Detection of Parechovirus (P) and Enterovirus (E) Among Infants Evaluated for Late-Onset Sepsis in the Neonatal Intensive Care Unit (NICU): The VIRIoN-P-E Study

Background: Limited data exist on the role of human parechoviruses (HPeV) and enteroviruses (EV) as causes of late-onset sepsis (LOS) in the NICU. Objective: To determine the frequency of detection of parechoviruses and enteroviruses among infants >72 hr of age who were evaluated for LOS in 2 academic NICUs (Parkland Memorial Hospital [PMH], Dallas -shared bays; Women & Infants Hospital [W&I], RI -single patient rooms) Design/Methods: Prospective cohort study of inborn infants hospitalized in the NICU at PMH and WIH from 1/2012 to 1/2013 and were enrolled in the Viral Respiratory Infections in the Neonatal Intensive Care Unit (VIRIoN-I; J Pediatr 2014:165:690). Eligible subjects were infants of all gestational ages (GA) and birth weights (BW) who were >72 hrs of age, remained in the NICU since birth, and underwent evaluation with initiation of antibiotic therapy for suspected LOS. Nasopharyngeal specimens were obtained within 72 hrs of the sepsis evaluation using flexible flocked nylon swabs that were placed in universal transport medium and frozen at -80\ub0C until tested for parechovirus and enterovirus RNA by polymerase chain reaction (PCR) assay (Virology Laboratory, Nationwide Children\u2019s Hospital, Columbus, OH). Demographic, clinical, laboratory, and radiographic data were obtained. Results: Of the 100 infants enrolled in the VIRIoN-I study, nasopharyngeal specimens were available from 65 (59, PMH; 6, WIH) for parechovirus and enterovirus PCR testing. These 65 infants (38, male; 27, female; 49, Hispanic; 6, white; 9, Black; 1, unknown) had a mean \ub1SD gestational age of 30 \ub1 5 wks and birth weight of 1619 \ub1 929 g, and received empirical antibiotics for possible LOS. Infants had a total of 94 sepsis evaluations (65, 1 evaluation; 16, 2; 8, 3; 4, 4) at a mean age of 20 days. Reasons for the sepsis evaluations included fever (9), hypothermia (65), apnea (50),feeding intolerance (51), seizure (1), irritabilitiy (5), emesis (20), diarrhea (1), bloody stool (5), rhinorrhea/congestion/cough (6), and lethargy (9). Four infants died. None of the infants had parechovirus or enterovirus detected in nasopharygeal specimens either at the first or subsequent sepsis evaluations. Conclusion(s): The burden of disease due to parechovirus and enteroviruses among inborn infants who remain in the NICU since birth appears to be low in those evaluated for LOS. Larger, prospective studies are needed to fully determine their contribution to \u201cculture-negative\u201d sepsis in the NICU. Publication Number: 3860.53

Meropenem vs standard of care for treatment of neonatal late onset sepsis (NeoMero1): A randomised controlled trial.

BACKGROUND: The early use of broad-spectrum antibiotics remains the cornerstone for the treatment of neonatal late onset sepsis (LOS). However, which antibiotics should be used is still debatable, as relevant studies were conducted more than 20 years ago, recruited in single centres or countries, evaluated antibiotics not in clinical use anymore and had variable inclusion/exclusion criteria and outcome measures. Moreover, antibiotic-resistant bacteria have become a major problem in many countries worldwide. We hypothesized that efficacy of meropenem as a broad-spectrum antibiotic is superior to standard of care regimens (SOC) in empiric treatment of LOS and aimed to compare meropenem to SOC in infants aged 44 weeks meeting the Goldstein criteria of sepsis, were randomized in a 1:1 ratio to receive meropenem or one of the two SOC regimens (ampicillin+gentamicin or cefotaxime+gentamicin) chosen by each site prior to the start of the study for 8-14 days. The primary outcome was treatment success (survival, no modification of allocated therapy, resolution/improvement of clinical and laboratory markers, no need of additional antibiotics and presumed/confirmed eradication of pathogens) at test-of-cure visit (TOC) in full analysis set. Stool samples were tested at baseline and Day 28 for meropenem-resistant Gram-negative organisms (CRGNO). The primary analysis was performed in all randomised patients and in patients with culture confirmed LOS. Proportions of participants with successful outcome were compared by using a logistic regression model adjusted for the stratification factors. From September 3, 2012 to November 30th 2014, total of 136 patients (instead of planned 275) in each arm were randomized; 140 (52%) were culture positive. Successful outcome at TOC was achieved in 44/136 (32%) in the meropenem arm vs. 31/135 (23%) in the SOC arm (p = 0.087). The respective numbers in patients with positive cultures were 17/63 (27%) vs. 10/77 (13%) (p = 0.022). The main reason of failure was modification of allocated therapy. Treatment emergent adverse events occurred in 72% and serious adverse events in 17% of patients, the Day 28 mortality was 6%. Cumulative acquisition of CRGNO by Day 28 occurred in 4% of patients in the meropenem and 12% in the SOC arm (p = 0.052). CONCLUSIONS: Within this study population, we found no evidence that meropenem was superior to SOC in terms of success at TOC, short term hearing disturbances, safety or mortality were similar in both treatment arms but the study was underpowered to detect the planned effect. Meropenem treatment did not select for colonization with CRGNOs. We suggest that meropenem as broad-spectrum antibiotic should be reserved for neonates who are more likely to have Gram-negative LOS, especially in NICUs where microorganisms producing extended spectrum- and AmpC type beta-lactamases are circulating

Identification and characterization of stimulator of interferon genes as a robust adjuvant target for early life immunization

Immunization is key to preventing infectious diseases, a leading cause of death early in life. However, due to age-specific immunity, vaccines often demonstrate reduced efficacy in newborns and young infants as compared to adults. Here, we combined in vitro and in vivo approaches to identify adjuvant candidates for early life immunization. We employed newborn and adult bone marrow-derived dendritic cells (BMDCs) to perform a screening of pattern recognition receptor agonists and found that the stimulator of interferon genes ligand 2'3'-cGAMP (hereafter cGAMP) induces a comparable expression of surface maturation markers in newborn and adult BMDCs. Then, we utilized the trivalent recombinant hemagglutinin (rHA) influenza vaccine, Flublok, as a model antigen to investigate the role of cGAMP in adult and early life immunization. cGAMP adjuvantation alone could increase rHA-specific antibody titers in adult but not newborn mice. Remarkably, as compared to alum or cGAMP alone, immunization with cGAMP formulated with alum (Alhydrogel) enhanced newborn rHA-specific IgG2a/c titers ~400-fold, an antibody subclass associated with the development of IFN\uce\ub3-driven type 1 immunity in vivo and endowed with higher effector functions, by 42 days of life. Highlighting the amenability for successful vaccine formulation and delivery, we next confirmed that cGAMP adsorbs onto alum in vitro. Accordingly, immunization early in life with (cGAMP+alum) promoted IFN\uce\ub3 production by CD4+T cells and increased the proportions and absolute numbers of CD4+CXCR5+PD-1+T follicular helper and germinal center (GC) GL-7+CD138+B cells, suggesting an enhancement of the GC reaction. Adjuvantation effects were apparently specific for IgG2a/c isotype switching without effect on antibody affinity maturation, as there was no effect on rHA-specific IgG avidity. Overall, our studies suggest that cGAMP when formulated with alum may represent an effective adjuvantation system to foster humoral and cellular aspects of type 1 immunity for early life immunization

Prevalence, Outcome, and Prevention of Congenital Cytomegalovirus Infection in Neonates Born to Women With Preconception Immunity (CHILd Study)

Background. Human cytomegalovirus (HCMV) is the leading infectious cause of congenital disabilities. We designed a prospective study to investigate the rate, outcome, and risk factors of congenital CMV (cCMV) infection in neonates born to immune women, and the potential need and effectiveness of hygiene recommendations in this population.Methods. The study was composed of 2 sequential parts: an epidemiology (part 1) and a prevention (part 2) study. Performance of part 2 depended upon a cCMV rate >0.4%. Women enrolled in part 1 did not receive hygiene recommendations. Newborns were screened by HCMV DNA testing in saliva and cCMV was confirmed by urine testing.Results. Saliva swabs were positive for HCMV DNA in 45/9661 newborns and cCMV was confirmed in 18 cases. The rate of cCMV was .19% (95% confidence interval [CI]: .11-.29%), and 3 out of 18 infants with cCMV had symptoms of CMV at birth. Age, nationality, occupation, and contact with children were similar between mothers of infected and noninfected newborns. Twin pregnancy (odds ratio [OR]: 7.2; 95% CI: 1.7-32.2; P=.037) and maternal medical conditions (OR: 3.9; 95% CI: 1.5-10.1; P= .003) appeared associated with cCMV. Given the rate of cCMV was lower than expected, the prevention part of the study was cancelled.Conclusions. Newborns from women with preconception immunity have a low rate of cCMV, which appears to be mostly due to reactivation of the latent virus. Therefore, serological screening in childbearing age would be pivotal to identify HCMV-seropositive women, whose newborns have a low risk of cCMV

Is Lactoferrin More Effective in Reducing Late-Onset Sepsis in Preterm Neonates Fed Formula Than in Those Receiving Mother's Own Milk? Secondary Analyses of Two Multicenter Randomized Controlled Trials

Background: Lactoferrin is the major antimicrobial protein in human milk. In our randomized controlled trial (RCT) of bovine lactoferrin (BLF) supplementation in preterm neonates, BLF reduced late-onset sepsis (LOS). Mother's own milk (MM) contains higher concentrations of lactoferrin than donor milk or formula, but whether BLF is more effective in infants who receive formula or donor milk is uncertain. Aim: To evaluate the incidence of LOS in preterm infants fed MM and in those fed formula and/or donor milk. Study Design: This is a (A) post hoc subgroup analysis, in our RCT of BLF, of its effects in preterm infants fed MM, with or without formula, versus those fed formula and/or donor milk (no-MM) and (B) post hoc meta-analysis, in our RCT of BLF and in the ELFIN (Enteral Lactoferrin in Neonates) RCT, of the effect of BLF in subgroups not exclusively fed MM. Results (A) Of 472 infants in our RCT, 168 were randomized to placebo and 304 were randomized to BLF. Among MM infants, LOS occurred in 22/133 (16.5%) infants randomized to placebo and in 14/250 (5.6%) randomized to BLF (relative risk or risk ratio (RR): 0.34; relative risk reduction (RRR): 0.66; 95% confidence interval (95% CI) for RR: 0.18-0.64; p < 0.0008). Among no-MM infants, LOS occurred in 7/35 (20.0%) randomized to placebo and in 2/54 (3.7%) randomized to BLF (RR: 0.19; RRR: 0.81; 95% CI for RR: 0.16-0.96; p = 0.026). In multivariable logistic regression analysis, there was no interaction between BLF treatment effect and type of feeding (p = 0.628). (B) In 1,891 infants not exclusively fed MM in our RCT of BLF and in the ELFIN RCT, BLF reduced the RR of LOS by 18% (RR: 0.82; 95% CI: 0.71-0.96; p = 0.01). Conclusion: Adequately powered studies should address the hypothesis that BLF is more effective in infants fed formula or donor milk than those fed MM. Such studies should evaluate whether a specific threshold of total lactoferrin intake can be identified to protect such patients from LOS

Producción de semillas de alfalfa (Medicago sativa) en el VBRC. Mitigación de la contaminación con transgenes

La contaminación de las semillas de alfalfa con variedades transgénicas o genéticamente modificadas (GM) preocupa a los semilleros instalados en el Valle Bonaerense del río Colorado (VBRC). En función de los últimos diagnósticos realizados, resultará muy difícil lograr que las semillas se encuentren completamente libres de impurezas GM. En este informe se describen las posibles fuentes de contaminación, para delinear estrategias de manejo a nivel de lote y de gestión a escala predial para evitar o minimizar la contaminación con transgenes. Se propone emplear un método de detección fenotípico/proteico específico desarrollado en el INTA Hilario Ascasubi para realizar controles de contaminación en diferentes instancias del entramado productivo comercial. El enfoque de organización por “barrios de producción” requiere una participación colaborativa de los actores de la cadena, pero a su vez presenta una oportunidad única para la valorización de la producción generada en la región. Se recomienda el establecimiento de umbrales de tolerancia de contaminantes GM para la producción de semilla de alfalfa convencional.EEA Hilario AscasubiFil: Renzi Pugni, Juan Pablo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Hilario Ascasubi; ArgentinaFil: Renzi Pugni, Juan Pablo. Universidad Nacional del Sur. Departamento de Agronomía; ArgentinaFil: Garcia, Flavia Carina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Hilario Ascasubi; ArgentinaFil: Crisanti, Paola Andrea. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Hilario Ascasubi; ArgentinaFil: Rodriguez, Graciela Adriana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Hilario Ascasubi; ArgentinaFil: Bruna, Matias Nicolas. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Hilario Ascasubi; ArgentinaFil: Quintana, Matias. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Hilario Ascasubi; ArgentinaFil: Reinoso, Omar Juan. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Hilario Ascasubi; ArgentinaFil: Vanzolini, Juan Ignacio. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Hilario Ascasubi; ArgentinaFil: Vanzolini, Juan Ignacio. Universidad Nacional del Sur. Departamento de Agronomía; ArgentinaFil: Coito, Carlos. Southern Seeds S.A.; ArgentinaFil: Ancia, V. Southern Seeds S.A.; ArgentinaFil: Guasch, P. Guasch Semillas; ArgentinaFil: Arditti, S. Guasch Semillas; ArgentinaFil: Gaido, E. Municipalidad de Patagones; ArgentinaFil: Cantamutto, Miguel Angel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Hilario Ascasubi; ArgentinaFil: Cantamutto, Miguel Angel. Universidad Nacional del Sur. Departamento de Agronomía; ArgentinaFil: Cantamutto, Miguel Angel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiarida; ArgentinaFil: Cantamutto, Miguel Angel. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiarida; Argentin

Prevalence, Outcome, and Prevention of Congenital Cytomegalovirus Infection in Neonates Born to Women with Preconception Immunity (CHILd Study)

Background: Human cytomegalovirus (HCMV) is the leading infectious cause of congenital disabilities. We designed a prospective study to investigate the rate, outcome, and risk factors of congenital CMV (cCMV) infection in neonates born to immune women, and the potential need and effectiveness of hygiene recommendations in this population. Methods: The study was composed of 2 sequential parts: an epidemiology (part 1) and a prevention (part 2) study. Performance of part 2 depended upon a cCMV rate >0.4%. Women enrolled in part 1 did not receive hygiene recommendations. Newborns were screened by HCMV DNA testing in saliva and cCMV was confirmed by urine testing. Results: Saliva swabs were positive for HCMV DNA in 45/9661 newborns and cCMV was confirmed in 18 cases. The rate of cCMV was. 19% (95% confidence interval [CI]:. 11-.29%), and 3 out of 18 infants with cCMV had symptoms of CMV at birth. Age, nationality, occupation, and contact with children were similar between mothers of infected and noninfected newborns. Twin pregnancy (odds ratio [OR]: 7.2; 95% CI: 1.7-32.2; P =. 037) and maternal medical conditions (OR: 3.9; 95% CI: 1.5-10.1; P =. 003) appeared associated with cCMV. Given the rate of cCMV was lower than expected, the prevention part of the study was cancelled. Conclusions: Newborns from women with preconception immunity have a low rate of cCMV, which appears to be mostly due to reactivation of the latent virus. Therefore, serological screening in childbearing age would be pivotal to identify HCMV-seropositive women, whose newborns have a low risk of cCMV. Clinical trials registration: www.clinicaltrials.gov (NCT03973359)

CMV DNA detection in dried samples of urine and saliva for congenital CMV diagnosis and screening

Background - Setting up neonatal screening for congenital CMV (cCMV) infection may be worthwhile for preventing the disability caused by the infection as the identified infected infants both symptomatic and asymptomatic at birth could be enrolled in a clinical and instrumental follow up to detect the damage early and start suitable interventions promptly. Detection of CMV DNA in neonatal dried blood spots (DBS test) has proven to be a screening method simpler faster and less costly than viral isolation. However the low viral load in the blood can negatively affect the results of DBS testing. Since urine or saliva of infected babies usually contain viral loads higher than those in the blood we tested for CMV DNA samples of urine dried on paper (DUS) and of saliva dried on nylon swabs (DSS) in the aim of identifying alternative methods for cCMV diagnosis/screening. Methods- Viral DNA was searched by means of nested PCR (gB gene) in mock DUS and DSS samples, spiked with 10-fold dilutions of cell grown CMV. The same test was employed on DUS and DSS collected from 337 unselected babies (2-48 days) and 26 cCMV infected infants (14 days-6 years) as controls. Conventional urine and saliva specimens from the same subjects were tested by means of n-PCR and shell-vial assays. DBS samples of positive cases were tested to assess the nature of the infection. Results \u2013 n-PCR detected CMV DNA down to 10E3 copies/ml in mock DUS and to 10E0 copies/ml in mock DSS. Both tests identified 7 infected infants, four with congenital and three with postnatal CMV infection. Fourteen further cases were positive only in the DSS test. Testing DUS and DSS showed 100% sensitivity and 95% specificity versus testing the corresponding liquid samples. The best performances of the tests were recorded in babies within their second life week. Conclusions \u2013 Detection of viral DNA in DUS and DSS seem to be reliable methods for diagnosing cCMV infection. The ease of collection of saliva could favor the use of DSS test in screening programs when the results obtained in this study will be confirmed