Benefits of a Pediatric Antimicrobial Stewardship Program in Antimicrobial Use and Quality of Prescriptions in a Referral Children's Hospital.

OBJECTIVES: To evaluate the results of the first 24 months of a postprescription review with feedback-based antimicrobial stewardship program in a European referral children's hospital. STUDY DESIGN: We performed a pre-post study comparing antimicrobial use between the control (2015-2016) and the intervention periods (2017-2018) expressed in days of therapy/100 days present. Quality of prescriptions was evaluated by quarterly cross-sectional point-prevalence surveys. Length of stay, readmission rates, in-hospital mortality rates, cost of systemic antimicrobial agents, and antimicrobial resistance rates were included as complementary outcomes. RESULTS: Total antimicrobial use and antibacterial use significantly decreased during the intervention period (P = .002 and P = .001 respectively), and total antifungal use remained stable. A significant decline in parenteral antimicrobial use was also observed (P < .001). In 8 quarterly point-prevalence surveys (938 prescriptions evaluated), the mean prevalence of use of any antimicrobial among inpatients was 39%. An increasing trend in the rate of optimal prescriptions was observed after the first point-prevalence survey (P = .0898). Nonoptimal prescriptions were more common in surgical than in medical departments, in antibacterial prescriptions with prophylactic intention, and in empirical more than in targeted treatments. No significant differences were observed in terms of mortality or readmission rates. Only minor changes in antimicrobial resistance rates were noted. CONCLUSIONS: Our antimicrobial stewardship program safely decreased antimicrobial use and expenditure, and a trend toward improvement in quality of prescription was also observed.

Diversity of Y-chromosomal and mtDNA Markers Included in Mediscope Chip within Two Albanian Subpopulations from Croatia and Kosovo: Preliminary Data.

The aim of this preliminary study is to analyze genetic specificity of Kosovo Albanians comparing with neighboring populations using new genetic tool - MEDISCOPE gene chip, to investigate the feasibility of this approach. We collected 37 DNA samples (9 Croats, 17 Albanians from Croatia and 11 Albanians from Kosovo) from unrelated males born in Croatia and Kosovo. Additionally, samples were expanded with female individuals and mtDNA analysis included a total of 61 samples (15 Croats, 23 Albanians from Croatia and 23 Albanians from Kosovo). This pilot study suggests that the usage of the MEDISCOPE chip could be recognized as an efficient tool within recognition of the population genetic specificity even within extremely small sample size.

Factors affecting levels of urinary albumin excretion in the general population of Spain: the Di@bet.es study.

The present study was undertaken to examine the prevalence of urinary ACR (albumin/creatinine ratio) >30 mg/g and the associated clinical and environmental factors in a representative sample of the population of Spain. Di@bet.es study is a national, cross-sectional population-based survey conducted in 2009-2010. Clinical, metabolic, socio-demographic, anthropometric data and information about lifestyle habit were collected. Those subjects without KDM (known diabetes mellitus) were given an OGTT (oral glucose tolerance test). Albumin and creatinine were measured in a urinary sample and ACR was calculated. The population prevalence of ACR >30 mg/g was 7.65% (adjusted for sex and age). The prevalence of ACR >30 mg/g increased with age (P<0.001). Subjects with carbohydrate metabolism disorders had a greater prevalence of ACR >30 mg/g but after being adjusted for age, sex and hypertension, was significant only in those subjects with UKDM (unknown diabetes mellitus) {OR (odd ratio), 2.07 [95% CI (confidence interval), 1.38-3.09]; P<0.001] and KDM [OR, 3.55 (95% CI, 2.63-4.80); P<0.001]. Prevalence of ACR >30 mg/g was associated with hypertension [OR, 1.48 (95% CI, 1.12-1.95); P=0.001], HOMA-IR (homoeostasis model assessment of insulin resistance) [OR, 1.47 (95% CI, 1.13-1.92); P≤0.01], metabolic syndrome [OR, 2.17 (95% CI, 1.72-2.72); P<0.001], smoking [OR, 1.40 (95% CI, 1.06-1.83); P≤0.05], physical activity [OR, 0.68 (95% CI, 0.54-0.88); P≤0.01] and consumption of fish [OR, 0.38 (95% CI, 0.18-0.78); P≤0.01]. This is the first study that reports the prevalence of ACR >30 mg/g in the Spanish population. The association between clinical variables and other potentially modifiable environmental variables contribute jointly, and sometimes interactively, to the explanation of prevalence of ACR >30 mg/g. Many of these risk factors are susceptible to intervention.

Assessing the Impact of Different Technological Strategies on the Fate of Salmonella in Chicken Dry-Fermented Sausages by Means of Challenge Testing and Predictive Models.

Salmonella is the main relevant pathogen in chicken dry-fermented sausages (DFS). The safety of shelf-stable DFS must rely on the production process, which should not only prevent growth but promote inactivation of Salmonella. The aim of the study was to assess the behaviour of Salmonella during the production process of two types of low-acid chicken DFS. The impact of the use of starter culture, corrective storage and high-pressure processing (HPP) at different processing times was assessed through challenge testing, i.e., inoculating a cocktail of Salmonella into the meat batter (at 6 Log10 cfu/g) used for sausage manufacture. Sausages of medium (fuet-type, FT) and small (snack-type, ST) calibre were elaborated through ripening (10-15 °C/16 d) and fermentation plus ripening (22 °C/3 d + 14 °C/7 d). Physico-chemical parameters were analysed and Salmonella was enumerated throughout the study. The observed results were compared with the simulations provided by predictive models available in the literature. In FT, a slight decrease in Salmonella was observed during the production process while in ST, a 0.9-1.4 Log10 increase occurred during the fermentation at 22 °C. Accordingly, DFS safety has to be based on the process temperature and water activity decrease, these factors can be used as inputs of predictive models based on the gamma-concept, as useful decision support tool for producers. Salmonella lethality was enhanced by combining HPP and corrective storage strategies, achieving >1 and 4 Log10 reductions for FT and ST, respectively.

Growth and Non-Thermal Inactivation of Staphylococcus aureus in Sliced Dry-Cured Ham in Relation to Water Activity, Packaging Type and Storage Temperature.

Dry-cured ham (DCH) could support the growth of Staphylococcus aureus as a halotolerant bacterium, which may compromise the shelf-stability of the product according to the growth/no growth boundary models and the physicochemical parameters of commercial DCH. In the present study, the behavior of S. aureus is evaluated in sliced DCH with different water activity (aw 0.861-0.925), packaged under air, vacuum, or modified atmosphere (MAP), and stored at different temperatures (2-25 °C) for up to 1 year. The Logistic and the Weibull models were fitted to data to estimate the primary kinetic parameters for the pathogen Log10 increase and Log10 reduction, respectively. Then, polynomial models were developed as secondary models following their integration into the primary Weibull model to obtain a global model for each packaging. Growth was observed for samples with the highest aw stored at 20 and 25 °C in air-packaged DCH. For lower aw, progressive inactivation of S. aureus was observed, being faster at the lowest temperature (15 °C) for air-packaged DCH. In contrast, for vacuum and MAP-packaged DCH, a higher storage temperature resulted in faster inactivation without a significant effect of the product aw. The results of this study clearly indicate that the behavior of S. aureus is highly dependent on factors such as storage temperature, packaging conditions and product aw. The developed models provide a management tool for evaluating the risk associated with DCH and for preventing the development of S. aureus by selecting the most appropriate packaging according to aw range and storage temperature.

Control of Listeria monocytogenes in chicken dry-fermented sausages with bioprotective starter culture and high-pressure processing.

Listeria monocytogenes is one of the most relevant pathogens for ready-to-eat food, being a challenge for the food industry to comply with microbiological criteria. The aim of the work was to assess the behavior of L. monocytogenes in two types of chicken-based dry-fermented sausages during the fermentation and ripening, with or without a bioprotective starter culture (Latilactobacillus sakei CTC494). To complement the challenge testing approach, simulations with different predictive models were performed to better understand the role of contributing factors. The impact of post-processing strategies, such as high-pressure processing and/or corrective storage was assessed. The chicken meat was inoculated with a cocktail of three L. monocytogenes strains, mixed with other ingredients/additives and stuffed into small (snack-type) or medium (fuet-type) casings. Snack-type was fermented (22°C/3 days) and ripened (14°C/7 days), while fuet-type was ripened (13°C/16 days). At the end of ripening, HPP (600 MPa/5 min) and/or corrective storage (4 or 15°C/7 days) were applied. The suitability of HPP after fermentation was evaluated in the snack-type sausages. Pathogen growth (>3 Log10) was observed only during the fermentation of the snack type without a starter. The bioprotective starter prevented the growth of L. monocytogenes in the snack-type sausages and enhanced the inactivation (1.55 Log10) in fuet-type sausages, which could be related to the higher lactic acid production and consequent decrease of pH, but also the production of the antilisterial bacteriocin sakacin k. The gamma concept model allowed us to identify the main factors controlling the L. monocytogenes' growth, i.e., the temperature during the early stages and a w at the end of the production process. The earlier acidification linked with the addition of starter culture made the interaction with the other factors (undissociated lactic acid, a w and temperature) to be the growth-preventing determinants. High-pressure processing only caused a significant reduction of L. monocytogenes in snack-type, which showed higher a w . The application of HPP after fermentation did not offer a relevant advantage in terms of efficacy. Corrective storage did not promote further pathogen inactivation. The findings of the work will guide the food industry to apply effective strategies (e.g., fermentation temperature and bioprotective starter cultures) to control L. monocytogenes in chicken dry-fermented sausages.

Closed-Loop Control of Postprandial Glycemia Using an Insulin-on-Board Limitation Through Continuous Action on Glucose Target.

BACKGROUND: Postprandial (PP) control remains a challenge for closed-loop (CL) systems. Few studies with inconsistent results have systematically investigated the PP period. OBJECTIVE: To compare a new CL algorithm with current pump therapy (open loop [OL]) in the PP glucose control in type 1 diabetes (T1D) subjects. METHODS: A crossover randomized study was performed in two centers. Twenty T1D subjects (F/M 13/7, age 40.7 ± 10.4 years, disease duration 22.6 ± 9.9 years, and A1c 7.8% ± 0.7%) underwent an 8-h mixed meal test on four occasions. In two (CL1/CL2), after meal announcement, a bolus was given followed by an algorithm-driven basal infusion based on continuous glucose monitoring (CGM). Alternatively, in OL1/OL2 conventional pump therapy was used. Main outcome measures were as follows: glucose variability, estimated with the coefficient of variation (CV) of the area under the curve (AUC) of plasma glucose (PG) and CGM values, and from the analysis of the glucose time series; mean, maximum (Cmax), and time to Cmax glucose concentrations and time in range (<70, 70-180, >180 mg/dL). RESULTS: CVs of the glucose AUCs were low and similar in all studies (around 10%). However, CL achieved greater reproducibility and better PG control in the PP period: CL1 = CL2 0.05) nor the need for oral glucose was significantly different (CL 40.0% vs. OL 22.5% of meals; P = 0.054). CONCLUSIONS: This novel CL algorithm effectively and consistently controls PP glucose excursions without increasing hypoglycemia. Study registered at ClinicalTrials.gov : study number NCT02100488.

Overexpression of FABP7 in Down syndrome fetal brains is associated with PKNOX1 gene-dosage imbalance.

Suppression subtractive hybridization performed on Down syndrome (DS) fetal brains revealed a differentially expressed gene, FABP7, mapped to 6q22-23. FABP7 overexpression in DS brains was verified by real-time PCR (1.63-fold). To elucidate the molecular basis of FABP7 overexpression and establish the relationship with chromosome 21 trisomy, the FABP7 promoter was cloned by genomic inverse PCR. Comparison to the mouse ortholog revealed conservation of reported regulatory elements, among them a Pbx/POU binding site, known to be the target of PBX heteromeric complexes. PBX partners include homeobox-containing proteins, such as PKNOX1 (PREP1), a transcription factor mapping at 21q22.3. We report here: (i) overexpression of PKNOX1 in DS fetal brains; (ii) in vitro specific binding of PKNOX1 to the Pbx/POU site of the FABP7 promoter; (iii) in vivo FABP7 promoter trans-activation in cultured neuroblastoma cells caused by PKNOX1 overexpression. To our knowledge this is the first report of a direct relation between dosage imbalance of a chromosome 21 gene and altered expression of a downstream gene mapping on another chromosome. Given the role of FABP7 in the establishment, development and maintenance of the CNS, we suggest that the overexpression of FABP7 could contribute to DS-associated neurological disorders.

Low Physical Activity and Its Association with Diabetes and Other Cardiovascular Risk Factors: A Nationwide, Population-Based Study.

Low physical activity (PA), or sedentary lifestyle, is associated with the development of several chronic diseases. We aimed to investigate current prevalence of sedentariness and its association with diabetes and other cardiovascular risk factors. PA was evaluated in a population-based, cross-sectional, randomly sampled study conducted in 2009-2010 in Spain. International Physical Activity Questionnaire (SF-IPAQ) was used to assess PA. 4991 individuals (median age 50 years, 57% women) were studied. Prevalence of sedentariness was 32.3% for men and 39% for women (p < 0.0001). Sex differences were particularly notable (age*sex interaction, p = 0.0024) at early and older ages. Sedentary individuals had higher BMI (28 vs. 27 kg/m2) and obesity prevalence (37 vs. 26%). Low PA was present in 44, 43, and 38% of individuals with known diabetes (KDM), prediabetes/unknown-diabetes (PREDM/UKDM), and normal glucose regulation (p = 0.0014), respectively. No difference between KDM and PREDM/UKDM (p = 0.72) was found. Variables independently associated (p < 0.05) with sedentariness were age, sex, BMI, central obesity, Mediterranean diet adherence, smoking habit, HDL-cholesterol, triglycerides and dyslipidemia. Low PA is on the rise in Spain, especially among women. Sedentariness is associated with several cardiovascular risk factors and may be responsible for the increasing prevalence of obesity and diabetes in this country.

Haptoglobin genotype and risk of diabetic nephropathy in patients with type 1 diabetes mellitus: a study on a Spanish population.

BACKGROUND: Few reports have studied the possible association between the haptoglobin (Hp) genotype and the risk of diabetic nephropathy (DN) in type 1 diabetes (T1D), with conflicting results to date. AIMS: To study whether the 2-2 Hp genotype is associated with an increased risk of overt DN in a Spanish population with T1D. METHODS: We performed a case-control study in a Spanish population. CASES: T1D patients with end-stage renal disease (stage 5 of NKF-KDOQI), awaiting reno-pancreatic transplantation or having already been transplanted (reno-pancreatic or renal alone). CONTROLS: T1D patients, matched for sex and time of diabetes evolution, with preserved renal function and normal urinary albumin excretion. Hp genotyping was done using polymerase chain reaction and electrophoresis. RESULTS: We included 57 cases and 57 controls in the study. There were no statistically significant differences in gender (70% vs. 61% males, p=1.0) or the duration of diabetes (23.0 ± 6.7 vs. 20.8 ± 9.3 years; p=0.1), although the age of onset of diabetes was lower in the cases (14.1 ± 6.8 vs. 17.7 ± 10.1 years, p=0.03). The frequency of genotypes 1-1, 1-2 and 2-2 was 19.3%, 42.1% and 38.6% in cases and 17.5%, 49.1% and 33.4% in controls, respectively, with no statistically significant differences between groups (p=0.8). Conditional logistic regression analysis showed no significant association between genotype 2-2 of Hp and the development of DN (OR 1.14, CI 0.52-2.52). CONCLUSIONS: In our sample of a Spanish population with T1D, no association was found between the Hp genotype and risk of overt DN.

Prevalence of the metabolic syndrome in Spain using regional cutoff points for waist circumference: the di@bet.es study.

The aim of the study is to assess the prevalence of metabolic syndrome (MetS) in Spain using specific cutoff points for waist circumference (WC) (>94.5 cm for men and >89.5 cm for women) and evaluating the influence of several socio-demographic and economic factors. Data on MetS were obtained from a national study of 4,727 subjects from 18 to 90 years of age, conducted in Spain between 2009 and 2010 (The di@bet.es study). MetS was defined applying the new Harmonized definition (evaluating the use of abdominal obesity (AO) as a obligatory criterion for MetS or not) as well as with other widely used criteria. Results were then compared with data from previous studies. Multiple logistic regression models were used to evaluate the influence of different social factors. The age-standardized MetS prevalence was 38.37 % (CI 35.74-40.99) in men and 29.62 % (CI 27.56-31.69) in women, when AO was required as a diagnostic criterion; 42.13 % (CI 39.37-44.89) and 32.31 % (CI 30.15-34.47) in men and women, respectively, if AO was not considered mandatory. Prevalence of MetS increased with age (p < 0.001 for trend). Women with a lower educational level were more likely to have MetS (OR 4.4; 95 % CI: 2.84-6.7) as compared with those with a higher educational level. Subjects with MetS had a worse physical quality of life. The combination of AO, hypertension and carbohydrate alterations was the most common MetS' pattern. A high prevalence of MetS was detected in the Spanish population especially in men, the elderly and women with a low educational level.

Use of drugs related to the treatment of diabetes mellitus and other cardiovascular risk factors in the Spanish population. The Di@bet.es study.

INTRODUCTION AND OBJECTIVES: To assess the patterns of use of 8 therapeutic drug groups for the treatment of diabetes mellitus and other cardiovascular risk factors, and to identify sociodemographic and health determinants of their use in the overall Spanish population. METHODS: A representative sample of the Spanish population within the Di@bet.es study, a cross-sectional population-based survey, was included. STUDY VARIABLES: sociodemographic, clinical, and lifestyle data; physical examination, and an oral glucose tolerance test in patients without known diabetes mellitus. Furthermore, patients were systematically queried about current medication use, and 8 pharmacotherapeutic groups were evaluated: lipid-lowering therapy, antihypertensives, oral hypoglycemic agents, insulin, thyroid hormone, uricosurics, psychoactive drugs, and nonsteroidal anti-inflammatory drugs. RESULTS: Sixty-six percent of the Spanish population was taking at least one medication. Therapeutic drug use was associated with age, independently of the higher prevalence of diabetes mellitus, hypertension, or hyperlipidemia in older patients. Sex disparities were found in the use of lipid-lowering agents, allopurinol, levothyroxine, nonsteroidal anti-inflammatory drugs, and psychoactive drugs. Use of psychoactive drugs was related to education level, work status, physical activity, smoking, and alcohol consumption. Almost 30% of patients with diabetes mellitus were taking 6 or more medications daily. Diabetes mellitus was associated with greater use of antihypertensives, lipid-lowering agents, and nonsteroidal anti-inflammatory drugs. CONCLUSIONS: Age and sex are the most important factors determining therapeutic drug use. Lifestyle patterns and sociocultural factors have an impact only on psychoactive drug use. Diabetes mellitus is associated with greater use of antihypertensives, lipid-lowering agents, and nonsteroidal anti-inflammatory drugs.

The UBA-UIM domains of the USP25 regulate the enzyme ubiquitination state and modulate substrate recognition.

USP25m is the muscle isoform of the deubiquitinating (DUB) enzyme USP25. Similarly to most DUBs, data on USP25 regulation and substrate recognition is scarce. In silico analysis predicted three ubiquitin binding domains (UBDs) at the N-terminus: one ubiquitin-associated domain (UBA) and two ubiquitin-interacting motifs (UIMs), whereas no clear structural homology at the extended C-terminal region outside the catalytic domains were detected. In order to asses the contribution of the UBDs and the C-terminus to the regulation of USP25m catalytic activity, ubiquitination state and substrate interaction, serial and combinatorial deletions were generated. Our results showed that USP25m catalytic activity did not strictly depend on the UBDs, but required a coiled-coil stretch between amino acids 679 to 769. USP25 oligomerized but this interaction did not require either the UBDs or the C-terminus. Besides, USP25 was monoubiquitinated and able to autodeubiquitinate in a possible loop of autoregulation. UBDs favored the monoubiquitination of USP25m at the preferential site lysine 99 (K99). This residue had been previously shown to be a target for SUMO and this modification inhibited USP25 activity. We showed that mutation of K99 clearly diminished USP25-dependent rescue of the specific substrate MyBPC1 from proteasome degradation, thereby supporting a new mechanistic model, in which USP25m is regulated through alternative conjugation of ubiquitin (activating) or SUMO (inhibiting) to the same lysine residue (K99), which may promote the interaction with distinct intramolecular regulatory domains.

Genetic analysis of p38 MAP kinases in myogenesis: fundamental role of p38alpha in abrogating myoblast proliferation.

The p38 mitogen-activated protein kinase (MAPK) pathway plays a critical role in skeletal muscle differentiation. However, the relative contribution of the four p38 MAPKs (p38alpha, p38beta, p38gamma and p38delta) to this process is unknown. Here we show that myoblasts lacking p38alpha, but not those lacking p38beta or p38delta, are unable to differentiate and form multinucleated myotubes, whereas p38gamma-deficient myoblasts exhibit an attenuated fusion capacity. The defective myogenesis in the absence of p38alpha is caused by delayed cell-cycle exit and continuous proliferation in differentiation-promoting conditions. Indeed, activation of JNK/cJun was enhanced in p38alpha-deficient myoblasts leading to increased cyclin D1 transcription, whereas inhibition of JNK activity rescued the proliferation phenotype. Thus, p38alpha controls myogenesis by antagonizing the activation of the JNK proliferation-promoting pathway, before its direct effect on muscle differentiation-specific gene transcription. More importantly, in agreement with the defective myogenesis of cultured p38alpha(Delta/Delta) myoblasts, neonatal muscle deficient in p38alpha shows cellular hyperproliferation and delayed maturation. This study provides novel evidence of a fundamental role of p38alpha in muscle formation in vitro and in vivo.