Variable performance of different commercial systems for testing carbapenem susceptibility of KPC carbapenemase-producing Escherichia coli.

OBJECTIVES: The aim was to evaluate different methods for testing carbapenem susceptibility of Escherichia coli producing KPC-type carbapenemase. METHODS: Susceptibility to imipenem, meropenem and ertapenem was assayed using the reference broth microdilution method and several commercial methods (Vitek2, MicroScan, Etest, MIC Test Strip) starting from the same bacterial suspension. Susceptibility to imipenem and meropenem was also tested by Sensititre and disc diffusion (Bio-Rad). Results were interpreted according to EUCAST clinical breakpoints. Essential agreement (EA), category agreement (CA) and error rates were calculated as described by the International Organization for Standardization (ISO) guidelines and also considering the new EUCAST definitions. Genotypic diversity of isolates was evaluated with a RAPD profiling protocol. RESULTS: Of 54 KPC-positive E. coli isolates, 5.6%, 7.4% and 0% were susceptible standard dosing regimen (S), 55.6%, 72.2% and 0% susceptible increased exposure (I), and 38.9%, 20.4% and 100.0% resistant (R) to imipenem, meropenem and ertapenem, respectively, using the reference broth microdilution method. CA lower than 90% were observed with all systems for imipenem and meropenem using both the ISO and the modified EUCAST criteria. With ertapenem, CA >90% was observed with all methods except Vitek2. RAPD profiling revealed a remarkable genotypic diversity of the isolates, supporting that results were not biased by an oligoclonal nature of the collection. CONCLUSIONS: Commercial methods can be unreliable for testing susceptibility to carbapenems of KPC-producing E. coli. Susceptibility should be confirmed by reference broth microdilution.

Colonization by multidrug-resistant organisms in long-term care facilities in Italy: a point-prevalence study.

OBJECTIVES: To determine prevalence and risk factors for colonization by multidrug-resistant organisms (MDROs) in long-term care facility (LTCF) residents in Italy. Genotypes of MDRO isolates were investigated. METHODS: A point-prevalence study was conducted at 12 LTCFs located in four Italian cities (2 February to 14 March 2015). Rectal swabs, faeces and nasal/auxiliary swabs were cultured for extended-spectrum ß-lactamase (ESBL)- and/or carbapenemase-producing Enterobacteriaceae, Clostridium difficile and methicillin-resistant Staphylococcus aureus (MRSA) respectively. Antimicrobial susceptibility testing, detection of ESBL and/or carbapenemase genes and molecular typing of MDROs were performed. Risk factors for colonization were determined by univariate and multivariate analysis. RESULTS: A total of 489 LTCF residents aged ≥65 years were enrolled. The prevalence of colonization by ESBL-producing Enterobacteriaceae, MRSA and C. difficile was 57.3% (279/487), 17.2% (84/487) and 5.1% (21/409) respectively. Carriage rate of carbapenemase-producing Enterobacteriaceae was 1% (5/487). Being bedridden was a common independent risk factor for colonization by all MDROs, although risk factors specific for each MDRO were identified. ESBL-producing Escherichia coli carriage was associated with the sequence type (ST) 131-H30 subclone, but other minor STs predominated in individual LTCF or in LTCFs located in the same city, suggesting a role for intrafacility or local transmission. Similarly, MRSA from LTCF residents belonged to the same spa types/ST clones (t008/ST8 and t032/ST22) commonly found in Italian acute-care hospitals, but infrequent spa types were recovered in individual LTCFs. The prevalent C. difficile PCR ribotypes were 356/607 and 018, both common in Italian acute-care hospitals. CONCLUSIONS: MDRO colonization is common among residents in Italian LTCFs.

Toxoplasma infection in individuals in central Italy: does a gender-linked risk exist?

An accurate estimate of the impact of toxoplasmosis on the population in Italy is not available. We performed a cross-sectional study on individuals living in Italy to assess: (1) differences in access to Toxo testing and in the prevalence of recent and past Toxoplasma gondii infection according to gender and age, and (2) the clinical impact of disease burden on the male patient subset. Reason for testing, condition of in- or outpatient and clinical data were analysed. Between-gender differences were observed in access to the test. Immunoglobulin M (IgM) prevalence was increased in males in the age range 5-34 years [odds ratio (OR) = 2.03, 95% confidence interval (CI) 1.18-3.49, p = 0.01), with a peak at 25-34 years. In females, it decreased in the age range 20-39 years (OR = 0.49, 95% CI 0.32-0.74, p = 0.0008). The attack rate of recent infection was twice as high for males than for females. Estimates pointed out 3.3 and 1.7 events in 1000 at-risk person-years in the male and female cohorts, respectively. Most IgM-positive subjects did not experience severe forms of toxoplasmosis, with 35% having lymphadenopathy. Chorioretinitis, systemic and neurological manifestations were also observed. Our findings suggest that the acute phase of toxoplasmosis is largely unapparent or clinically mild in this area. It is also possible that the disease burden for Toxoplasma infection in Italy is underestimated. Further study should focus on information acquisition and Toxo test access in hospital units for a better estimation of the real burden of mild and severe forms of the disease.

Seroprevalence for toxoplasmosis in individuals living in north west Tuscany: access to Toxo-test in central Italy.

This survey aimed to estimate the prevalence of anti-Toxoplasma IgG and IgM antibodies in people living in north west Tuscany (central Italy) and to investigate the adherence to antenatal screening programs and access to the Toxo-test as well. Sera from a large sample of individuals suspected to have acute infection or from pregnant women (10,352 subjects) aged between 1 day and over 70 years were analysed for both IgG and IgM anti-Toxoplasma antibodies using an immunoenzymatic method or a chemo-luminescent immunoassay. Overall, the seroprevalence of IgG antibodies was 21.4% (95% CI 20.62-22.20). A positive trend according to age was found, with low positivity observed in younger age groups. Among women of reproductive age the prevalence of IgG antibodies was 19.4% (95% CI 18.64-20.26). The overall IgM seroprevalence was 1.07% (95% CI 0.87-1.27). A low IgM prevalence was also observed in women of reproductive age (0.8%; 95% CI 0.65-1.03). Our study seems to indicate that primary prevention is widespread among women. However, an epidemiological surveillance system for toxoplasmosis should be implemented, to assess the risk of congenital toxoplasmosis and to determine the true burden of disease in adults.

The structure of Venus' middle atmosphere and ionosphere.

The atmosphere and ionosphere of Venus have been studied in the past by spacecraft with remote sensing or in situ techniques. These early missions, however, have left us with questions about, for example, the atmospheric structure in the transition region from the upper troposphere to the lower mesosphere (50-90 km) and the remarkably variable structure of the ionosphere. Observations become increasingly difficult within and below the global cloud deck (<50 km altitude), where strong absorption greatly limits the available investigative spectrum to a few infrared windows and the radio range. Here we report radio-sounding results from the first Venus Express Radio Science (VeRa) occultation season. We determine the fine structure in temperatures at upper cloud-deck altitudes, detect a distinct day-night temperature difference in the southern middle atmosphere, and track day-to-day changes in Venus' ionosphere.

Neonatal adrenal hemorrhage mimicking an acute scrotum.

Twenty-two cases of scrotal hematoma caused by neonatal adrenal hemorrhage are reported in the literature and unnecessary surgical exploration was performed in nine (41%), suspecting testicular torsion. In this paper, we present a newborn male with right adrenal gland hemorrhage causing right scrotal swelling and discoloration of groin managed conservatively.

Insulin glargine in patients with severe hepato-gastroenterology diseases and hyperglycemia receiving parenteral nutrition.

OBJECTIVE: The aim of this nonrandomized observational study is to verify and confirm whether it is possible to use insulin glargine (Lantus) subcutaneously in patients receiving parenteral nutrition (PN) and whether the analogue is capable of obtaining and maintaining good glycemic control without inducing hypoglycemia. MATERIALS AND METHODS: The sample is made up of 25 patients with severe hepato-gastroenterology diseases receiving parenteral nutrition, diagnosed diabetics and non-diabetics, who had previously been prescribed traditional insulin therapy. All were to be given subcutaneous insulin glargine at a dosage equal to the average of insulin/day administered in the preceding days spent receiving PN. RESULTS: Twenty-five consecutive patients, not stratified in any way, were judged eligible in the last six months of 2004 and first eight months of 2005. Four out of these 25 cannot be evaluated, either because (2/4) they did not begin or complete the treatment with Lantus or because the proper number of glycemic tests were not done (2/4); 21/25 patients, 84% of the sample with a mean age of 65.9 years (range 46-93 yr), finished the study and can be evaluated. The mean glycemic values after treatment with glargine were already better on the second day, and on the seventh day the difference was statistically significant. No hypoglycemias occurred which required medical intervention. CONCLUSIONS: This study confirms the possibility of using insulin glargine in patients receiving parenteral nutrition with hyperglycemia diagnosed diabetics or not diabetics.

Use of insulin glargine in patients with hyperglycaemia receiving artificial nutrition.

The purpose of this study was to verify whether it is possible to use insulin glargine (Lantus) subcutaneously in patients receiving artificial nutrition (AN) and if the analogue is capable of obtaining and maintaining good glycaemic control without inducing hypoglycaemia. The sample considers 25 patients receiving AN, diagnosed diabetics and non-diabetics, who had previously been prescribed traditional insulin therapy. All were to be given subcutaneous insulin glargine at a dosage equal to the average of insulin/day administered in the preceding days spent receiving AN. Twenty-five consecutive patients, not stratified in any way, were judged eligible in the last six months of 2004 and first two months of 2005. Four out of these 25 could not be evaluated, either because they did not begin or complete the treatment with Lantus (3/4) or because the proper number of glycaemic tests were not carried out (1/4); 21/25 patients, 84% of the sample with a mean age of 68.7 years (range 46-91 years), finished the study and could be evaluated. The mean glycaemic values after treatment with glargine were already better on the second day, and on the seventh day the difference was statistically significant. No hypoglycaemia requiring medical intervention occurred. This study confirms the possibility of using insulin glargine in patients receiving AN with hyperglycaemia regardless of the type of nutrition and whether or not the patient is diabetic.

Variations in the level of some trace elements in hair of participants in the Italian expeditions in Antarctica.

As a part the Italian National Programme of Research in Antarctica (PNRA) a monitoring study has been undertaken to quantify the concentrations of some selected trace elements in human hair of participants in the Antarctic expeditions. Such concentrations may vary as a consequence of the extreme environmental conditions and changes in lifestyle experienced by participants in the expeditions, as some evidence in previous investigations seems to suggest. The present study regards samples collected on the occasion of the 2002-2003 expedition to the Italian Base of Terra Nova Bay (now Mario Zucchelli Base), i.e., just before the expedition and about one month later. Seven essential elements were taken into account, namely, Ca, Cu, Cr, Fe, Mg, Mn and Mo. Determinations were performed by Inductively Coupled Plasma Atomic Emission Spectrometry (ICP-AES) and Dynamic Reaction Cell Inductively Coupled Plasma Mass Spectrometry (DRC-ICP-MS). Data obtained were statistically treated by using the non-parametric Friedman test. The concentrations of Ca, Cu and Mg were found to decrease (P < 0.05). The stress caused by the severe environmental conditions might well play a role in the observed decrease.

Clinical and genetic linkage analysis of a large Venezuelan kindred with Usher syndrome.

OBJECTIVE: To undertake a comprehensive investigation into the very high incidence of congenital deafness on the Macano peninsula of Margarita Island, Venezuela. METHODS: Numerous visits were made to the isolated island community over a 4-year-period. During these visits, it became apparent that a significant number of individuals complained of problems with hearing and vision. Socioeconomic assessments, family pedigrees and clinical histories were recorded on standard questionnaires. All individuals underwent thorough otolaryngologic and ophthalmologic examinations. Twenty milliliters of peripheral venous blood was obtained from each participant. A genome-wide linkage analysis study was performed. Polymorphic microsatellite markers were amplified by polymerase chain reaction and separated on polyacrylamide gels. An ABI 377XL sequencer was used to separate fragments and LOD scores were calculated by using published software. RESULTS: Twenty-four families were identified, comprising 329 individuals, age range 1-80 years, including 184 children. All families were categorized in the lower two (least affluent) socioeconomic categories. A high incidence of consanguinity was detected. Fifteen individuals (11 adults, 4 children) had profound congenital sensorineural hearing loss, vestibular areflexia and retinitis pigmentosa. A maximum LOD score of 6.76 (Linkage >3.0), between markers D11s4186 and D11s911, confirmed linkage to chromosome 11q13.5. The gene myosin VIIA (MYO7A) was confirmed in the interval. Clinical and genetic findings are consistent with a diagnosis of Usher syndrome 1B for those with hearing and vision problems. CONCLUSIONS: We report 15 Usher syndrome 1B individuals from a newly detected Latin American socio-demographic origin, with a very high prevalence of 76 per 100,000 population.

Initial experience with laparoscopic adrenal surgery in children: is endoscopic surgery recommended and safe for the treatment of adrenocortical neoplasms?

Through a review of 83 cases reported in literature, including our experience of two successful right laparoscopic adrenalectomies performed in a 3-year-old girl for androgen-secreting adenoma and in a 9-year-old male for pheochromocitoma, we have anaIyzed the indications, the techniques and the results of laparoscopic or retroperitoneoscopic adrenalectomy in children. Nineteen pheochromocytomas, 18 neuroblastomas, 15 adenomas, 12 ganglioneuromas, 9 hyperplasias, 2 carcinomas, 1 teratoma and 1 adrenogenital syndrome have been treated with excellent results (follow-up 1 month to 4 years) with no evidence of recurrence. The age range is from 1 month to 16 years with an equal female/male ratio. The tumor size range from 10 mm to 80 mm with approximately a mean of 40 mm. Sixty-three laparoscopies of which 54 lateral and 9 anterior have been performed versus 14 posterior retroperitoneoscopies. Regarding right adrenalectomy, 29 were laparoscopic and only 2 retroperitoneoscopic. With regards to left adrenalectomy, 23 were laparoscopic and 9 retroperitoneoscopic. Looking at bilateral lesions, these tumors were approached laparoscopically in 6 cases alternating both sides and 1 retroperitoneoscopically (this case was already operated on the other side). The operative time ranged from 25 minutes in newborn to 320 minutes in bilateral cases with an average of 120 minutes. Conversion rate is higher in retroperitoneoscopy (30%) (4 cases out of 13), than laparoscopy (6%) (4 cases out of 63). Specifically, for right retroperitoneoscopic adrenalectomy conversion rate was 100% (2 cases out 2), for left retroperitoneoscopic adrenalectomy 22.2% (2 cases out 9), for right laparoscopic adrenalectomy 6.8% (2 cases out of 29) and for left laparoscopic adrenalectomy 8.6% (2 cases out of 23). Only in 1 case were blood transfusions requested. Hospital stays ranged from 35 hours to 17 days with an average of 4 days. In conclusion the indications of endoscopic adrenalectomy in children are not different from those of traditional surgery and the well-known advantages of laparoscopic adrenal surgery should be applied to pediatric patients. It seems that there is no age and tumor size limits for a well-trained surgical team. Lateral transperitoneal approach is the most utilized with the child positioned in 90-degree flank decubitus. Laparoscopy is undoubtly preferred for right adrenalectomy (93% of cases), while for left adrenalectomy retroperitoneoscopy has been used in 39% of the cases. Considering the conversion rate and on the basis of our experience with adults, we recommend laparoscopic adrenalectomy for both right and left adrenal lesions, but we think that the pediatric surgeon should feel free to choose the approach in which he/she is more skilled. However, the best surgical result will be achieved if the pediatric and adult surgeon collaborate with their different experiences. Lastly, we suggest the use of new technological devices such as Ultracision Harmonic Scalpel which was a critical factor in our two successful right adrenalectomies.

[Esophageal atresia with distal tracheo-esophageal fistula. Evolution of the treatment in the period of 1955-2000 at the Anna Meyer Children's Hospital of Florence]. / Atresia dell'esofago con fistola esofago-tracheale distale. Evoluzione del trattamento nel periodo 1955-2000 nell'Ospedale Pediatrico Anna Meyer di Firenze.

BACKGROUND: Over the past decades the esophageal atresia (EA) has represented the greatest challenging malformation encountered by the pediatric surgeon. Since then, there have been considerable advancements in the treatment of EA. In this paper the experience at the "Anna Meyer Children's Hospital of Florence" in regards to the surgical treatment of the EA from 1955 to present day is reported, so that the analysis of the various medical and surgical choices followed by the authors and their predecessors in this long period, can be an important learning tool for the EA management. METHODS: From 1955 to 2000, 223 newborns affected by EA with tracheoesophageal fistula (TEF) have been operated on out of a total of 250 cases of EA. Our experience has been subdivided into periods on the basis of homogeneous medical and surgical treatment adopted in that determined time. We have analyzed particularly the data of the last period 1995-2000, where there has been a well standardized protocol of treatment from the medical, surgical and intensive care points of view. RESULTS: The mortality rate has decreased from 44.8 to 3.4% with a significant reduction (p<0.001) between the years 1979-1983 and 1984-2000, due to the introduction of a perioperative treatment in the newborn intensive care unit. Moreover, a significant correlation (p<0.05) has been shown between low birthweight and associated malformations, two risk factors that however do not negatively influence the results of the treatment in the last period 1995-2000. CONCLUSIONS: A full integration between the surgeon and neonatologist is necessary in order to guarantee a good result. The risk connected to EA is not as much the surgical procedure as the presence or absence of associate malformations that are undetected in the prenatal diagnosis. It is suggested that, in order to further reduce the mortality and morbidity rate after EA correction, the number of prenatal diagnoses should be increased.

Laparoscopic decapsulation of a large epidermoid splenic cyst in a child using the UltraCision LaparoSonic Coagulating Shears.

Splenic cysts are rare in pediatric surgery. Congenital epidermoid cysts are exceptional representing only 2.5% of all splenic cysts in childhood. Nowadays, considering the short- and long-term complications of splenectomy in children, the management of epidermoid cyst consists of partial splenectomy or decapsulation of the cystic wall. To our knowledge, the case reported in this article describes the first successful laparoscopic decapsulation of an epidermoid splenic cyst in an 10-year-old child using the UltraCision LaparoSonic Coagulating Shears (LCS). Follow-up at six months confirms no recurrence. Laparoscopic splenic decapsulation provides minimal access and small surgical trauma for treating the cyst while preserving splenic function. The use of UltraCision LCS makes the laparoscopy safely, expeditiously, with minimal blood loss and short hospital stay.

Validation of a food frequency questionnaire for nutritional calcium intake assessment in Italian women.

OBJECTIVE: To evaluate a food frequency questionnaire assessing calcium intake in women. DESIGN: : Estimates of calcium intake from the food frequency questionnaire were compared with those from 14 day records from 206 Caucasian women aged 25-75 y in Siena, Italy. SUBJECTS: Subjects were randomly recruited from the residents list of the city of Siena, Italy. Of the 250 initially recruited, 39 did not meet the inclusion criteria or failed to complete the diet record and five outliers were excluded before the statistical analysis on the basis that their diet record was unlikely to represent habitual intake. RESULTS: Mean dietary calcium intakes were 829+/-255 (s.d.) mg/day from the questionnaire and 818+/-260 (s.d.) mg/day from the diet record. The mean difference in intake by the two methods (-11.3+/-116.4 mg/day) did not differ significantly from zero. Specificity in classifying women consuming less than 800 mg/day calcium was 86.6%, and sensibility in classifying women consuming more than 800 mg/day calcium was 89.4%. CONCLUSIONS: The food frequency questionnaire could be used in epidemiological studies to assess calcium intake in young to elderly women. The specificity in identifying low calcium intake subjects makes it useful also as an educational tool in diet counselling and for prescribing calcium supplementation.

Heteropteris aphrodisiaca O. Machado : effects of extract BST 0298 on the oxidative stress of young and old rat brains.

The plant Heteropteris aphrodisiaca O. Machado is widely employed in Brazilian popular medicine as a stimulant, a tonic and a remedy for nervous debility. Several other plant products with similar therapeutic applications may have such effects based on the antioxidant content of polyphenols (gallates, cathecols, etc.) and flavonoids. The aim of this work was to investigate the effects of Heteropteris aphrodisiaca O. Machado extracts on the antioxidant defences and oxidative stress of young and old rat brains. Tests of lipoperoxidation in vitro, comparing H. aphrodisiaca extracts with butylhydroxytoluene (BHT), a classic antioxidant, showed an efficient inhibition of lipoperoxidation (Q(1/2) = 3.3 microg/mL). Chronic treatment of young and old rats with 50 mg plant extract/kg body weight resulted in a significant increase in superoxide dismutase (SOD) activities (40%) in the brain of old rats, but no changes in catalase and glutathione peroxidase (GPX). No significant variations in antioxidant enzyme activities were observed in the brains of young rats. However, significantly lower levels of lipoperoxidation (30%) were detected in the brain of young rats upon plant extract administration, suggesting that H. aphrodisiaca extracts reduced the oxidative stress to brain lipids.

Comparison of Mycobacterium tuberculosis susceptibility testing performed with BACTEC 460TB (Becton Dickinson) and MB/BacT (Organon Teknika) systems.

The recently introduced automated culture systems MB/BacT (Organon Teknika, Belgium) was compared with radiometric BACTEC 460TB (Becton Dickinson, USA) to test antimicrobial susceptibility of Mycobacterium tuberculosis to first line drugs. On 113 strains 97.5% agreement was obtained, with the difference being not significant. Concordance was practically complete for the most important drugs, isoniazid and rifampin. The two methods however significantly differed for the time needed to complete the test; in fact MB/BacT required on the average five days more than BACTEC 460TB. Despite the delay in the completion of the test, the excellent reliability along with the elimination of radioactivity and full automation make MB/BacT an attractive alternative for susceptibility testing of M. tuberculosis.

Bariatric surgery: early results with a multidisciplinary team.

BACKGROUND: Bariatric surgery depends on complete preoperative study of morbid obesity, in order to obtain the treatment of choice. A multidisciplinary group was founded in 1998 at the University of Siena. METHODS: During 1998, 16 patients, with median weight 121.8 +/- 31 kg and median body mass index (BMI) 43 +/- 6, underwent bariatric surgery. A multidisciplinary assessment was used in order to evaluate psychological status, food intake problems and patient compliance, and hemodynamic, respiratory, metabolic and arthritic functions. 13 patients were submitted to laparoscopic surgery: in 11 adjustable gastric banding was performed and 2 were submitted to a vertical gastroplasty plus adjustable gastric banding. Three patients were operated via traditional laparotomy, due to previous abdominal surgery in 2 cases (submitted to an adjustable gastric banding) and one woman was submitted to a bilio-intestinal bypass according to the Hallberg technique, for her psychiatric troubles and coexisting systolic hypertension and uncontrolled diabetes. Monthly follow-up for each patient continues after 6 months. RESULTS: No morbidity or mortality has occurred. The median weight loss at three months, was 19.5 kg. Two cases required injection of 1 ml more of fluid into the port, respectively at 4 and 9 months. Fifteen days after surgery, seven patients (46%) had vomiting episodes, due to rapid food intake. All patients have shown an improvement of their comorbidities after surgery. CONCLUSION: Early results via the multidisciplinary team and laparoscopic banding have been satisfactory thus far.