Small RNA populations reflect the complex dialogue established between heterograft partners in grapevine.

Grafting is an ancient method that has been intensively used for the clonal propagation of vegetables and woody trees. Despite its importance in agriculture the physiological and molecular mechanisms underlying phenotypic changes of plants following grafting are still poorly understood. In the present study, we analyse the populations of small RNAs in homo and heterografts and take advantage of the sequence differences in the genomes of heterograft partners to analyse the possible exchange of small RNAs. We demonstrate that the type of grafting per se dramatically influences the small RNA populations independently of genotypes but also show genotype specific effects. In addition, we demonstrate that bilateral exchanges of small RNAs, mainly short interfering RNAs, may occur in heterograft with the preferential transfer of small RNAs from the scion to the rootstock. Altogether, the results suggest that small RNAs may have an important role in the phenotype modifications observed in heterografts.

Epidemiology and risk factors for extrapulmonary tuberculosis in Lebanon.

SETTING: Lebanon is a relatively low TB-burden country, but has a high proportion of extrapulmonary tuberculosis (EPTB). Concern has been expressed that Syrian-born refugees could add to the TB burden and rates of EPTB: since 2011, >1 000 000 Syrian refugees have entered Lebanon.OBJECTIVE: The Lebanese National Tuberculosis Programme (NTP) sought to identify factors for the high proportion of EPTB and to assess the potential impact of Syrian refugees.DESIGN: NTP line-listed data from 2014-2015 were analyzed with logistic regression identified risk factors for EPTB. A trend analysis for 2011-2015 assessed TB burden by nationality and site of TB.RESULTS: Of 1347 reported TB cases from 2014 to 2015, 507 (38%) were EPTB and 46% were Lebanese. In analysis limited to Lebanese-born, the proportion of EPTB cases was relatively stable, 47% in 2011 and 52% in 2015. Modeling identified risk factors for EPTB as being female (aOR 1.79, 95%CI 1.39-2.32) and 5-15 years old (aOR 3.31, 95%CI 1.47-7.45) compared with children aged <5 years. Between 2011 and 2015, the proportion of TB cases among Syrian-born increased from 3% in 2011 to 21% in 2015 (P < 0.001); however, the proportion of EPTB versus PTB cases among Syrians remained stable (P = 0.264).CONCLUSION: Syrian TB cases increased almost 10-fold in five years (2011-2015) but their contribution to EPTB did not change. The high proportion of EPTB in Lebanon and those aged 5-15 years merits further investigation.

Prevalence of mental disorders and epidemiological associations in post-conflict primary care attendees: a cross-sectional study in the Northern Province of Sri Lanka.

BACKGROUND: Experiencing conflict and displacement can have a negative impact on an individual's mental health. Currently, prevalence of mental health disorders (MHDs) at the primary care level in post-conflict areas within the Northern Province of Sri Lanka is unknown. We aimed to explore this prevalence in conflict-affected populations attending primary care, using a structured package of validated screening tools for MHDs. METHODS: This cross-sectional study aimed to determine factors related to mental health disorders at the primary care level in Northern Province, Sri Lanka. A structured interview was conducted with internally displaced adults attending 25 randomly selected primary care facilities across all districts of Northern Sri Lanka (Jaffna, Mannar, Mullaitivu, Vavuniya). Participants were screened for depression, anxiety, psychosis, PTSD, and somatoform symptoms. RESULTS: Among 533 female and 482 male participants (mean age 53.2 years), the prevalence rate for any MHD was 58.8% (95% CI, 53.8-61.4), with 42.4% screening positive for two or more disorders (95% CI, 38.6-46.1). Anxiety prevalence was reported at 46.7% (95% CI, 41.9-51.5), depression at 41.1% (95% CI, 38.7-44.5), PTSD at 13.7% (95% CI, 10.6-16.8), somatoform symptoms at 27.6% (95% CI, 23.6-31.5), and psychosis with hypomania at 17.6% (95% CI, 13.3-21.9). CONCLUSION: This is the first study at the primary care level to investigate prevalence of MHDs among conflict-affected populations in the Northern Province, Sri Lanka. Results highlight unmet mental health needs in the region. Training intervention to integrate mental health services into primary care is planned.

Heterografting with nonself rootstocks induces genes involved in stress responses at the graft interface when compared with autografted controls.

Although grafting is widely used in the agriculture of fruit-bearing crops, little is known about graft union formation in particular when two different species are grafted together. It is fascinating that two different plant species brought together can develop harmoniously as one organism for many decades. The objective of this study was to determine whether grafting two different grapevine genotypes alters gene expression at the graft interface in comparison to the presumably wound-like gene expression changes induced in autografts. Gene expression at the graft interface was studied 3, 7, 14, and 28 d after grafting in hetero- and autografts of grapevine (Vitis spp.). Genes differentially expressed between the hetero- and autografts during graft union formation were identified. These genes were clustered according to their expression profile over the time course. MapMan and Gene Ontology enrichment analysis revealed the coordinated upregulation of genes from numerous functional categories related to stress responses in the hetero- compared to the autografts. This indicates that heterografting with nonself rootstocks upregulates stress responses at the graft interface, potentially suggesting that the cells of the graft interface can detect the presence of a nonself grafting partner.

Determining TB rates and TB case burden for refugees.

SETTING: DeKalb County, Georgia. OBJECTIVES: To calculate and compare tuberculosis (TB) rates in refugees to US-born, total foreign-born (refugee and other), and other foreign-born persons and to determine the contribution of refugees to the county TB case burden. METHODS: The study included: (1) collection of county TB case numbers and population figures from 1995 through 1999; (2) estimation of the refugee population; (3) comparison of TB rates; and (4) calculation of the refugee TB case burden. Sensitivity analysis was performed on refugee population estimates. RESULTS: From 1995 through 1999, estimating that refugees made up 10% of the foreign-born population, the average TB rate for refugees was 83.2 per 100,000, compared with 12.7 for US-born persons. From 1997 through 1999, refugees had a seven-fold greater risk of having TB than US-born persons and a two-fold greater risk than other foreign-born persons. Refugees represented respectively 7.6% and 19.3% of the county and foreign-born TB case burdens. For TB rates to be equal among all foreign-born persons, refugees would need to make up 15-25% of the foreign-born population. CONCLUSION: Despite overseas screening, refugees have high TB rates, and contribute substantially to the county TB case burden. Enhanced surveillance and targeted programs to address TB in refugees should be a public health priority.

The use of microelectrodes to investigate compartmentation and the transport of metabolized inorganic ions in plants.

Microelectrode measurements can be used to investigate both the intracellular pools of ions and membrane transport processes of single living cells. Microelectrodes can report these processes in the surface layers of root and leaf cells of intact plants. By careful manipulation of the plant, a minimum of disruption is produced and therefore the information obtained from these measurements most probably represents the 'in vivo' situation. Microelectrodes can be used to assay for the activity of particular transport systems in the plasma membrane of cells. Compartmental concentrations of inorganic metabolite ions have been measured by several different methods and the results obtained for the cytosol are compared. Ion-selective microelectrodes have been used to measure the activities of ions in the apoplast, cytosol and vacuole of single cells. New sensors for these microelectrodes are being produced which offer lower detection limits and the opportunity to measure other previously unmeasured ions. Measurements can be used to determine the intracellular steady-state activities or report the response of cells to environmental changes.

The HeartSmart Family Fun Pack: an evaluation of family-based intervention for cardiovascular risk reduction in children.

In 1998, the Heart and Stroke Foundation launched the HeartSmart Family Fun Pack, a "user-friendly" resource to support family-based lifestyle changes among children age 6-12 years. Of 1,387 parents who completed a pre-intervention questionnaire, 300 (21%) were surveyed three months later. Comparisons of the pre- and post-intervention surveys show that the Family Fun Pack was effective in supporting healthy lifestyle changes (38% and 28% of families reported improvement in their child's nutrition and level of physical activity, respectively; 12% reported reduced passive smoking). Effectiveness was greatest among those families who self-identified themselves as being in what corresponds to the contemplation, preparation and action stages of change. This evaluation suggests that a relatively low-cost intervention can be effective in supporting positive lifestyle changes if targeted to families at specific points in the stages of change.

Malaria, intestinal parasites, and schistosomiasis among Barawan Somali refugees resettling to the United States: a strategy to reduce morbidity and decrease the risk of imported infections.

In 1997, enhanced health assessments were performed for 390 (10%) of approximately 4,000 Barawan refugees resettling to the United States. Of the refugees who received enhanced assessments, 26 (7%) had malaria parasitemia and 128 (38%) had intestinal parasites, while only 2 (2%) had Schistosoma haematobium eggs in the urine. Mass therapy for malaria (a single oral dose of 25 mg/kg of sulfadoxine-pyrimethamine) was given to all Barawan refugees 1-2 days before resettlement. Refugees >2 years of age and nonpregnant women received a single oral dose of 600 mg albendazole for intestinal parasite therapy. If mass therapy had not been provided, upon arrival in the United States an estimated 280 (7%) refugees would have had malaria infections and 1,500 (38%) would have had intestinal parasites. We conclude that enhanced health assessments provided rapid on-site assessment of parasite prevalence and helped decrease morbidity among Barawan refugees, as well as, the risk of imported infections.

Reduction in cytokine production in colorectal cancer patients: association with stage and reversal by resection.

The aim of this study was to assess monocyte/macrophage function, as defined by lipopolysaccharide (LPS)-induced production of tumour necrosis factor (TNF)-alpha, interleukin (IL)-10 and interferon (IFN)-gamma by stimulated whole blood cultures in patients with colorectal carcinoma before and after surgical resection. Forty colorectal cancer patients prior to surgery and 31 healthy controls were studied. Heparinized venous blood was taken from colorectal cancer patients prior to surgery and from healthy controls. Serial samples were obtained at least 3-6 weeks post-operatively. Blood was stimulated with LPS for 24 h and supernatants were assayed for TNF-alpha, IFN-gamma and IL-10 by enzyme-linked immunosorbent assay. LPS-induced production of TNF-alpha and of IFN-gamma was reduced in patients with colorectal carcinoma compared to controls (TNF-alpha, 11,269 pg/ml(-1) ¿12,598¿; IFN-gamma, 0.00 pg/ml(-1) ¿226¿; median ¿IQR¿) (TNF-alpha, 20,576 pg/m(-1) ¿11,637¿, P < 0.0001; IFN-gamma, 1,048 ¿2,428¿, P = 0.0051, Mann-Whitney U-test). Production in patients after surgery had increased (TNF-alpha: 17,620 pg/ml(-1) ¿7,986¿; IFN-gamma. 410 pg/ml(-1) ¿2,696¿; mean ¿s.d.¿) and were no longer significantly reduced when compared to controls (TNF-alpha, P = 0.28; IFN-gamma, P = 0.76). Production of TNF-alpha and IFN-gamma prior to surgery were reduced to a greater extent in patients with Dukes' stage C tumours compared to those with Dukes' stage A and B stage. There was no difference in IL-10 production between any group. Monocytes/macrophages from patients with colorectal carcinoma are refractory to LPS stimulation as reflected by reduction in TNF-alpha and IFN-gamma production and this is more pronounced in patients with advanced stage tumours. This suppression is not mediated by IL-10 and disappears following surgical resection of the tumour. This provides evidence for tumour induced suppression of immune function in patients with colorectal cancer and identifies a potential therapeutic avenue.

Frequency and nature of cytokine gene polymorphisms in type 1 autoimmune hepatitis.

Genetic involvement in type 1 autoimmune hepatitis (AIH) is indicated by a marked female preponderance and strong, well-established, human leukocyte antigen (HLA) associations. These associations, however, are not universal and a number of genes outside the major histocompatibility complex may also play a role in susceptibility to type 1 AIH. Prime candidates at present are those polymorphic genes encoding the proinflammatory and immunoregulatory cytokines. The aim of this study was to investigate, for the first time, 2 members of the interleukin-1 (IL-1) family (IL-1B and IL-1RN), 3 polymorphic sites in the interleukin-10 (IL-10) gene promoter (positions -1082, -819, and -592), and 2 polymorphisms in the tumor necrosis factor-alpha (TNF-alpha) promoter (positions -308 and -238) in type 1 AIH. The study was performed on 2 independently collected DNA banks, each with appropriate controls, and throughout the analysis associations described in the first set were confirmed in the second set. Standard polymerase chain reaction (PCR)-based genotyping techniques were used. Overall there were no significant differences in the distributions of the IL-1B and IL-10 alleles, genotypes, or haplotypes in either study set. In contrast we report a significant association between type 1 AIH and TNF*2 (first set: 34% of controls vs. 49% of patients, Pc =.014 and second set: 26% vs. 56%, P =.00008). However, TNF*2 is found in strong linkage disequilibrium with the HLA A1-B8-DR3 haplotype and stratification analysis indicates that the association with TNF*2 is interdependent with HLA DRB1*0301. This is an indication that there is more than one susceptibility allele for type 1 AIH on chromosome 6p21.3.

Prospective evaluation of risk factors for bloodstream infection in patients receiving home infusion therapy.

BACKGROUND: Intravenous therapy in the outpatient and home settings is commonplace for many diseases and nutritional disorders. Few data are available on the rate of and risk factors for bloodstream infection among patients receiving such therapy. OBJECTIVE: To determine rates of and risk factors for bloodstream infection among patients receiving home infusion therapy. DESIGN: Prospective, observational cohort study. SETTING: Cleveland, Ohio, and Toronto, Ontario, Canada. PATIENTS: Patients receiving home infusion therapy through a central or midline catheter. MEASUREMENTS: Primary laboratory-confirmed bloodstream infection. RESULTS: Among 827 patients (988 catheters), the most common diagnoses were infections other than HIV (67%), cancer (24%), nutritional and digestive disease (17%), heart disease (14%), receipt of bone marrow or solid organ transplants (11%), and HIV infection (7%). Sixty-nine bloodstream infections occurred during 69,532 catheter-days (0.99 infections per 1000 days). In a Cox regression model with time-dependent covariates, independent risk factors for bloodstream infection were recent receipt of a bone marrow transplant (hazard ratio, 5.8 [95% CI, 3.0 to 11.3]), receipt of total parenteral nutrition (hazard ratio, 4.1 [CI, 2.3 to 7.2]), receipt of therapy outside the home (for example, in an outpatient clinic or physician's office) (hazard ratio, 3.6 [CI, 2.2 to 5.9]), use of a multilumen catheter (hazard ratio, 2.8 [CI, 1.7 to 4.7]), and previous bloodstream infection (hazard ratio, 2.5 [CI, 1.5 to 4.2]). Rates of bloodstream infection per 1000 catheter-days varied from 0.16 for patients with none of these 5 risk factors to 6.77 for patients with 3 or more risk factors. Centrally inserted venous catheters were associated with a higher risk than implanted ports were, but the difference was not statistically significant. CONCLUSION: Bloodstream infections seem to be infrequent among outpatients receiving infusions through central and midline catheters. However, the rate of infection increases with bone marrow transplantation, parenteral nutrition, infusion therapy in a hospital clinic or physician's office, and use of multilumen catheters. Compared with implanted ports or peripherally inserted catheters, centrally inserted venous catheters may confer greater risk for bloodstream infection.

Cytokine polymorphisms associated with clinical features and treatment outcome in type 1 autoimmune hepatitis.

BACKGROUND & AIMS: Polymorphisms that control cytokine production can affect immunoregulation. The frequency and consequences of these polymorphisms in type 1 autoimmune hepatitis were determined. METHODS: DNA samples from 155 patients and 102 ethnically similar normal individuals were assessed by polymerase chain reaction for polymorphisms of 4 different cytokine-producing genes. RESULTS: Only genotypes associated with the guanine to adenine substitution at position -308 of the tumor necrosis factor gene occurred more commonly in patients than in normal subjects (56% vs. 26%; P < 0.001). Patients with this polymorphism had the HLA DRB1*0301 allele (81% vs. 10%; P < 0.000001) and A1-B8-DRB1*0301 (66% vs. 0%; P < 0.000001) phenotype more frequently and HLA DRB1*04 alleles less often (24% vs. 67%; P < 0.000001). They also entered remission less commonly (56% vs. 78%; P = 0.01), had treatment failure more often (20% vs. 7%; P = 0.03), and developed cirrhosis more frequently (40% vs. 19%; P = 0.05). These latter differences, however, were not statistically significant by adjusted P value. CONCLUSIONS: A polymorphism of the tumor necrosis factor gene occurs more commonly in patients with type 1 autoimmune hepatitis than in normal subjects; it is associated with a poorer response to corticosteroids. The polymorphism may be inherited as part of the extended A1-B8-DRB1*0301 haplotype and may affect both disease expression and behavior.

CC-3052: a water-soluble analog of thalidomide and potent inhibitor of activation-induced TNF-alpha production.

The immunomodulatory drug thalidomide has been shown to be clinically useful in a number of situations due to its ability to inhibit TNF-alpha synthesis. However, its use is restricted by potentially serious side effects, including teratogenicity and neuorotoxicity; furthermore, insolubility may present problems in terms of systemic bioavailability. Recently, structural modifications of thalidomide have been designed enabling greatly enhanced anti-TNF-alpha activity in LPS-treated mice. In contrast to thalidomide (LPS-induced TNF-alpha IC50 approximately 200 microM in DMSO) and other analogs tested, one of these compounds, CC-3052 (IC50 approximately 1 microM in water), is water soluble. Furthermore, this analog exhibits increased stability in human plasma (t(1/2) approximately 17.5 vs 1.5 h for thalidomide) and appears to be nontoxic, nonmutagenic, and nonteratogenic. At pharmacologically active levels, cellular proliferation and LPS-induced IL-6 mRNA and IL-12p40 mRNA (as well as IL-1beta and IL-6 protein levels) in whole blood cultures were not affected; apparent inhibition of NK activity by CC-3052 was reversed upon addition of exogenous rTNF-alpha. In addition, IL-10 mRNA and protein levels were increased. These properties are consistent with results indicating inhibition of phosphodiesterase type IV activity by CC-3052. Furthermore, CC-3052 did not increase the degradation rate of macrophage TNF-alpha transcripts nor inhibit LPS-induced primary macrophage NF-kappaB activation. Taken together, the potency of selective TNF-alpha inhibition, water solubility, and increased plasma stability make CC-3052 an excellent candidate for further development and clinical evaluation for the treatment of TNF-alpha-mediated disease.

Tumour necrosis factor (TNF) and TNF-related molecules in HIV-1+ individuals: relationship with in vitro Th1/Th2-type response.

We examined the secretion and expression by peripheral blood mononuclear cells (PBMC) of TNF-alpha and TNF-related molecules with regard to Th1/Th2-type cytokine production. In 76 HIV+ patients at different disease stages and in 25 controls we measured cytokine (TNF-alpha/beta, interferon-gamma (IFN-gamma), IL-2, IL-4, IL-10), and activation marker secretion (sCD4, sCD8, sCD30) in phytohaemagglutinin (PHA)-stimulated and unstimulated PBMC cultures by ELISA, and membrane-bound TNF-alpha and CD30 expression by flow cytometry. We found an expansion of the TNF system in HIV+ individuals, that positively correlated with TNF-alpha, IFN-gamma and sCD8, probably representing activation of the cytotoxic compartment. In advanced disease these correlations disappeared, and TNF-alpha and TNF-related molecules positively correlated with IL-10. Our results are in line with the hypothesis that an expanded TNF system is immunopathological in conjunction with Th2-type immunity in the advanced stage of disease and with the inexorable progression to disease seen when both IL-10 and TNF-alpha are elevated.

Disco fever: epidemic meningococcal disease in northeastern Argentina associated with disco patronage.

Neisseria meningitidis is a leading cause of adult meningitis worldwide. From 5 to 14 August 1996, 8 cases of meningococcal disease occurred in Corrientes city (population 306,000) in northeastern Argentina. Those infected ranged in age from 15 to 45 years (median, 18.5). To determine risk factors for infection, a case-control study was done. Infecting isolates were serogrouped and underwent phenotyping by multilocus enzyme electrophoresis (MLEE) and pulsed-field gel electrophoresis (PFGE). Those infected were significantly more likely than those not infected to have had exposure to passive or active cigarette smoke or to have attended a particular disco. Isolates available from 6 case-patients were all serogroup C; all had identical MLEE and PFGE patterns. These data suggest that dance clubs or discos may be a focus of transmission of N. meningitidis among young people.

Abnormal intracellular IL-2 and interferon-gamma (IFN-gamma) production as HIV-1-assocated markers of immune dysfunction.

We used three-colour cytometry to analyse intracellular cytokine production in activated whole blood cultures derived from patients with HIV-1 infection. We assessed mitogen-induced IL-2, IL-4 and IFN-gamma production from T cells as possible markers of immune dysfunction. The percentages of T cells staining for IL-2 were significantly reduced in stimulated cultures from HIV+ individuals relative to normal controls (P<0.0001); this reduction was observed in both the CD4+ and the CD8+ subsets. IL-2 production was significantly reduced in CD4+ T cells from HIV+ individuals clinically classified as symptomatics compared with HIV+ asymptomatics (P<0.001); in addition, production of IL-2 inversely correlated with viral load (r2=0.832). On the other hand, HIV+ individuals showed significantly more T cells staining positive for IFN-gamma (P<0.0001); subset analysis identified these T cells as CD8+. Increased IFN-gamma production in the CD8+ T cell subset of HIV+ individuals correlated neither with clinical status nor with plasma viral load. IL-4 staining in activated T cells was low (<5%) and no differences were observed between HIV+ and control groups. Three-colour FACS analysis of whole blood provides a sensitive, rapid and relatively easy means to detect cytokine profiles within T cell subpopulations. Only small volumes of blood are required (0.5 ml), since there is no need for cell isolation, making it more practical than ELISA or reverse transcriptase-polymerase chain reaction (RT-PCR) for the analysis of immune function in HIV+ individuals. This technique could therefore play a role in mapping the dynamics and extent of immune recovery in AIDS patients undergoing triple combination therapy.

Liver failure and death after exposure to microcystins at a hemodialysis center in Brazil.

BACKGROUND: Hemodialysis is a common but potentially hazardous procedure. From February 17 to 20, 1996, 116 of 130 patients (89 percent) at a dialysis center (dialysis center A) in Caruaru, Brazil, had visual disturbances, nausea, and vomiting associated with hemodialysis. By March 24, 26 of the patients had died of acute liver failure. METHODS: A case patient was defined as any patient undergoing dialysis at dialysis center A or Caruaru's other dialysis center (dialysis center B) during February 1996 who had acute liver failure. To determine the risk factors for and the source of the outbreak, we conducted a cohort study of the 130 patients at dialysis center A and the 47 patients at dialysis center B, reviewed the centers' water supplies, and collected water, patients' serum, and postmortem liver tissue for microcystin assays. RESULTS: One hundred one patients (all at dialysis center A) met the case definition, and 50 died. Affected patients who died were older than those who survived (median age, 47 vs. 35 years, P<0.001). Furthermore, all 17 patients undergoing dialysis on the Tuesday-, Thursday-, and Saturday-night schedule became ill, and 13 of them (76 percent) died. Both centers received water from a nearby reservoir. However, the water supplied to dialysis center B was treated, filtered, and chlorinated, whereas the water supplied to dialysis center A was not. Microcystins produced by cyanobacteria were detected in water from the reservoir and from dialysis center A and in serum and liver tissue of case patients. CONCLUSIONS: Water used for hemodialysis can contain toxic materials, and its quality should therefore be carefully monitored.