María Romero

Bacterial translocation in acute rejection after small bowel transplantation in rats

Acute rejection after small bowel transplantation (SBTx) may facilitate bacterial translocation (BT) and subsequent changes in the liver, spleen, and lungs. This study investigated whether BT occurs after acute rejection and whether this is followed by changes in the structure of the intestine and the phagocytic organs interposed between the gut and the general circulation. Orthotopic SBTx was performed in allogeneic (ALLO) rat-strain combinations (BN–Wistar, n=5). For comparison we used syngeneic SBTx (SYN) (BN–BN, n=6) controls. Animals were sacrificed on postoperative day 7. Mesenteric lymph nodes and portal and caval blood were cultured for aerobes and anaerobes. Escherichia coli β-galactosidase DNA was assessed by polymerase chain reaction in the blood samples. Intestine, liver, spleen, and lung protein and DNA contents were measured. Histologic changes were graded according to standard criteria of acute rejection. For comparisons we used chi2 and nonparametric Mann–Whitney test with a threshold of significance of p<0.05. ALLO rats lost more weight after SBTx than SYN rats (−13.02±4.39% vs. −8.04±5.08% of preoperative weight), although the difference was not significant (ns). A variable degree of graft rejection was histologically demonstrated in all ALLO rats, and DNA/protein content in the graft was significantly higher in this group (0.245±0.85 vs. 0.134±0.21, p<0.05). Gram-negative enteric bacteria were found in 4/5 ALLO and 4/6 SYN rats (ns), and aerobic Gram-positive bacteria in 2/5 and 3/6 (ns), respectively. Anaerobic growth occurred in mesenteric lymph nodes in one ALLO rat and in the bloodstream in another one. E. coli DNA was isolated in none of the ALLO but in two SYN rats (ns). BT was frequent after SBTx in both syngeneic and allogeneic strain combinations. Contrary to our expectations, BT after SBTx was not higher in ALLO group rats. However, anaerobic germs were isolated only in this group.

Human papillomavirus mRNA testing for the detection of anal high‐grade squamous intraepithelial lesions in men who have sex with men infected with HIV

Currently, screening for anal high‐grade squamous intraepithelial lesions (anal HSIL) relies on anal cytology and high‐resolution anoscopy. Since this approach has limited sensitivity and specificity for detecting anal HSIL, there is increasing interest in the role of biomarkers for predicting anal HSIL. The aim of this study is to evaluate the diagnostic accuracy of HPV E6/E7‐mRNA expression for the detection of anal HSIL in MSM infected with HIV, in comparison to DNA‐HR‐HPV and anal cytology. This cross‐sectional screening study included 101 MSM followed at the HIV‐unit of La Paz University Hospital. Intra‐anal swabs from patients participating in a screening program including cytology, high‐resolution anoscopy and histology were analyzed. HR‐HPV‐DNA detection was performed by means of the CLART® HPV2 assay (GENOMICA S.A.U., Madrid, Spain). E6/E7‐mRNA detection of HR‐HPV‐types 16, 18, 31, 33, and 45 was performed using the NucliSENS‐EasyQ assay (BioMérieux, Marcy ĺEtoile, France). HR‐HPV DNA and HPVE6/E7 mRNA were detected in 82% and 57% of the anal smears respectively. Anal cytology screening was abnormal in 70.3%. For the detection of HSIL sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were 71.7%, 55.6%, 57.9%, and 69.8% for E6/E7‐mRNA testing, respectively, compared to 100%, 31.5%, 55.4%, and 100% for HR‐HPV‐DNA testing and to 83%, 40.7%, 54.9%, 73.3% of cytology testing. In comparison with the other tests, HPVE6/E7 mRNA testing yielded a lower clinical sensitivity but a higher clinical specificity and PPV for the detection of anal HSIL in MSM infected with HIV. J. Med. Virol. 87:1397–1403, 2015.

Utilidad de la terapia intracoronaria con células progenitoras en pacientes con miocardiopatía dilatada: ¿puente o alternativa al trasplante cardiaco?

INTRODUCTION AND OBJECTIVES Some paediatric publications have recently raised the value of intracoronary therapy with autologous bone marrow-derived progenitor cells (APCs) in children with dilated cardiomyopathy (DCM) and heart failure. We describe the usefulness of this treatment in two infants with severe DCM and heart failure, who had been transferred to our hospital for cardiac transplant evaluation. PATIENTS AND METHODS The first patient was a 3 months old male weighing 4 kg. The second was a 4 months old male weighing 5 kg. At the time of admission, both were in poor clinical condition (NYHA IV), with severe dilation and systolic dysfunction (ejection fraction [EF]<30%) of the left ventricle and marked elevation of NT-proBNP, requiring treatment with mechanical ventilation and inotropic iv infusion. After mobilization with G-CSF for 4 days, APCs were obtained from peripheral blood by leukocytapheresis, administering them by a slow intracoronary bolus injection using a stop-flow technique (6.15x106 CD34-positive cells/Kg in the first patient, and 10.55x106 CD34-positive cells/Kg in the second). RESULTS Since the first week after the procedure, clinical status of patients improved and echocardiography showed a decrease in left ventricular dilation. A month later, there was a significant improvement in EF (> 40%) and NT-proBNP levels, subsequently maintained throughout the follow-up. However, four months later in the first patient, the left ventricle dilated again and its function slightly worsened, but without any significant impact in his clinical status. CONCLUSIONS Intracoronary therapy with APCs can be an alternative in children, especially infants, with DCM and heart failure. It can reduce the waiting list mortality, improve clinical status and provide more time on the waiting list to receive a suitable organ, or even to make transplantation unnecessary.

High MICs for Vancomycin and Daptomycin and Complicated Catheter-Related Bloodstream Infections with Methicillin-Sensitive Staphylococcus aureus

Patients infected with these bacteria were more likely to have local endovascular complications.

Molecular epidemiology of enterovirus and parechovirus infections according to patient age over a 4‐year period in Spain

The epidemiology and clinical association of enterovirus (EV) and parechovirus (HPeV) infections, as well as the type‐distribution‐according‐to‐age, were determined during a 4‐year study period in Spain. During 2010–2013, a total of 21,832 clinical samples were screened for EV and the detection frequency was 6.5% (1,430). Of the total EV‐negative samples, only 1,873 samples from 2011 to 2013 were available for HPeV testing. HPeV was detected in 42 (2%) of them. Positive samples were genotyped using PCR and sequencing. EV infections occurred in all age groups of patients: neonates (17%), children 28 days to 2 years (29%), children 2–14 years (40%), and adults (14%). Thirty‐four different EV types were identified. HPeV infections were detected exclusively in infants <8 m (70% neonates, P < 0.05). All but one HPeV were HPeV‐3. Differences in type frequency detection were found according to age and clinical manifestation. Coxsackievirus (CV)‐B4 (61%), CV‐B5 (83%), and HPeV‐3 (64%) were more frequent in neonates than in older patients (P < 0.05). Echovirus (E)‐3 (60%), E‐18 (47%), E‐25 (62%), CV‐A6 (61%), CV‐A16 (72%), and EV‐71 (75%) were mainly detected in children 28 days to 2 years (P < 0.05), whereas, E‐6 (79%), E‐20 (88%), and E‐30 (85%) were predominant in children >2 years and adults (P < 0.05). Clinically, meningitis was associated with EV (P < 0.01) whereas, encephalitis was more frequent in HPeV‐infected patients. CV‐B types were associated with myocarditis (90%; P < 0.05) and EV species A with hand–foot–mouth‐disease/atypical exanthema (88%; P < 0.05). J. Med. Virol. 89:435–442, 2017.

Pathogen-related factors affecting outcome of catheter-related bacteremia due to methicillin-susceptible Staphylococcus aureus in a Spanish multicenter study

Even with appropriate clinical management, complicated methicillin-susceptible Staphylococcus aureus (MSSA) catheter-related bacteremia (CRB) is frequent. We investigated the influence of molecular characteristics of MSSA strains on the risk of complicated bacteremia (CB) in MSSA-CRB. A multicenter prospective study was conducted in Spain between 2011 and 2014 on MSSA-CRB. Optimized protocol-guided clinical management was required. CB included endocarditis, septic thrombophlebitis, persistent bacteremia and/or end-organ hematogenous spread. Molecular typing, agr functionality and DNA microarray analysis of virulence factors were performed in all MSSA isolates. Out of 83 MSSA-CRB episodes included, 26 (31.3%) developed CB. MSSA isolates belonged to 16 clonal complexes (CCs), with CC30 (32.5%), CC5 (15.7%) and CC45 (13.3) being the most common. Comparison between MSSA isolates in episodes with or without CB revealed no differences regarding agr type and functionality. However, our results showed that CC15 and the presence of genes like cna, chp and cap8 were associated with the development of CB. The multivariate analysis highlighted that the presence of cna (Hazard ratio 2.9; 95% CI 1.14–7.6) was associated with the development of CB. Our results suggest that particular CCs and specific genes may influence the outcome of MSSA-CRB.

Enterovirus neurological disease and bacterial coinfection in very young infants with fever

BACKGROUND Very little information exists on simultaneous infections by viruses and bacteria in infants with fever without source (FWS). OBJECTIVES To investigate the incidence of bacterial coinfection in infants up to 3 months of age with neurological viral infection. STUDY DESIGN Prospective study performed in infants below 90 days of age attending the emergency room of two public hospitals in Spain for FWS. Those who had viral screening performed in CSF, together with blood, CSF and urine cultures were included. Herpes virus, EV and HPeV detection in CSF was performed by PCR. Coinfections between viruses in CSF and serious bacterial infections were described. RESULTS 119 Infants less than 90 days of age were recruited. Forty-five (38%) had viral infection of the central nervous system, and in 8 of them (17.7%) we found a concurrent bacterial infection: 7 urinary tract infections (UTI) and 1 sepsis. In all cases, the virus identified in CSF was EV. CONCLUSIONS Bacterial infections were frequent in young infants with viral neurological infections associated to EV. Urinary tract infection was the most common bacterial disease.

Pathological mechanisms of left main stent failure

BACKGROUND Despite the increasing use of left main (LM) percutaneous coronary intervention (LM-PCI), there have been no pathological studies devoted to understanding the causes of LM stent failure. We aimed to systematically determine the pathological mechanisms of LM stent failure. METHODS AND RESULTS From the CVPath Stent registry, a total of 46 lesions were identified to have LM-PCI. Pathologic stent failure (PSF) was defined as stent thrombosis, restenosis and in-stent chronic total occlusion (CTO). Failed and patent LM stented lesions were pathologically assessed to determine predictors of PSF. Malapposition and uncovered struts were numerically greater in the LM ostium, body, and bifurcation while neointimal thickness was relatively greater in bifurcation and proximal circumflex. In this study cohort, half of the lesions (n = 23) showed PSF. Stent thrombosis (ST, n = 18) was the major mode of PSF followed by in-stent CTO (n = 4) and restenosis (n = 1). Failed lesions showed significantly greater prevalence of malapposition >20% of struts/section (65% vs. 13%, P < 0.01), stent struts crossing an ostial side branch >30% of the circumference (48% vs. 13%, P < 0.01) and uncovered struts >30% (57% vs. 18%, P = 0.03). In multivariate analysis, the prevalence of malapposition >20% was the strongest risk factor for PSF (Odds ratio 8.0, 95% confidence interval 1.8-45.4, P < 0.01) followed by struts crossing an ostial side branch >30% (Odds ratio 4.2, 95% confidence interval 0.8-24.7, P = 0.09). CONCLUSION Our data demonstrate the main pathological predictors for LM stent failure are malapposition and struts crossing an ostial side branch and suggest that imaging-guided PCI may be important.

Difficulties in establishing the source of infection in recurrent neonatal group B streptococcal disease

Recurrent episodes of GBS invasive infections occurred 2, 10, and 50 days after completion of antibiotic therapy. The premature infant recurred 2 days after completion antibiotic therapy and the infant with early onset disease 50 days after completion antibiotic therapy. All patients had sepsis, one of them with meningitis. At the time of recurrence, rectovaginal and throat swabs cultures were positive for GBS in one mother with negative antenatal screening and in her infant’s throat swab culture. Mother’s rectovaginal culture and infant’s and parent’s throat swab cultures from the other two cases were negative, including the preterm case. Breast milk cultures were sterile in all cases. Complete physical examination, cerebral, cardiac and hip ultrasounds, and bone scan did not reveal occult bacterial foci in any case. Penicillin tolerance in GBS strains (from both the first episode and the recurrence) were studied in two cases, observing very low minimal inhibitory and bactericidal concentration (≤0, 25 mcg/ml for penicillin). The three infants were treated with intravenous ampicillin and recovered uneventfully without further relapses. Pathogenesis of recurrent GBS disease is poorly understood [2], and in many infants, the source of reinfection cannot be determined [2]. Matsubara et al. discussed in their article how prematurity could be a risk for recurrence and they suggested that human milk could serve as a source of repeated infant exposures [1, 3]. Prematurity has been reported as an important risk factor for recurrent GBS infection [2, 4]. The use of broad-spectrum antibiotics in GBS colonized premature newborns and the presence of reduced baseline levels of immunoglobulins may likely contribute to intestinal translocation [4]. In Matsubara et al. report, infants with late-onset disease were more likely to be premature [1]. However, analyzing the article data, there are no statistically significant differences between the prematurity rate observed in all GBS infections (27%; We have read with interest the article by Matsubara et al. describing the epidemiology of group B Streptococcus (GBS) disease in infants in Japan [1]. The authors reported 12 GBS recurrent infections, which is the largest series ever reported. We have recently performed a retrospective review of medical records of infants less than 3 months with GBS invasive disease who developed a recurrent infection during a 15-year period (2000–2014). During the study period, 117 GBS neonatal infections were diagnosed in our hospital (53 early onset and 64 late-onset diseases), three of which relapsed. This represents an incidence of recurrent infection of 2.6%, which is similar to that reported by Matsubara et al. and by other authors [1, 2]. Infants with recurrent invasive GBS infection had been born by vaginal delivery without prolonged rupture of membranes. Two of them were full term and one was preterm (born at 31 weeks of gestation). Antenatal rectovaginal cultures were negative in two cases and were not performed in the premature newborn. All infants were breastfed, with no history of mastitis in their mothers. First episodes of GBS invasive infections occurred as late-onset sepsis without meningitis in two infants (at the age of 12 and 23 days) and early onset sepsis in one (during the first day of life). The premature infant developed the disease at the age of 23 days. All patients were treated with intravenous ampicillin (200 mg/kg) for 10 days.

Características epidemiológicas y clínicas de los lactantes hospitalizados por infecciones por parechovirus humanos. Estudio prospectivo en España

INTRODUCTION Human parechovirus (HPeV) is one of the recently described picornaviridae viruses that have been associated with fever of unknown origin (FUO), clinical sepsis, gastroenteritis, meningitis, or encephalitis in very young infants. The aim of this study is to describe the epidemiology and clinical features of these viruses. PATIENTS AND METHODS A prospective multicentre 3-year study was conducted in 12 hospitals in Spain. Out of 850 specimens examined, 47 were positive (5.52%), with HPeV-3 being the most frequent (29 cases). Infections occurred throughout the year, but mainly in May and July, and a biennial distribution was observed. More than half (57%) were neonates, and only 2 children were older than 3 months. Fever was present in all children, with irritability in 45%, rash in 18.6%, and diarrhoea in 14%. The results of biochemical tests were all in normal range. The most common final diagnosis was FUO (61%), followed by clinical sepsis (29%). Up to 29% of infants were admitted to the intensive care unit, but only one patient had sequelae. RESULTS Out of 850 specimens examined, 47 were positive (5.52%) for HPeV, with HPeV-3 being the most frequent (29 cases). Infections occurred throughout the year, but mainly in May and July, and a biennial distribution was observed. More than half (57%) were neonates, and only 2 children were older than 3 months. Fever was present in all children, with irritability in 45%, rash in 18.6%, and diarrhoea in 14%. The results of biochemical tests were all in normal range. The most common final diagnosis was FUO (61%), followed by clinical sepsis (29%). Up to 29% of infants were admitted to the intensive care unit, but only one patient had sequelae CONCLUSIONS: HPeV circulates in our country, mainly during spring and summer, and affects young infants with a FUO and clinical sepsis. Molecular diagnostic techniques in all hospitals could help in improving the management of patients with these infections.