miR-23a, miR-24 and miR-27a protect differentiating ESCs from BMP4-induced apoptosis.

Numerous studies have indicated that BMP4 signaling is involved in the regulation of the early steps of development. In mouse embryonic stem cells (ESCs), BMP4 is crucial to sustain pluripotency and blocks differentiation towards neural fate. Here, through a systematic analysis of miRNAs in ESCs, we establish that BMP4 signaling regulates miR-23a, 27a and 24-2, through the recruitment of phospho-Smads at the promoter of the gene encoding this miRNA cluster. Suppression of miR-23a/b, 27a/b and 24 does not affect self-renewal or pluripotency, but induces an evident change of ESC differentiation, with a significant increase of the cells undergoing apoptosis after the transition from ESCs to epiblast stem cells (EpiSCs). BMP4 has been previously reported to cause apoptosis during ESC differentiation. By blocking BMP4 signaling, we completely prevent the apoptosis induced by suppression of the miRs. This suggests that the effects of miR suppression are the result of enhanced BMP4 signaling. This hypothesis is further supported by the observation that Smad5, the transcription factor downstream of the BMP4 receptor, is targeted by the miRNAs of the 23a and 23b clusters. Altogether, our results highlight the existence of a regulatory loop, involving Smad5 and the miR-23a clusters, that modulates the apoptotic response of ESCs to BMP4.

Antiretroviral treatment-based cost saving interventions may offset expenses for new patients and earlier treatment start.

OBJECTIVES: Combination antiretroviral therapy (cART) has become the main driver of total costs of caring for persons living with HIV (PLHIV). The present study estimated the short/medium-term cost trends in response to the recent evolution of national guidelines and regional therapeutic protocols for cART in Italy. METHODS: We developed a deterministic mathematical model that was calibrated using epidemic data for Lazio, a region located in central Italy with about six million inhabitants. RESULTS: In the Base Case Scenario, the estimated number of PLHIV in the Lazio region increased over the period 2012-2016 from 14 414 to 17 179. Over the same period, the average projected annual cost for treating the HIV-infected population was €147.0 million. An earlier cART initiation resulted in a rise of 2.3% in the average estimated annual cost, whereas an increase from 27% to 50% in the proportion of naïve subjects starting cART with a nonnucleoside reverse transcriptase inhibitor (NNRTI)-based regimen resulted in a reduction of 0.3%. Simplification strategies based on NNRTIs co-formulated in a single tablet regimen and protease inhibitor/ritonavir-boosted monotherapy produced an overall reduction in average annual costs of 1.5%. A further average saving of 3.3% resulted from the introduction of generic antiretroviral drugs. CONCLUSIONS: In the medium term, cost saving interventions could finance the increase in costs resulting from the inertial growth in the number of patients requiring treatment and from the earlier treatment initiation recommended in recent guidelines.

[Biography and bibliography of Francesco Fede, the founder of Italian pediatrics]. / Biografia e bibliografia di Francesco Fede, fondatore della pediatria italiana.

For the first time, an overall study of the life and works of Francesco Fede, the founder of Italian pediatrics, has been carried out. Unpublished biographical data was collected and the complete bibliographic works of Fede were presented. Fede is the hallmark of both scientific matter, which reached a climax in the definition of the Riga and Fede illness, and for his disinterested dedication as a Member of Parliament to foster development in didactics, research and assistance for pediatrics.

[Health-related quality of life in patients with chronic kidney disease]. / Qualita' della vita nei nefropatici.

Assessment of quality of life in patients with different degrees of chronic kidney disease is an important issue because of its impact on clinical decisions and financial resource management in the health-care system. The aim of this study was to assess whether a generic instrument like the SF-36 questionnaire is able to discriminate three different populations of patients with different degrees of renal disease (pre-ESRD, ESRD, TxR). Five hundred sixty-three patients from 12 Italian nephrology units completed the SF-36 scales by themselves. The results from these samples were compared with those from the general population. Univariate analysis and multivariate regression were used. The generic SF-36 questionnaire proved to be a powerful instrument to discriminate populations with different degrees of chronic renal failure. The quality of life of patients on dialysis is significantly worse than that of the normal population and other patients with less severe renal function impairment.

Comparison of two humidification systems for long-term noninvasive mechanical ventilation.

There is no consensus concerning the best system of humidification during long-term noninvasive mechanical ventilation (NIMV). In a technical pilot randomised crossover 12-month study, 16 patients with stable chronic hypercapnic respiratory failure received either heated humidification or heat and moisture exchanger. Compliance with long-term NIMV, airway symptoms, side-effects and number of severe acute pulmonary exacerbations requiring hospitalisation were recorded. Two patients died. Intention-to-treat statistical analysis was performed on 14 patients. No significant differences were observed in compliance with long-term NIMV, but 10 out of 14 patients decided to continue long-term NIMV with heated humidification at the end of the trial. The incidence of side-effects, except for dry throat (significantly more often present using heat and moisture exchanger), hospitalisations and pneumonia were not significantly different. In the present pilot study, the use heated humidification and heat and moisture exchanger showed similar tolerance and side-effects, but a higher number of patients decided to continue long-term noninvasive mechanical ventilation with heated humidification. Further larger studies are required in order to confirm these findings.

Ageing with HIV: newly diagnosed older adults in Italy.

The prevalence of HIV/AIDS among people in midlife and late adulthood has been increasing in Western countries over the last decade. We analyzed data from a prospective, observational multi-centre study on individuals newly diagnosed with HIV between January 2004 and March 2007 in 10 public counselling and testing sites in Latium, Italy. At diagnosis, routine demographic, epidemiological, clinical and laboratory data are recorded, and patients are asked to complete a questionnaire investigating socio-demographic and psycho-behavioural aspects. To analyze the association of individual characteristics with age, we compared older adults (> or = 50 years) with their younger counterpart (18-49 years). To adjust for potential confounding effect of the epidemiological, clinical and behavioural characteristics, to identify factors associated with older age at HIV diagnosis, multivariate logistic regression analysis was performed. Overall, 1073 individuals were identified, 125 of whom (11.6%) were aged 50 years or above. The questionnaire was completed by 41% (440/1073). Compared with their younger counterparts, a higher proportion of older patients were males, born in Italy, reported heterosexual or unknown HIV risk exposure, were never tested for HIV before and were in a more advanced stage of HIV infection at diagnosis. In addition, older adults had a lower educational level and were more frequently living with their partners or children. With respect to psycho-behavioural characteristics, older patients were more likely to have paid money for sex and have never used recreational drugs. Interestingly, no differences were found regarding condom use, which was poor in both age groups. These findings may have important implications for the management of older adults with HIV, who should be targeted by appropriate public health actions, such as opportunistic screening and easier access to healthcare. Moreover, strategies including information on HIV and prevention of risk behaviours are needed.

Current evidence on diagnostic accuracy of commercially based nucleic acid amplification tests for the diagnosis of pulmonary tuberculosis.

BACKGROUND: Even though commercial nucleic acid amplification tests (NAATs) have become the most frequently used molecular tests for laboratory diagnosis of pulmonary tuberculosis (TB), published studies report variable estimates of their diagnostic accuracy. We analysed the accuracy of commercial NAATs for the diagnosis of pulmonary TB in smear positive and smear negative respiratory samples using culture as a reference standard. METHODS: English language studies reporting data sufficient for calculating sensitivity and specificity of commercial NAATs on smear positive and/or smear negative respiratory samples were included. Meta-regression was used to analyse associations with reference test quality, the prevalence of TB, sample and test type. Predictive values for different levels of pre-test probability were quantified using Bayes' approach. RESULTS: Sixty three journal articles published between 1995 and 2004 met the inclusion criteria. Pooled sensitivity and specificity were 0.96 and 0.85 among smear positive samples and 0.66 and 0.98 among smear negative samples. The number of culture media used as reference test, the inclusion of bronchial samples, and the TB prevalence were found to influence the reported accuracy. The test type had no effect on the diagnostic odds ratio but seemed to be correlated with sensitivity or specificity, probably via a threshold effect. CONCLUSIONS: Commercial NAATs can be confidently used to exclude TB in patients with smear positive samples in which environmental mycobacteria infection is suspected and to confirm TB in a proportion of smear negative cases. The methodological characteristics of primary studies have a considerable effect on the reported diagnostic accuracy.

[Knowledge and use of Pap-smear among HIV-positive women]. / Importanza della conoscenza del Pap-test e suo utilizzo tra le donne HIV-positive.

AIM: HIV-positive women are at increased risk for preneoplastic lesions and invasive cervical cancer (ICC). The occurrence of these lesions can be substantially reduced by appropriate cervico-vaginal screening protocols (i.e., Pap-test). The aim of study was to assess: 1) awareness of Pap-smear and 2) the association between awareness of Pap-smear and screening attitudes of HIV-positive women. METHODS: Three-hundred and ninety HIV-positive women who attended the HIV outpatient gynecological unit of the National Institute for Infectious Diseases, Rome, from January 2003 to April 2005 were included in this investigation. These 390 women were interviewed to assess whether they were aware that Pap-test was a preventive tool against cervical cancer. In addition, past history of Pap-test, socioeconomic condition, history of HIV infection, and sexual habits were investigated. Odds ratios (OR) and 95% confidence intervals (CI) were used to assess the association between knowledge of Pap-test and covariates. RESULTS: Of these 390 HIV-positive women, 54.6% were not aware that Pap-test could prevent ICC. Women with a low educational level (OR = 6.6) or women who originated from Africa (OR = 6.5) were more likely to be unaware of Pap-test. Lack of Pap-test awareness was strongly associated with negative history for lifetime Pap-test (OR = 4.7). CONCLUSIONS: We showed that a large proportion of HIV-infected women are not aware that ICC could be prevented through Pap-test screening, and that lack of Pap-test screening is strongly associated with lack of awareness. The need for Pap-test counseling targeted to HIV-infected women clearly emerges from our findings.

Infection with human herpesvirus type 8 and Kaposi's sarcoma in Sardinia.

BACKGROUND: A cross-sectional study was conducted in the provinces of Sassari (northern Sardinia, covered by a population-based cancer registry), and of Cagliari (southern Sardinia) to estimate the prevalence of infection with human herpesvirus type 8 (HHV8) and the incidence of classic Kaposi's sarcoma (KS) among HHV8-infected individuals. PATIENTS AND METHODS: Sera from 297 hospitalized persons potentially at risk of developing classic KS (i. e., those aged 50 years or older) were tested for antibodies against HHV8. HHV8 seroprevalence rates (with 95% confidence intervals-CI) and yearly incidence rates (IR/100,000) of KS were calculated according to age and sex. RESULTS: Of tested individuals, 32.0% had antibodies against HHV8 in Sassari and 30.0% in Cagliari. Estimated IR of KS among HHV8-positive persons and KS:HHV8 ratio were two times higher in Sassari (1:3,891) than in Cagliari (1:8,114), and higher in men (1:2,846 in Sassari; 1:5,483 in Cagliari) as compared to women (1:6,827 in Sassari; 1:12,489 in Cagliari). CONCLUSIONS: Although the overall prevalence of HHV8 seemed similar in Sassari and in Cagliari, the risk of KS was higher in Sassari, suggesting that different cofactor(s), or different distribution of the same cofactor(s) between the two provinces of Sardinia, might have played a role in KS development.

MRSA carriage: the relationship between community and healthcare setting. A study in an Italian hospital.

From May 1997 to June 1998, all patients admitted to the study institution were screened at entry for MRSA carriage (both colonization and infection). Eighty-six MRSA carriers were identified; of these, 85 were nasal carriers. Risk factors were compared to those of 86 controls. Although the vast majority of both carriers and controls had at least one previous hospital stay, carriers were less likely than controls to be referred from a community setting, and had resided within the community for a shorter time before the current admission. The number of underlying conditions was comparable in the two groups, but those infected were more likely to have cancer than the controls. While community-acquired MRSA carriage is rare, exposure to a health care setting (particularly if repeated) within six months from the current admission, is a risk factor for MRSA carriage and introduction of the organism into an institution.

Activity of daptomycin against enterococci and coagulase-negative staphylococci (CNS): relationship between CNS susceptibility and slime production.

We compared the in vitro activity of daptomycin, a new lipopeptide antibiotic, with that of vancomycin and other selected agents against 95 coagulase negative staphylococci (CNS) isolates causing septicemia or foreign-body infections in immunocompromised patients. These strains were classified as follows: 51 methicillin-susceptible CNS (23 slime producers); 44 methicillin-resistant CNS (23 slime producers). We also investigated the activity of daptomycin against 50 Enterococcus faecalis isolates. Minimal inhibitory concentrations (MICs) were determined by the broth microdilution method. Daptomycin at a concentration of 2 mg/L was inhibitory for all the evaluated strains. Vancomycin and ciprofloxacin showed good activity: 90% of the strains were inhibited by these agents at 8 mg/L. The activity of netilmicin, rifampin and trimethoprim-sulfamethoxazole was instead limited. Resistance to the antimicrobial agents tested was seen with increased frequency among slime producing strains. Daptomycin and teicoplanin were the most active agents tested against E. faecalis (MIC90 0.25 mg/L and 0.12 mg/L).

Evaluation of intraventricular teicoplanin for the treatment of neurosurgical shunt infections.

Seven patients with staphylococcal neurosurgical shunt infections were treated with intraventricular teicoplanin. Two infants received 5 mg/d, three patients received 20 mg/d, and two patients received 20 mg every other day. Six of these patients also received intravenous antibiotics. Three patients had infections caused by methicillin-susceptible Staphylococcus epidermidis, and one patient had an infection caused by methicillin-resistant S. epidermidis. Three patients were infected with Staphylococcus aureus (one with a methicillin-resistant strain and two with methicillin-susceptible strains). The mean duration of intraventricular therapy was 16 days. Sterilization of cerebrospinal fluid (CSF) was obtained after an average of 4.4 days. All patients were cured both clinically and microbiologically. No significant adverse effects were observed in any patients. Penetration of teicoplanin into the CSF after intravenous administration was poor. However, after intraventricular administration, high and prolonged peak and trough levels of teicoplanin were detected in the CSF. Bactericidal activity of the CSF was remarkable, exceeding the 1:8 dilution in the majority of the cases. The alternate-day schedule of intraventricular administration of teicoplanin was as effective as the once-daily regimen.

Norfloxacin versus cotrimoxazole for infection prophylaxis in granulocytopenic patients with acute leukemia. A prospective randomized study.

Norfloxacin (NOR) or cotrimoxazole (TMP/SMX) were randomly administered to 59 granulocytopenic patients with acute leukemia for prevention of bacterial infections. Nineteen NOR patients (65%) and 22 TMP/SMX patients (73%) complained of febrile or infectious episodes during the study. The mean incidence of febrile complications per patient was higher in the TMP/SMX group: 1.05 vs 0.68 (p less than 0.05). Eleven of 16 microbiologically documented infections in the TMP/SMX group and 7 of 11 in the NOR group were caused by gram negative bacilli (GNB). NOR recipients had fewer days of fever, fewer days on parenteral antibiotics and a lower proportion of time spent febrile. Fecal surveillance cultures showed intestinal GNB colonization in 42/80 specimens in the TMP/SMX group (resistant strains: 93%) and in 8/75 specimens in the NOR group (1 resistant strain). Overall, NOR seems to be effective in eradicating GNB from the digestive tract without selection of resistant strains and in preventing febrile episodes in neutropenic patients.

Antibacterial activity of combined cefotaxime and desacetyl-cefotaxime against aerobic and anaerobic gram-negative bacilli.

Synergy between cefotaxime and desacetyl-cefotaxime was evaluated against 12 strains (9 Enterobacteriaceae and 3 Bacteroides fragilis) by the chequerboard technique. A 1:1 combination of cefotaxime and desacetyl-cefotaxime was synergistic against two-thirds of the 12 strains tested. The in vitro activity of the combination was compared with that of four other beta-lactam antibiotics against 96 recent clinical isolates: 78 Enterobacteriaceae, 8 Haemophilus influenzae, 10 B. fragilis. The MICs of the combination for Gram-negative bacilli were similar to those of ceftazidime. Cefotaxime/desacetyl-cefotaxime was more active than cefotetan, cefonicid and piperacillin against Enterobacteriaceae and H. influenzae.

In vitro antibacterial activity of cefotaxime and desacetylcefotaxime, alone or in combination, against gram-positive cocci.

The in vitro activity of cefotaxime and desacetylcefotaxime against Staphylococcus aureus, Staph. epidermidis and Streptococcus pyogenes was investigated. Synergy studies were performed using time-kill curves and the chequerboard test. The time-kill curves were performed on five strains each of Staph. aureus, Staph. epidermidis and Strep. pyogenes; cefotaxime and desacetylcefotaxime were tested alone or in combination at MIC and sub-MIC values. The chequerboard test was performed in microtitre plates on ten strains each of Staph. aureus, Staph. epidermidis and Strep. pyogenes: the results were interpreted by the fractional inhibitory concentration index. In some cases both methods showed synergistic interaction against the staphylococci tested. Indifference was observed against Strep. pyogenes.

Prophylactic co-trimoxazole versus norfloxacin in neutropenic children--perspective randomized study.

Co-trimoxazole or norfloxacin were randomly administered to 44 granulocytopenic children with malignancies in order to prevent bacterial infections. Although more patients in the co-trimoxazole group had febrile episodes (p less than 0.01), the mean of febrile days and the mean of days with systemic antibiotics did not differ significantly in the two groups. Five patients in the co-trimoxazole group had a microbiologically documented infection (four with septicemia) due to Escherichia coli (n = 2), Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus sp. There were four septicemic episodes in the norfloxacin group due to P. aeruginosa, Streptococcus pneumoniae, Streptococcus mitis and Streptococcus faecalis. Compliance was good during administration of both drugs. No signs or symptoms of arthropathy were seen in the norfloxacin group. The number of gram-negative bacilli resistant to co-trimoxazole isolated from stools significantly increased during prophylaxis with co-trimoxazole (p less than 0.001). Norfloxacin did not select resistant strains and was very active in eradicating gram-negative bacilli from stools (27.5% of positive cultures).

Serum bactericidal activity against aerobes and anaerobes of volunteers receiving cefoxitin or cefotetan.

Six volunteers received intravenously a single 1 g dose of cefoxitin or cefotetan. The 2 groups were crossed after a week of washout. Five strains each of Escherichia coli, Klebsiella pneumoniae, Bacteroides fragilis and Bacteroides thetaiotaomicron susceptible to the administered drugs were tested for serum bactericidal activity (SBA). Blood samples were obtained before and 0.5, 3.0 and 12.0 hours after antibiotic injection. SBA was determined using microtitre procedures. Anaerobic bacteria were incubated in an anaerobic chamber. Cefotetan showed a very high SBA both against aerobes and anaerobes over the 12 hour sampling time. Cefoxitin reached satisfactory SBA values only 0.5 hours after administration.

Antibacterial activity after a single-dose of norfloxacin, ofloxacin and pipemidic acid detected in urine of volunteers.

Six healthy volunteers received in a triple-crossover design a single oral dose of norfloxacin, ofloxacin or pipemidic acid. Urine samples were collected during the 24 h following the administration and were tested for inhibitory and bactericidal activity against selected strains of Enterobacteriaceae and Pseudomonas aeruginosa. Norfloxacin and ofloxacin inhibited the urinary growth of sensitive strains during the 24 h of sampling time at dilutions much higher than those generally considered satisfactory. Nalidixic acid was less effective and did not achieve bactericidal activity against Ps. aeruginosa over the interval of 12 to 24 h.