Active screening of COVID-19-associated pulmonary aspergillosis with serum beta-glucan and endotracheal aspirates galactomannan and fungal culture.

BACKGROUND: Since February 2021 active screening of COVID-19-associated pulmonary aspergillosis (CAPA) has been implemented in our institution. OBJECTIVES: To evaluate CAPA incidence in our centre and evaluate performance of our screening protocol. METHODS: We screened once per week, collecting endotracheal aspirates for fungal culture and galactomannan (GM) and serum for 1,3-ß-D-glucan (BG). In case of positivity (GM more than 4.5, platelia assay, and/or BG >7 pg/ml, wako and/or positive fungal culture), second-level investigations were performed to pursue CAPA diagnosis according to ECMM/ISHAM criteria: bronchoalveolar lavage (BAL) fungal culture and GM, chest computed tomography (CT), serum GM. RESULTS: A total of 102 patients were screened (median age 64 years, range 39-79; 28 (27.4%) females). Twenty-two patients were diagnosed with CAPA (21%). 12 patients were positive for serum BG, 17 patients were positive for endotracheal aspirates GM and 27 patients were positive for endotracheal aspirates fungal culture. Thirty-two BALs were performed, and 26 patients underwent CT chest. Following the second level investigations 61% of the patients with positive screening tests were diagnosed with CAPA. Serum BG above 20 pg/ml or positive serum GM were always associated with typical CT chest signs of aspergillosis. Compared with 1 single positive test, having 2 positive screening test was significantly more associated with CAPA diagnosis (p = .0004). CONCLUSIONS: Active CAPA screening with serum 1,3-ß-D-glucan and endotracheal aspirates galactomannan and fungal cultures and consequent second level investigations led to high number of CAPA diagnosis. Combining more positive fungal biomarkers was more predictive of CAPA diagnosis.

Second Trimester Fetal Loss Due to Citrobacter koseri Infection: A Rare Cause of Preterm Premature Rupture of Membranes (PPROM).

Citrobacter koseri is a facultative anaerobic, motile, non-spore-forming Gram-negative bacillus, which belongs to the family of Enterobacteriaceae. Severe infections due to Citrobacter spp. have been reported in the urinary tract, respiratory airways, intra-abdominal organs, skin and soft tissue, eye, bone, bloodstream, and central nervous system. In newborns, C. koseri is a well-known cause of meningitis, cerebral abscesses, brain adhesions, encephalitis, and pneumocephalus. Infection can be acquired through vertical maternal transmission or horizontal hospital settings; however, in many cases, the source is unknown. Preterm premature rupture of membranes (PPROM), caused by C. koseri, has rarely been described. Herein, we describe a case of PPROM at 16 weeks and 3 days of gestation, leading to anhydramnios. The parents opted for legal termination of the pregnancy, as the prognosis was very poor. C. koseri was isolated postmortem from a placental subamniotic swab and parenchymal sample, as well as fetal blood and lung. To the best of our knowledge, this is the first case of early second-trimester PPROM in which C. koseri infection was demonstrated.

Diagnostic approach and epidemiology of Microbial Keratitis: findings from an Italian Tertiary Care center.

Rapid identification of causative microorganisms of microbial keratitis (MK) and knowledge of the most common local pathogens are prerequisites for rational antimicrobial therapy. We retrospectively reviewed the characteristics of MK diagnosed at the IRCCS Arcispedale Santa Maria Nuova of Reggio Emilia (Italy) in a 5-years period, where the Ophthalmologist Unit is a reference center for corneal infections. During the study period, 183 MK were evaluated through corneal scrapings cultures. The positivity rate was 54,1%. A total of 107 microorganisms have been isolated: Acanthamoeba species was the etiologic agent in 19 cases. Pseudomonas aeruginosa and Staphylococcus aureus were more frequently isolated in bacterial keratitis, while Fusarium spp., Candida albicans, and Alternaria alternata were predominant among the fungal isolates. Strict cooperation between ophthalmologists and clinical microbiologists is advisable to allow the best diagnostic approach for MK.

Klebsiella pneumoniae Chorioamnionitis: An Underrecognized Cause of Preterm Premature Rupture of Membranes in the Second Trimester.

Klebsiella pneumoniae is a Gram-negative, rod-shaped bacterium, responsible for hospital and community acquired pneumonia, urinary tract and wound infections, and bloodstream dissemination. K. pneumoniae infection in pregnancy, leading to acute chorioamnionitis (AC), preterm premature rupture of membranes (PPROM) and early pregnancy loss in the second trimester, has been rarely reported. Herein, we present a case of K. pneumoniae AC that caused intrauterine fetal demise (IUFD) at 19 weeks + 5 days. The 36-year-old mother was admitted at 18 weeks + 1 day of gestation for threatened abortion. IUFD occurred 11 days after. Fetal postmortem showed severe AC and funisitis, neutrophils within alveoli and intestinal lumen, associated with rod-like bacteria. Fetal blood and lung cultures grew K. pneumoniae, ß-lactamase-non-producing strain. Antibiogram revealed sensitivity for piperacillin/tazobactam. Three days after IUFD, the mother presented with fever (37.8 °C) which persisted for one week. Maternal blood and urine cultures were negative. According to fetal microbiological results, available 6 days after IUFD, initial treatment with amoxicillin/clavulanic acid was replaced with piperacillin/tazobactam with full patient recovery. Therefore, in the event of PPROM and IUFD, fetal microbiological investigations should always be performed to isolate the proper etiologic agent and start the correct medical treatment.

Lifetime growth and risk of testicular cancer.

Adult height is associated with testicular cancer risk. We studied to what extent this association is explained by parental height, childhood height and age at puberty. We conducted a case-control study on germ-cell testicular cancer patients diagnosed in 1997-2008 and resident in the Province of Turin. Information was collected using mailed questionnaires in 2008-2011. Specifically, we asked for adult height (in cm), height at age 9 and 13 (compared to peers) and age at puberty (compared to peers). We also asked for paternal and maternal height (in cm) as indicators of genetic components of adult height. The analysis included 255 cases and 459 controls. Odds ratios (ORs) of testicular cancer were estimated for the different anthropometric variables. Adult height was associated with testicular cancer risk [OR: 1.16, 95% confidence interval (CI): 1.03-1.31 per 5-cm increase]. The risk of testicular cancer was only slightly increased for being taller vs. shorter than peers at age 9 (OR: 1.55, 95% CI: 0.91-2.64) or age 13 (OR: 1.26, 95% CI: 0.78-2.01), and parental height was not associated with testicular cancer risk. The OR for adult height was 1.32 (95% CI: 1.12-1.56) after adjustment for parental height. Among participants with small average parental height (<167 cm or less), the OR of testicular cancer for tall (>180 cm) vs. short (<174 cm) subjects was 3.47 (95% CI: 1.60-7.51). These results suggest that the association between height and testicular cancer is likely to be explained by environmental factors affecting growth in early life, childhood and adolescence.

Experimental biofilm-related Candida infections.

AIM: We investigated the pathogenic role of biofilm and the therapeutic efficacy of anidulafungin in experimental infections of the wax moth Galleria mellonella by Candida albicans clinical strains. MATERIALS & METHODS: On the basis of the in vitro propensity to form biofilm, five biofilm-producer (BP) and four nonproducer (NP) C. albicans clinical strains were used in this study. For each strain, we assessed the virulence by infecting G. mellonella larvae and observing survival. Anidulafungin was administered 2 h after yeast inoculum at 0.6 µg, according to the therapeutic dose recommended for humans. RESULTS: Biofilm-forming ability highly influenced the larva-killing rate. A significant (p < 0.0001) survival decrease was observed in the BP group, with 80% of the infected larvae dying within 72 h. NP isolates did not reach the same killing rate, even at the end of experiments (216 h). Larval survival was enhanced (p < 0.0001) by anidulafungin administration in both groups. Survival rate at 72 h was similar in both groups (BP 78.5% and NP 87.5%); whereas there were still differences at the end of the experiments, with a higher survival in the NP group (75 vs 48%). CONCLUSION: Our data confirm the pathogenic role of biofilm in C. albicans infections. Its importance was further enhanced by a lack of contribution from extracellular enzymes, detected in both NP and BP strains. In addition, we demonstrated anidulafungin efficacy in treating biofilm-related invasive candidiasis.

DNA methyltransferase 3b (DNMT3b), tumor tissue DNA methylation, Gleason score, and prostate cancer mortality: investigating causal relationships.

PURPOSE: Aberrant DNA methylation plays a role in prostate cancer progression. We studied the relationships among DNA methyltransferase (DNMT) genotype, DNA methylation, Gleason score, and mortality in two cohorts of prostate cancer patients, previously reported with associations between DNA methylation in GSTP1, APC, and RUNX3 and prostate cancer mortality. Herein, we considered possible causal relationships between the studied variables, assuming that (1) DNMT activity affects tumor tissue methylation, (2) methylation status affects tumor morphology, and thus the Gleason score, and (3) DNA methylation affects mortality via Gleason score. METHODS: The cohorts comprised 438 patients diagnosed at one Italian pathology ward before 1997, with DNA obtained from paraffin-embedded tumor tissues. The polymorphism rs406193 in the DNMT3b gene was assessed by allele discrimination in real-time PCR. According to the assumed causal model, we analyzed the effects of rs406193 (T carriers vs others) on the Gleason score without adjusting for gene methylation, and the effects of rs406193 on gene methylation and prostate cancer mortality without adjusting for Gleason score. RESULTS: We found no evidence of association between T carriers and the number of methylated genes. However, T carriers had reduced risk of a Gleason score 8+ (odds ratio = 0.57, 95 % CI 0.39-0.85), and a hazard ratio of 0.81 (0.61-1.09) of dying from prostate cancer, which would have been erroneously estimated of 0.93 if adjusted for Gleason score. CONCLUSIONS: These findings provide clues on the role of a DNMT3b SNP in prostate cancer progression and illustrate the importance of considering possible causal relationships in the analyses.

Effects of Italian smoking regulation on rates of hospital admission for acute coronary events: a country-wide study.

BACKGROUND: Several studies have reported a reduction in acute coronary events (ACEs) in the general population after the enforcement of smoking regulations, although there is uncertainty concerning the magnitude of the effect of such interventions. We conducted a country-wide evaluation of the health effects of the introduction of a smoking ban in public places, using data on hospital admissions for ACEs from the Italian population after the implementation of a national smoking regulation in January 2005. METHODS AND FINDINGS: Rates of admission for ACEs in the 20 Italian regions from January 2002 to November 2006 were analysed using mixed-effect regression models that allowed for long-term trends and seasonality. Standard methods for interrupted time-series were adopted to assess the immediate and gradual effects of the smoking ban. Effect modification by age was investigated, with the assumption that exposure to passive smoking in public places would be greater among young people. In total, 936,519 hospital admissions for ACEs occurred in the Italian population during the study period. A 4% reduction in hospital admissions for ACEs among persons aged less than 70 years was evident after the introduction of the ban (Rate Ratio [RR], 0.96; 95% Confidence Interval [CI], 0.95-0.98). No effect was found among persons aged at least 70 years (RR 1.00; 95% CI 0.99-1.02). Effect modification by age was further suggested by analyses using narrower age categories. CONCLUSIONS: Smoke-free policies can constitute a simple and inexpensive intervention for the prevention of cardiovascular diseases and thus should be included in prevention programmes.

It is just a game: lack of association between watching football matches and the risk of acute cardiovascular events.

BACKGROUND: The role of trigger factors in acute cardiovascular events has been much studied in the past few years. A recent study analysed changes in the rates of cardiac emergencies in Bavaria (Germany) during the last Football World Cup. The authors reported a 2.7-fold increase in the incidence of cardiac emergencies in the 12 h before and after football matches involving the German team, which sparked the debate on the necessity of the introduction of ad hoc cardiovascular preventive measures. METHODS: We studied 25,159 hospital admissions for acute myocardial infarction (AMI) among the Italian population during three international football competitions: the World Cup 2002, the European Championship 2004 and the World Cup 2006. Poisson regression was used to estimate the relative risk of hospital admission for AMI on the days when football matches involving the Italian team were disputed, compared with the other days of the three competitions. Furthermore, we reviewed the available published studies regarding the association between football matches and the risk of cardiovascular events. RESULTS: We did not find an increase in the rates of admission for AMI on the days of football matches involving Italy in either the single competitions or the three competitions combined (relative risk 1.01; 95% confidence interval 0.98-1.05). We identified 10 studies published on this topic. With the exception of the recently published German study and two small Swiss studies, all relative risk estimates were between 0.7 and 1.3. CONCLUSIONS: The cardiovascular effects of watching football matches are likely to be, if anything, very small.

Promoter methylation in APC, RUNX3, and GSTP1 and mortality in prostate cancer patients.

PURPOSE There is a need to better understand prostate cancer progression and identify new prognostic markers for this tumor. We investigated the association between promoter methylation in a priori selected genes and survival in two independent large series of prostate cancer patients. METHODS We followed up with two cohorts of patients (216 patients diagnosed in 1982 to 1988 and 243 patients diagnosed in 1993 to 1996) diagnosed at one hospital pathology ward in Turin, Italy. DNA was obtained from paraffin-embedded tumor tissues and evaluated for promoter methylation status in glutathione S-transferase (GSTP1), adenomatous polyposis coli (APC), and runt-related transcription factor 3 (RUNX3). Results The two cohorts had different prevalences of methylation in APC (P = .047), GSTP1 (P = .002), and RUNX3 (P < .001). Methylation in APC was associated with an increased risk of prostate cancer-specific mortality (hazard ratio [HR] = 1.42; 95% CI, 0.98 to 2.07 in the 1980s cohort; HR = 1.57; 95% CI, 0.95 to 2.62 in the 1990s cohort; HR = 1.49; 95% CI, 1.11 to 2.00 in the two cohorts combined). In subgroup analyses, the HRs were higher among patients with a Gleason score less than 8 (HR = 1.52; 95% CI, 0.85 to 2.73 in the 1980s cohort; HR = 2.09; 95% CI, 1.02 to 4.28 in the 1990s cohort). Methylation in RUNX3 was associated with prostate cancer mortality only in the 1990s cohort, and methylation in GSTP1 did not predict mortality in either cohort. CONCLUSION The pattern of hypermethylation may have changed after the introduction of prostate-specific antigen testing in the beginning of the 1990s. Promoter methylation in APC was identified as a marker for prostate cancer progression.

GP5+/6+ SYBR Green methodology for simultaneous screening and quantification of human papillomavirus.

BACKGROUND: Detection and quantification of human papillomavirus (HPV) may help in predicting the evolution of HPV infection and progression of associated lesions. OBJECTIVES: We propose a novel protocol using consensus primers GP5+/6+ in a SYBR Green quantitative real-time (Q-RT) polymerase chain reaction (PCR). The strategy permits screening for HPV infection and viral load quantification simultaneously. STUDY DESIGN: DNA from 153 archived cervical samples, previously tested for HPV detection by GP5+/6+ PCR and typed by EIA-RLB (enzyme immunoassay-reverse line blot) or sequence analysis, was analysed using SYBR Green Q-RT PCR. Melting temperature assay (T(m)) and cycle threshold (C(t)) were used to evaluate HPV positivity and viral load. The T(m) in the range of 77-82 degrees C was considered to be positive for HPV-DNA. HPV results generated through GP5+/6+ conventional PCR were considered the gold standard against which sensitivity and specificity of our assay were measured. RESULTS: Out of 104 HPV positive samples, 100 (96.2%) were also determined as positive by SYBR Green Q-RT PCR; of the 49 HPV-negative samples, all were determined as negative. There was an excellent positivity agreement (kappa=0.94) between the SYBR Green Q-RT and the previous methods employed. The specificity and sensitivity were 100% and 96.2%, respectively. Comparison of SYBR Green Q-RT and TaqMan oligo-probe technologies gave an excellent concordance (rho(c)=0.95) which validated the proposed strategy. CONCLUSIONS: We propose a sensitive and easy-to-perform technique for HPV screening and viral load quantification simultaneously.

How the effect of maternal age on the risk of childhood leukemia changed over time in Sweden, 1960-2004.

BACKGROUND: Previous studies on the association between maternal age and risk of childhood leukemia found inconsistent results. OBJECTIVES: We aimed to assess whether there is an association between maternal age and risk of childhood leukemia and whether such an association is modified by maternal year of birth. METHODS: By linking nationwide Swedish registers, we analyzed leukemia incidence among all children between 1 and 5 years of age born between 1960 and 1999. We estimated incidence time trends by child year of birth (overall and stratified by maternal age) and incidence rate ratios (RRs) for maternal age groups stratified by maternal birth cohort. We tested the interaction between maternal age and child year of birth through the likelihood ratio test between nested Poisson regression models. RESULTS: We observed 1,562 leukemia cases. The overall annual percent change (APC) was 1.00 [95% confidence interval (CI), 0.51 to 1.49]. Stratifying by maternal age classes, APCs decreased from 1.66 (0.68 to 2.65) for mothers or= 35 years at delivery. RRs for children born to the oldest with respect to the youngest mothers were 2.42 (1.31 to 4.67), 1.68 (1.00 to 2.72), 1.34 (0.87 to 2.01), and 0.87 (0.46-1.54) for mothers born in 1930-1934, 1940-1944, 1950-1954, and 1960-1964, respectively. CONCLUSIONS: Childhood leukemia risk increased with maternal age for mothers born in the past, whereas maternal age had no effect on this risk for mothers born more recently. This finding may explain the inconsistency of previous studies and suggests that leukemia risk may be related to an environmental factor to which women's exposure has changed over time.

Cardiovascular benefits of smoking regulations: The effect of decreased exposure to passive smoking.

OBJECTIVES: Decreases in population rates of acute myocardial infarction (AMI) have been repeatedly seen in the first months after the introduction of regulations banning smoking in public places. By decreasing the exposure to passive smoking and its acute cardiovascular effects, smoking regulations may cause an immediate drop in AMI incidence, beginning from the initial days after their introduction. Using simulations, we quantified this expected decrease. METHODS: Expected AMI decreases were estimated using several parameters, including prevalence of exposure to passive and active smoking, relative risks of AMI associated with active and passive smoking, decrease in exposure to passive smoking after the introduction of the regulation, and level of susceptibility of active smokers to exposure to passive smoking. RESULTS: After evaluating several possible combinations of these parameters, we found that AMI reductions of 5-15% seem likely. For example, a scenario with a population average of 5 hours per week of overall exposure to passive smoking, a 50% reduction in this exposure after the introduction of the regulation, an acute relative risk (RR) of AMI of 4.5 in the first hour after passive smoking exposure, and similar susceptibility to passive smoking among both active and passive smokers translates into a 8.6% decrease in AMI. CONCLUSIONS: Smoking regulations can cause immediate and relevant decreases in AMI through the prevention of exposure to passive smoking.

Feasibility of recruiting a birth cohort through the Internet: the experience of the NINFEA cohort.

The NINFEA cohort is an Italian birth cohort aiming at recruiting pregnant women through the Internet and following-up their children. To understand whether Internet-based recruitment was feasible we started a pilot in July 2005 targeted to pregnant women visiting the hospitals of the city of Turin (900,000 inhabitants), where we advertised the study. For this purpose we constructed a website (www.progettoninfea.it), with on-line questionnaires to be completed during pregnancy and at 6 and 18 months after delivery. Participants' characteristics were compared with those of women giving birth in Turin, which are routinely released as individual anonymous records. We also compared complete with partial respondents. We also carried out a survey of 122 women giving birth in the main Turin obstetric hospital to estimate the proportion of pregnant women with access to the Internet and awareness of the NINFEA cohort. By December 2006, we had recruited 670 women. Participation was associated with being older, a university graduate, primiparous and born in Italy. Complete response (n = 633) was associated with being primiparous and participation after the first trimester. In the survey, 66% (95% confidence interval: 56-74%; n = 80) of the women had access to the Internet and 42% (33-51%; n = 51) were aware of the study; 6.5% (2.9-12.5%; n = 8) had participated in the NINFEA cohort. Our study indicates that recruitment of an Internet-based birth cohort is feasible. As with many other types of birth cohort study, the participants are a self-selected sample. To minimise selection bias analyses should therefore be limited to internal comparisons.

Short-term effects of Italian smoking regulation on rates of hospital admission for acute myocardial infarction.

AIMS: We used the hospital discharge records of Piedmont region (northern Italy) to evaluate whether a national law banning smoking in public resulted in a short-term reduction in hospital admissions for acute myocardial infarction (AMI). METHODS AND RESULTS: Rates of admission for AMI before the ban (October-December 2004) and during the ban (February-June 2005) were analysed. Each period was compared with the corresponding period 12 months before. Among persons aged under 60, the number of admissions for AMI decreased significantly after the introduction of the ban: from 922 cases in February-June 2004 to 832 cases in February-June 2005 (sex- and age-adjusted rate ratio, 0.89; 95% confidence interval, 0.81-0.98). No decrease was seen before the ban. No effect was found among persons aged at least 60. We estimated that the observed reduction in active smoking after the introduction of the ban could account for a 0.7% decrease in admissions for AMI during the study period, suggesting that most of the observed effect (11%) might be due to the reduction of passive smoking. CONCLUSION: Our study, based on a population of about 4 million inhabitants, suggests that smoke-free policies may result in a short-term reduction in admissions for AMI.