. It was recently shown that feeding of probiotic bacteria to aged mice rapidly induced youthful vitality characterised by thick lustrous skin and hair, and enhanced reproductive fitness, not seen in untreated controls. Probiotic-treated animals displayed integrated immune and hypothalamic-pituitary outputs that were isolated mechanistically to microbe-induced anti-inflammatory interleukin-10 and neuropeptide hormone oxytocin. In this way, probiotic microbes interface with mammalian physiological underpinnings to impart superb physical and reproductive fitness displayed as radiant and resilient skin and mucosae, unveiling novel strategies for integumentary health.

Helicobacter hepaticus infection in mice: models for understanding lower bowel inflammation and cancer.

Pioneering work in the 1990s first linked a novel microaerobic bacterium, Helicobacter hepaticus, with chronic active hepatitis and inflammatory bowel disease in several murine models. Targeted H. hepaticus infection experiments subsequently demonstrated its ability to induce colitis, colorectal cancer, and extraintestinal diseases in a number of mouse strains with defects in immune function and/or regulation. H. hepaticus is now widely utilized as a model system to dissect how intestinal microbiota interact with the host to produce both inflammatory and tolerogenic responses. This model has been used to make important advances in understanding factors that regulate both acquired and innate immune response within the intestine. Further, it has been an effective tool to help define the function of regulatory T cells, including their ability to directly inhibit the innate inflammatory response to gut microbiota. The complete genomic sequence of H. hepaticus has advanced the identification of several virulence factors and aided in the elucidation of H. hepaticus pathogenesis. Delineating targets of H. hepaticus virulence factors could facilitate novel approaches to treating microbially induced lower bowel inflammatory diseases.

Nonsteroidal anti-inflammatory drug hypersensitivity syndrome. A multicenter study. I. Clinical findings and in vitro diagnosis.

BACKGROUND: We present the results obtained from the largest series of in vitro diagnostic tests ever reported in patients with clinically validated hypersensitivity to acetylsalicylic acid (ASA)/nonsteroidal anti-inflammatory drugs (NSAID) compared with various categories of controls tolerating ASA/NSAIDs. This multicenter study, which was performed within the framework of the European Network for Drug Allergy (ENDA) group, showed that the basophil activation test (BAT), particularly when used with the 3 NSAIDs aspirin (ASA), diclofenac (DIC), and naproxen (NAP), allows us to confirm the diagnosis of NSAID hypersensitivity syndrome. The results of the cellular allergen stimulation test (CAST) frequently correlate with those of the BAT, although not always. An unexpected finding was that basophil activation by NSAIDs is not an all-or-nothing phenomenon restricted to clinically hypersensitive patients, but that it also occurs in a dose-related manner in some NSAID-tolerant control individuals.Therefore, NSAID hypersensitivity appears as a shift in the normal pharmacological response to NSAIDs. These findings allow us to formulate a new rational hypothesis about the mechanism of NSAID hypersensitivity syndrome, a mechanism that most authors continue to describe as "unknown." METHODS: We enrolled 152 patients with a history of hypersensitivity to NSAIDs and 136 control participants in 11 different centers between spring 2003 and spring 2006. Flowcytometric BAT was performed. RESULTS: The most noteworthy results of our study were that 57% of 140 patients presented very clear-cut positive BAT results to multiple NSAIDs, and 16% were entirely negative. In about 27% of cases, positive results were obtained with 1 or 2 concentrations of a single NSAID. There is clearly a correlation between the results of BAT and CAST. CONCLUSIONS: BAT seems particularly indicated in patients with a clinical history of NSAID intolerance, and in whom a provocation test is not advisable for ethical, clinical, or other reasons. Clear-cut positive results can be considered as confirming a history of NSAID hypersensitivity, although negative results may not exclude it.

Nitric oxide and TNF-alpha trigger colonic inflammation and carcinogenesis in Helicobacter hepaticus-infected, Rag2-deficient mice.

Recombinase-activating gene-2-deficient (Rag2(-/-)) mice lacking functional lymphocytes provide a useful model of chronic inflammatory bowel disease-emulating events in human colon cancer. Infection of Rag2(-/-) mice with Helicobacter hepaticus led to accumulation of macrophages and neutrophils in the colon, a process temporally related to up-regulation of tissue inducible nitric oxide synthase (iNOS) expression at the site of infection and increased nitric oxide (NO) production, as evidenced by urinary excretion of nitrate. Progressive development of increasingly severe inflammation, hyperplasia, dysplasia, and cancer accompanied these changes. Concurrent administration of an iNOS inhibitor prevented NO production and abrogated epithelial pathology and inhibited the onset of cancer. The presence of Gr-1(+) neutrophils and elevated tumor necrosis factor-alpha (TNF-alpha) expression in colon were required for increased iNOS expression and cancer, whereas interleukin-10 (IL-10) down-regulated TNF-alpha and iNOS expression and suppressed cancer. Anti-inflammatory CD4(+) regulatory lymphocytes also down-regulated iNOS and reduced cancer formation. Collectively, these results confirm essential roles for inflammation, increased TNF-alpha expression, and elevated NO production in colon carcinogenesis.

Hepatobiliary inflammation, neoplasia, and argyrophilic bacteria in a ferret colony.

Hepatobiliary disease was diagnosed in eight of 34 genetically unrelated cohabitating pet ferrets (Mustela putorios furo) during a 7-year period. The eight ferrets ranged in age from 5 to 8 years and exhibited chronic cholangiohepatitis coupled with cellular proliferation ranging from hyperplasia to frank neoplasia. Spiral-shaped argyrophilic bacteria were demonstrated in livers of three ferrets, including two with carcinoma. Sequence analysis of a 400-base pair polymerase chain reaction product amplified from DNA derived from fecal bacteria from one ferret demonstrated 98% and 97% similarity to Helicobacter cholecystus and Helicobacter sp. strain 266-1 , respectively. The clustering of severe hepatic disease in these cohabitating ferroes suggests a possible infectious etiology. The role of Helicobacter species and other bacteria in hepatitis and/or neoplasia in ferrets requires further study.

A filamentous growth response mediated by the yeast mating pathway.

Haploid cells of the budding yeast Saccharomyces cerevisiae respond to mating pheromones by arresting their cell-division cycle in G1 and differentiating into a cell type capable of locating and fusing with mating partners. Yeast cells undergo chemotactic cell surface growth when pheromones are present above a threshold level for morphogenesis; however, the morphogenetic responses of cells to levels of pheromone below this threshold have not been systematically explored. Here we show that MATa haploid cells exposed to low levels of the alpha-factor mating pheromone undergo a novel cellular response: cells modulate their division patterns and cell shape, forming colonies composed of filamentous chains of cells. Time-lapse analysis of filament formation shows that its dynamics are distinct from that of pseudohyphal growth; during pheromone-induced filament formation, daughter cells are delayed relative to mother cells with respect to the timing of bud emergence. Filament formation requires the RSR1(BUD1), BUD8, SLK1/BCK1, and SPA2 genes and many elements of the STE11/STE7 MAP kinase pathway; this response is also independent of FAR1, a gene involved in orienting cell polarization during the mating response. We suggest that mating yeast cells undergo a complex response to low levels of pheromone that may enhance the ability of cells to search for mating partners through the modification of cell shape and alteration of cell-division patterns.

Typhlocolitis in NF-kappa B-deficient mice.

Activation of inflammatory gene expression by the transcription factor NF-kappaB is a central pathway in many inflammatory disorders, including colitis. Increased NF-kappaB activity has been linked with development of colitis in humans and animal models, thus it was unexpected when NF-kappaB-deficient mice developed spontaneous typhlocolitis. To further characterize this finding, we induced typhlocolitis in rederived NF-kappaB-deficient mice using intragastric infection with Helicobacter hepaticus. At 6 wk postinfection (PI), severe colitis with increased type 1 cytokine expression was seen in infected mice that lacked the p50 subunit of NF-kappaB and were also heterozygous for the p65 subunit of NF-kappaB(p50(-/-)p65(+/-)). Mice lacking the p50 subunit alone (p50(-/-)) were less severely affected, and wild-type mice and p65(+/-) mice were unaffected. T cell development in NF-kappaB-deficient mice was normal. These data indicate that p50 and p65 subunits of NF-kappaB have an unexpected role in inhibiting the development of colitis.

Gastric dilatation syndrome associated with chronic nephropathy, hypergastrinemia, and gastritis in mice exposed to high levels of environmental antigens.

Gastric dilatation (GD) has been observed in Tac:(SW)fBR surveillance mice, with mean age of 10 months, that are exposed to high levels of environmental antigens during routine exposure to dirty bedding. The aim of the study reported here was to determine whether GD was associated with other systemic conditions affecting mice. Three groups of nine animals including-surveillance mice not exposed to dirty bedding (control), surveillance mice with out GD (NGD), and surveillance mice with GD (group GD)-had mean stomach weight with ingesta of 0.5 +/- 0.02 g, 1.09 +/- 0.07 g (P < 0.0001), and 2.54 +/- 0.4 g (P < 0.0001), respectively. Mean serum creatinine concentration was significantly higher in GD (1.6 +/- 0.25 mg/dl), compared with NGD (0.17 +/- 0.22 mg/dl, P < 0.0001) and control (0.2 +/- 0.16 mg/ dl, P < 0.0001) mice. In addition, lesions consistent with severe chronic nephropathy and mild gastritis were common in GD, compared with NGD and control mice. Finally, serum amidated gastrin concentration was significantly high in GD (179.37 +/- 53.86 pM, P < 0.03) and NGD (264.89 +/- 115.89 pM, P < 0.009), compared with control (60.77 +/- 8.39 pM) mice. Gastric dilatation syndrome is associated with chronic nephropathy, hypergastrinemia, and gastritis in surveillance mice exposed to high levels of environmental antigens.

Café-au-lait spots and early onset colorectal neoplasia: a variant of HNPCC?

BACKGROUND: Café-au-lait spots (CALS) are classically found in neurocutaneous syndromes such as neurofibromatosis, but have not been associated with hereditary colorectal cancer. However, review of hereditary colorectal cancer case reports reveals occasional description of CALS on physical exam. METHODS: We describe the colonic and extracolonic phenotype in a family with CALS and early onset colorectal neoplasia (adenomas and/or cancer) and review 23 additional families reported in the literature. RESULTS: Among the 24 families, 32/59 (54.2%) individuals had colorectal adenomas diagnosed at a mean age of 15.7 +/- 1.1 (SE) years (range 5-38 years). The majority (24/32, 75.0%) of persons at first colorectal examination had oligopolyposis (< 100 polyps) versus polyposis (> or = 100 polyps). Forty-two of 59 (71.2%) individuals were affected with colorectal cancer, diagnosed at a mean age of 31.9 +/- 2.7 years (range 5-70 years). A brain tumor was found in 28/59 (47.5%) affected individuals (4 families with 2 or more cases) with an overall mean age of diagnosis of 16.5 +/- 1.2. Lymphoma and/or leukemia was found in 8/24 (33.3%) families (one family with 3 cases). Two families had mutation of the mismatch repair gene, hPMS2 (1 with homozygous germline mutation), while two carried homozygous germline mutations of another mismatch repair gene, hMLH1. CONCLUSIONS: Café-au-lait spots with early onset colorectal neoplasia may identify families with a variant of HNPCC characterized by oligopolyposis, glioblastoma at young age, and lymphoma. This variant may be caused by homozygous mutation of the mismatch repair genes, such as hPMS2 or hMLH1.

E2F4 is essential for normal erythrocyte maturation and neonatal viability.

The retinoblastoma protein (pRB) plays a key role in the control of normal development and proliferation through the regulation of the E2F transcription factors. We generated a mutant mouse model to assess the in vivo role of the predominant E2F family member, E2F4. Remarkably, loss of E2F4 had no detectable effect on either cell cycle arrest or proliferation. However, E2F4 was essential for normal development. E2f4-/- mice died of an increased susceptibility to opportunistic infections that appeared to result from craniofacial defects. They also displayed a variety of erythroid abnormalities that arose from a cell autonomous defect in late stage maturation. This suggests that E2F4 makes a major contribution to the control of erythrocyte development by the pRB tumor suppressor.

APC-dependent changes in expression of genes influencing polyamine metabolism, and consequences for gastrointestinal carcinogenesis, in the Min mouse.

The colorectal mucosa of pre-symptomatic individuals with familial adenomatous polyposis (FAP) contains elevated levels of the proliferation-associated polyamines. The Min mouse, like humans with FAP, expresses an abnormal genotype for the APC tumor suppressor gene. In order to determine how APC mutation influences intestinal tissue polyamine content, we measured steady-state RNA levels of ornithine decarboxylase (ODC), the first enzyme in polyamine synthesis, antizyme (AZ), a protein which negatively regulates ODC, and the spermidine/spermine N(1)-acetyltransferase (SSAT), the first enzyme in polyamine catabolism. RNA content was increased 6- to 8-fold in both the small intestine and colon for ODC, decreased significantly in the small intestine but not the colon for AZ and was not statistically different in either intestinal tissue for SSAT in Min mice compared with normal littermates. Consistent with the changes in ODC and AZ gene expression, small intestinal, but not colonic, polyamine content was elevated in Min mice compared with normal littermates. Treatment of Min mice with the specific ODC inhibitor difluoromethylornithine (DFMO) suppressed small intestinal, but not colonic, polyamine content and tumor number. These data indicate that small intestinal tissue polyamine content is elevated in Min mice by a mechanism involving APC-dependent changes in ODC and AZ RNA. Further, ODC enzyme activity, which is influenced by both ODC and AZ RNA levels and inhibited by DFMO, is consequential for small intestinal tumorigenesis in this model. In the FAP population, DFMO may be of value in the chemoprevention of small intestinal adenocarcinoma that remains a risk following colectomy.

Pregnancy toxemia in the European ferret (Mustela putorius furo).

BACKGROUND AND OBJECTIVE: Pregnancy toxemia may lead to appreciable mortality among jills and their offspring. The objective of this report was to increase awareness of the disease, its likely cause, and practical prevention and treatment measures. METHODS: Ten cases of pregnancy toxemia were evaluated. Jills were in late gestation (mean, 38 days; range, 34 to 42 days) and had large litters (mean, 11.5 kits; range, 7 to 15 kits). RESULTS: The most common clinical signs of disease were lethargy, inappetence, dehydration, and excess shedding. Hematologic and clinical biochemical abnormalities included anemia (4 of 8 jills tested), hypoproteinemia (5 of 7), azotemia (7 of 7), hypocalcemia (5 of 6), hyperbilirubinemia (3 of 3), and high liver enzyme activities (6 of 6). Two jills were found dead; two jills were euthanized, six received supportive treatment, and cesarean section was performed on five. The three jills that survived tended to have less pronounced azotemia, hypoproteinemia, and liver enzyme activity increases and were not anemic. Hepatic lipidosis was observed grossly in all jills that died and was confirmed by histologic examination in four jills. CONCLUSIONS: Pregnancy toxemia in ferrets resembles metabolic diseases in several other animal species and requires aggressive treatment, including supportive care, nutritional supplementation, and cesarean section. Maintaining adequate nutrition and avoiding stress late in gestation may prevent the disease.

Effects of N-methyl-D-aspartate (dizocilpine) and alpha-amino-3-hydroxy-4-isoxazolepropionate (LY215490) receptor antagonists on the voiding reflex induced by perineal stimulation in the neonatal rat.

The present study was undertaken to examine the role of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate and N-methyl-D-aspartate glutamate receptors in the regulation of voiding reflexes induced by perineal stimulation in the neonatal rat. Four-, six- and 10-day-old awake rats were used in the experiments and perineal stimulation was applied using the tip of a 1-ml syringe to evoke voiding. Voided volume and residual volume were measured. In 10-day-old rats, LY215490 (3-10 mg/kg, i.p.), a competitive alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptor antagonist, significantly inhibited reflex voiding, increasing the residual volume 34-53-fold. A 3 mg/kg dose decreased the urine release by 55%, whereas 10 mg/kg totally suppressed the voiding reflex induced by the perineal stimulation. LY215490 (10 mg/kg, i.p.) produced similar effects in four- and six-day-old rats. Dizocilpine (1-3 mg/kg, i.p.), a non-competitive N-methyl-D-aspartate receptor antagonist, also significantly decreased the urine release (62-82%) and increased residual volume (180-230-fold). Combined administration of LY215490 (1 mg/kg, i.p.) and dizocilpine (0.3 mg/kg, i.p.) to 10-day-old rats, in doses that individually had no effect on perineal stimulation-evoked voiding, depressed voided volume by 65%. These results indicate that, in neonatal rats, glutamatergic transmission in the spinal cord has an essential role in reflex micturition induced by perineal stimulation, and that facilitatory interactions between alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate and N-methyl-D-aspartate glutamatergic mechanisms are important for voiding, as noted previously in adult rats.

Nutritional imperatives in cystic fibrosis therapy.

Patients with CF who receive optimal nutrition have better growth, maintain better nutritional reserves, and have better pulmonary function than patients with CF who have poor nutrition. These factors influence quality of life as well as survival. The metabolic and immunologic response to infection, as well as the increased work of breathing, escalates calorie requirements in this patient population. No single strategy works for every patient. Therefore, close monitoring of growth, symptoms, and changes in respiratory status that could increase calorie requirements is necessary.

Immunophenotypic characterization of lymphomas from the mediastinum of young ferrets.

OBJECTIVE: To determine the phenotype of naturally developing lymphomas in young ferrets. ANIMALS: 10 ferrets with lymphoma. PROCEDURE: Neoplastic tissues were graded histologically according to the National Cancer Institute's Working Formulation for non-Hodgkin's lymphoma and phenotype was determined by means of immunohistochemical staining. A polyclonal anti-human CD3 and a monoclonal anti-human CD79 antibody were used to classify the lymphomas in situ as T-cell or B-cell origin. Specificity of antibodies was determined by evaluating lymphoid tissue from normal ferrets in situ, which was confirmed by western blot analyses. RESULTS: All 10 ferrets had clinically aggressive tumors, irrespective of the phenotype. Nine ferrets had T-cell lymphoma that extensively involved the mediastinum. Remnants of thymic tissue, indicative of thymic origin, were identified in lymphoma of these 9 ferrets. One ferret had a B-cell multicentric lymphoma without involvement of the mediastinum. CONCLUSIONS: The majority of lymphomas in these young ferrets involved the mediastinum and were of T-cell phenotype. Impact for Human Medicine-There are many similarities between the lymphoma syndrome of ferrets and the condition documented for cats and children with lymphoma of the mediastinal area. CLINICAL RELEVANCE: Differential diagnoses for young ferrets with clinical signs of lethargy or respiratory distress should include T-cell lymphoma of the mediastinum.

Pheromone-regulated genes required for yeast mating differentiation.

Yeast cells mate by an inducible pathway that involves agglutination, mating projection formation, cell fusion, and nuclear fusion. To obtain insight into the mating differentiation of Saccharomyces cerevisiae, we carried out a large-scale transposon tagging screen to identify genes whose expression is regulated by mating pheromone. 91,200 transformants containing random lacZ insertions were screened for beta-galactosidase (beta-gal) expression in the presence and absence of alpha factor, and 189 strains containing pheromone-regulated lacZ insertions were identified. Transposon insertion alleles corresponding to 20 genes that are novel or had not previously been known to be pheromone regulated were examined for effects on the mating process. Mutations in four novel genes, FIG1, FIG2, KAR5/ FIG3, and FIG4 were found to cause mating defects. Three of the proteins encoded by these genes, Fig1p, Fig2p, and Fig4p, are dispensible for cell polarization in uniform concentrations of mating pheromone, but are required for normal cell polarization in mating mixtures, conditions that involve cell-cell communication. Fig1p and Fig2p are also important for cell fusion and conjugation bridge shape, respectively. The fourth protein, Kar5p/Fig3p, is required for nuclear fusion. Fig1p and Fig2p are likely to act at the cell surface as Fig1:: beta-gal and Fig2::beta-gal fusion proteins localize to the periphery of mating cells. Fig4p is a member of a family of eukaryotic proteins that contain a domain homologous to the yeast Sac1p. Our results indicate that a variety of novel genes are expressed specifically during mating differentiation to mediate proper cell morphogenesis, cell fusion, and other steps of the mating process.

Adjustment to hearing impairment II: audiological and demographic correlates.

To study adjustment to hearing impairment, clinical records from a five-center consortium (N = 1,008) were used to create a heterogeneous clinical database with results of audiometric tests, demographic and case history information, and responses to the Communication Profile for the Hearing Impaired (CPHI; Demorest & Erdman, 1986, 1987). Normative findings have been described previously (Erdman & Demorest, 1998). Hierarchical regression analyses revealed that audiometric variables were moderately correlated with communication performance, behavioral strategies, and personal adjustment. With hearing impairment controlled statistically, age and education effects were evident in many areas of adjustment; correlations between adjustment and gender were relatively weak; and marital status, employment status, and race/ethnicity were rarely significant correlates.