The efficacy of Italian guidelines in promoting oral health in children and adolescents.

AIM: The purpose of the study was to evaluate the efficacy of the guidelines on oral health published by the Italian Health Ministry. STUDY DESIGN: 1239 patients (582 girls and 657 boys) with a mean age of 4.46 (SD 2.81) years were evaluated before application of the guidelines by paediatricians (T0) for the presence of caries, gingivitis, diseases of the oral mucosa, and malocclusion. Only patients aged 6 months, 3 years, 6 years, and 9 years at T0 were taken into consideration. All patients were reevaluated after application of the ministerial guidelines (T1). METHODS: the study took place over a 3-year period. Enrolled patients referred to the outpatient clinics of three paediatricians of the Province of Genoa who strictly applied the ministerial guidelines. STATISTICS: Chi square test analysis was performed to evaluate a statistically significant decrease in the incidence of caries, gingivitis and diseases of the oral mucosa. RESULTS: After a 3-year follow-up collected data underlined a statistically significant decrease in the incidence of dental caries, gingivitis and oral mucosal diseases after implementation of the recommended ministerial guidelines. A statistically significant decrease of malocclusions was also evident in the elder patients (12 years old at T1). CONCLUSION: The accurate implementation of the Guidelines is supported. Only collaboration and increased synergy between paedodontists and paediatricians can fulfill the objectives which were the reason for publishing the "National guidelines for the promotion of oral health and the prevention of oral diseases in developmental age".

Preventive effects of oral probiotic on infantile colic: a prospective, randomised, blinded, controlled trial using Lactobacillus reuteri DSM 17938.

Infants were recruited in four centres in North-West Italy. 138 infants were assessed for eligibility, 113 ones underwent randomisation and 105 completed the study. Newborns aged less than 10 days of life, with gestational age between 37 and 42 weeks, birth weight from 2,500 to 4,300 g and normal physical examination were recruitable. Premature infants and infants affected by outcomes of perinatal hypoxia or necrotising enterocolitis have been excluded. Patients were randomly assigned to receive five drops containing Lactobacillus reuteri DSM 17938 (108 cfu) with 400 UI of vitamin D3 or only 400 UI of vitamin D3 daily. The primary endpoints concern the administration of pain relieving agents (cimetropium bromide at least three times per week or simethicone at least five times per week) from baseline to 12 weeks. Additional analyses were done on the percentage of infants that switched from an exclusive breastfeeding to a partial or exclusive formula feeding from baseline to 12 weeks. Data concerning the number of calls to the paediatricians and the number of visits at paediatricians' ambulatories due to infantile colic have been collected by paediatrician and analysed. Comparing the two groups, the relative risk was 0.04 (95% confidence interval (CI)=0.01-0.31) for cimetropium bromide, 0.24 (95% CI=0.14-0.41) for simethicone and 0.37 (95% CI=0.17-0.80) for the administration of infant formula, showing a protective action of L. reuteri. The treatment group showed a lower number of paediatric consultations related to episodes of infant colic than the control group (P<0.0001). L. reuteri DSM 17938 supplementation at the tested dosage could reduce parental discomfort due to infantile colic. The consumption of this probiotic is associated with a reduction of paediatric consultations for infantile colic, as well as use of pain relieving agents and of infant formula.

Virtual crossmatch in kidney transplantation.

BACKGROUND: The Luminex Single-Antigen Beads (LSA) assay allows an accurate detection and characterization of preexisting donor-specific antibodies (DSA) in kidney transplant candidates. But the ability of LSA to detect quite low levels of antibodies makes it hard to correctly predict crossmatch results in donor selection. In this study we retrospectively analyzed the accuracy of our virtual crossmatch (v-XM) protocol, which was used for selection of potential kidney transplant recipients, in predicting the results of actual crossmatch (a-XM) in cadaver-donor renal transplantation. We also investigated correlation between negative a-XM results and strength/specificity of preformed DSA. METHODS: The correlation between negative v-XMs and a-XMs performed in 2007-2012 at the Regional Transplant Center of the Lazio Region, Italy, was analyzed. In carrying out v-XM, the donor HLA molecules against which patients showed LSA-detected DSA with normalized mean fluorescence intensity (MFI)≥5,000 were considered to be "unacceptable DSA," and LSA-DSA showing MFI<5,000 were defined as "acceptable DSA." All cadaver donors had been typed for HLA-A, -B, -DR, and -DQB molecules by sequence-specific primer methods. On the basis of a negative v-XM, we performed 507 a-XMs between serum samples from 256 renal transplant candidates and T/B lymphocytes from 302 cadaver donors with the use of both complement-dependent cytotoxicity (CDC) and flow cytometry (FC) methods. RESULTS: The v-XM negative results showed good correlation with both CDC and FC a-XMs (97% and 90%, respectively). The sensitivity of v-XM was 100%; this high value was related to the lack of false-negative DSA results. The limited specificity with both techniques (CDC-XM, 74%; FC-XM, 79%) was due to the presence of "acceptable" and/or anti-DQA/DPB DSA in some patient sera used to perform the a-XMs. During the study period, 171 (67%) of the 256 sensitized patients received a kidney transplant: 30% of these had "acceptable DSA" and/or anti-DQA/DPB DSA. No antibody-mediated rejection due to preformed HLA-DSA was observed. CONCLUSIONS: Our v-XM protocol showed high sensitivity in predicting donor-recipient immunologic compatibility. The results of this study also demonstrated the importance of evaluating DSA strength for implementing v-XM results in the selection of kidney transplant recipients. Moreover, the finding of anti-DQA/DPB DSA, especially in serum samples that gave positive results with the use of both CDC and FC a-XMs, highlights the importance of defining all of the donor HLA molecules to perform an accurate v-XM.

Enzymatic hydrolysis of sugarcane bagasse pretreated with acid or alkali

The aim of this study was to evaluate the performance of enzymatic hydrolysis of acid or alkali pretreated sugarcane bagasse for the production of fermentable sugars. The first step consisted of selection of commercial enzymes presenting the highest cellulolytic activities. After selection of four enzymes: HPL, CL, P1 and P4, their performances were tested in the bagasse pretreated with acid and alkali. The sugar content of the hydrolysates was analyzed by anion exchange liquid chromatography. Data showed that the joint action of 0.5 percent acid pretreatment, 121ºC, 30 minutes and enzyme CL provides the best results, 67.25 g of hexose and 148.13g of pentose per kg of dry bagasse.

A biomonitoring study assessing the residual biological effects of pollution caused by the HAVEN wreck on marine organisms in the Ligurian Sea (Italy).

Residual biological effects of the 1991 HAVEN oil spill off the Ligurian (Arenzano) coast were assessed in this study. Samples of the fish species Boops boops, Mullus barbatus, and Uranoscupus scaber were collected from two polluted sites near the HAVEN wreck and from an uncontaminated area. In addition to this, mussels were caged along the coast affected by the HAVEN disaster. The physiological status of fish and mussels was assessed using a battery of stress and exposure biomarkers. The PAH content of mussel and fish tissues was also analyzed. Significant biological responses were observed in lysosomal membrane stability, neutral lipid and lipofuscin accumulation and micronucleus frequency for mussels caged at two sites close to the HAVEN wreck. Chemical analyses indicated, however, that these effects are not caused by aromatic hydrocarbons. For this reason, we suggest that the aftermath of the HAVEN disaster contributes very little to coastal ecosystem pollution. This was also confirmed by the few biological effects observed in fish specimens (Boops boops) collected from surface waters. Nevertheless, it is important to point out that benthic fish displayed a stress syndrome potentially caused by aromatic hydrocarbons released from the oil tanker, as witnessed by an enhanced EROD activity and increased lipofuscin and neutral lipid lysosomal contents.

Renal allograft immune response is influenced by patient and donor cytokine genotypes.

This study investigated the impact of specific cytokine genotypes on the incidence of acute rejection episodes (ARE), chronic graft dysfunction (CGD), and anti-HLA donor-specific antibody (DS-Ab) production in 86 renal transplant recipients and 70 cadaveric donors. A PCR-SSP method was performed for the analysis of polymorphisms in TNF-alpha, IL-6, TGF-beta, IL-10, and IFN-gamma cytokines. DS-Ab monitoring of sera was performed using a FCXM analysis. Observed cytokine frequencies for patients and donors were not significantly different from the expected frequencies under Hardy-Weinberg equilibrium conditions. The evaluation in recipients revealed a higher frequency of DS-Ab-positive patients among the TNF-alpha high (50.0% vs 25.7%), and for the IL-10 cytokine a greater incidence of ARE-positive patients (35.8% vs 18.2%) with the high + intermediate, compared with the low genotype. The combined effect of these 2 genotypes predisposed to DS-Abs (71.4% vs 25.3%; P = 0.02; odds ratio [OR] = 7.37). As for the TGF-beta1 cytokine, we observed a higher number of CGD-positive patients among high compared with intermediate producers (14.3% vs 0%; P = .050). The analysis of donors revealed a significantly lower incidence of ARE-positive patients among recipients whose donors were carriers of the high IL-6 G/G-genotype compared with the G/C+C/C-genotypes (16.7% vs 41.2%; P = .03), suggesting a protective effect of the G/G genotype on ARE and a predisposing role of donor (-174)allele C. In addition, we noted an association between the IFN-gamma low A/A-genotype and a higher incidence of ARE (42.1% vs 0%; P = .002) and DS-Ab production (47.4% vs 12.5%; P = .02) compared with high producers.

Optimization of emergency department management of infants with bronchiolitis.

Bronchiolitis is the most common lower respiratory tract infection in infants < 2 years of age; in the last decades both incidence and hospitalization rate had increased, thus increasing sanitary burden. From November 2006 to March 2007, an experimental protocol was followed in the Emergency Department at G. Gaslini Children's Hospital, Genoa, Italy, which attempted to optimise the management of patients with bronchiolitis and to reduce the overall hospitalization rate therefore admitting only those patients with severe illness. All clinical evaluations of the patients were obtained administering a score (Bronchiolitis Clinical Score - BCS), to quantify both initial severity of illness and response to treatment. All patient were at first treated with inhaled epinephrine, supplemented with or substituted by other drugs, if needed, according to clinical evolution. Moreover, strict admission and discharge criteria were defined, taking into consideration the BCS, response to treatment and the presence of risk factors for severe disease, attempting to increase the role of the Short Stay Unit (SSU). The outcome evaluated were the percentage of patients discharged, admitted and managed through the SSU respectively, the length of stay and the readmission rate after discharge; data collected were then compared to that regarding patients with bronchiolitis presented at the ED from November 2005 to March 2006. Our data showed an increasing of both discharged patients (37.5% vs 25.22%) and patients managed through the SSU (25.83% vs 19.57%) and a related decrease of hospitalization (36.67% vs 55.22%); no significative difference was observed regarding the readmission rate between the two populations. We also observed a statistically significant reduction of the length of stay in the study population (2.07 +/- 2.56 vs 2.84 +/- 3.25, p = 0.005). In conclusion, the protocol proposed showed to be useful in optimizing the ED management of the patient with bronchiolitis, being able to safely reduce both admission rate and lenght of stay.

Graft failure due to hemolytic uremic syndrome recurrence.

The hemolytic uremic syndrome (HUS) is a severe disease characterized by microangiopathic hemolytic anemia, thrombocytopenia, and acute renal failure. We herein report our experience with a 43-year-old female patient who underwent a second cadaveric kidney transplantation in February 2005, for adult-onset HUS. The first renal transplantation, which was performed in 1996, required removal after 3 weeks for probable recurrence of HUS. The immunosuppressive regimen for the second transplant included basiliximab, tacrolimus, mycophenolate mofetil, and steroids. On postoperative day (POD) 7, she received steroid treatment for an acute rejection episode with improved renal function. On POD 19 due to worsening renal function, a graft biopsy showed HUS recurrence, thus we instituted hemodialysis and then plasmapheresis treatments. At two months after transplantation, the patient continued under plasmapheresis treatment due to clinical evidence of HUS. On POD 80, cytomegalovirus infection was diagnosed and intravenous gancyclovir treatment started for 3 weeks. After 110 days from transplant, a deterioration in renal function was evident: the graft was swollen and painful with Doppler ultrasound showing patency of both the renal artery and vein but, low blood flow. After 2 weeks of hemodialysis, the patient underwent transplantectomy. In adult-onset HUS the recurrence rate reduces graft survival, particularly among patients undergoing second transplantation.

X-linked adrenal hypoplasia congenita and hypogonadotropic hypogonadism: report on new mutation of the DAX-1 gene in two siblings.

OBJECTIVE: Adrenal hypoplasia congenita (AHC) is a hereditary disorder that leads to adrenal insufficiency and hypogonadotropic hypogonadism (HHG) in childhood. The gene responsible for the X-linked form, DAX-1 (dosage-sensitive sex-reversal, AHC, on the X-chromosome, gene 1)/NR0B1, encodes for an unusual member of the nuclear receptor superfamily. Deletions and point mutations in the DAX-1 gene have been described in more than 70 AHC families. Inter- and intra-familial variability in the clinical presentation of AHC has been observed. Here we present the clinical and genetic data of two brothers affected by AHC. SUBJECTS AND METHODS: Clinical heterogeneity was observed in the two brothers: the first presented with adrenal insufficiency in early infancy, while the second required no substitution therapy until 4 yr of age. Interestingly, mineralcorticoid hormone deficiency preceded cortisol deficiency in both brothers. HHG was observed at pubertal age in both patients and required substitution therapy with gonadal steroids. RESULTS: Sequence analysis revealed a novel mutation in the DAX-1 gene in the two brothers and in their carrier mother. The mutation, a three nucleotide deletion, results in the loss of leucine 278 (del278L). A missense mutation affecting the same leucine (L278P) was previously shown to cause marked reduction of repressor function with respect to the wild type protein in transcription assays. CONCLUSIONS: Missense mutations or amino acid loss in the DAX-1 gene are very rare. Their identification and genotype-phenotype correlation are important for the characterization of protein function and for patient management.

Radiological research in Europe: a bibliometric study.

We performed a bibliometric search to evaluate number and scientific "weight" of papers written by European radiologists, as compared with colleagues from other countries, to measure the contribution of European researchers to radiology journals, and to correlate bibliometric parameters with some socio-economic factors of the different European nations. We considered all peer-reviewed articles published by radiologists in biomedical journals quoted by ISI over the 1995-2000 period. To identify authors as radiologists, the string "radiol" had to appear in the address of the corresponding author, and his country was considered as the country of origin of the paper. The definition of Europe included the 15 countries of the European Union, plus Norway and Switzerland. The scientific "weight" of the paper was assumed to be the impact factor of the journal of the publication in that given year. Then, we considered the annual indexes number of papers/population and number of papers/Gross Domestic Product (GDP) in each country. Data were retrieved from the Eurostat annual statistic reviews. From these bases, we obtained a comparison of the scientific production among European radiologists, those from the U.S. and those from the rest of the world. European radiological research is responsible of almost 40% of the world scientific production in our field, and Germany, UK and France are the leading publishers in Europe. An increase of the number of papers written by European radiologists was noted in the 1995-2000 period, whereas the production from the U.S. had a slight decrease. The mean concentration indexes papers/inhabitants and papers/GDP were significantly lower in Europe than in the U.S., even if some small European countries had higher values than the U.S. As a mean, European research received a lower impact factor than that from the U.S. The assessment of research output has progressively developed as an important issue for the scientific research community. Although not flawless, and often criticized for a variety of reasons, citation analysis is a commonly used technique in this field, is a frequent means to "weight" the scientific production of researchers and is one of the criteria used to assign research grants. Our study shows that European radiology is growing and its production is increasing over time, thus indicating strong commitment to research from European radiologists; however, European radiological research has not yet reached leadership in the literature, and mean indexes addressing the level of resources allocated to research are lower in Europe than in the U.S. This latter point has notable exceptions, but indicates inadequacy of funding, at least in some nations, and in Europe as a whole. The development of research programs within the framework of the European Union specifically aimed to radiology could lead to further advancement of our discipline.

Impact of donor-specific antibodies on chronic rejection occurrence and graft loss in renal transplantation: posttransplant analysis using flow cytometric techniques.

BACKGROUND: Improvements in immunosuppressive therapy have greatly reduced acute rejection (ARj) episodes, ensuring better short-term graft outcome, but have not modified long-term survival in renal transplantation. It is now well accepted that chronic rejection (CRj) can be determined by both immune and/or nonimmune mechanisms. The aim of this study was to evaluate the importance of the posttransplant humoral immune response towards mismatched HLA graft antigens in CRj occurrence and graft outcome. METHODS: Serum samples from 120 nonpresensitized renal transplant recipients were prospectively screened for 1 year after surgery by means of flow cytometry cross-match (FCXM) and FlowPRA beads (microbeads coated with purified HLA class I and class II antigens) assays. All transplants were followed-up for 2 years or until graft removal. RESULTS: FCXM monitoring identified donor-specific antibodies (DS-Abs) in 29 (24.2%) of 120 transplanted patients. Correlation with clinical data highlighted a higher incidence of ARj in DS-Abs-positive patients compared to negative patients (62% vs. 13%, P<0.00001). Furthermore, graft failure occurred more frequently among FCXM-positive patients than among negative patients (34% vs. 1%, P<0.00001). The deleterious effect of DS-Abs on graft function was confirmed by serum creatinine levels 2 years after transplantation. These were in fact higher in subjects producing DS-Abs than in subjects with only ARj (mean creatinine: 2.5+/-1.3 mg/dL vs.1.7+/-0.5 mg/dL, P=0.04). FlowPRA analysis of DS-Ab HLA specificity highlighted the presence of anti-HLA class I antibodies in 85% of FCXM-positive patients, who also presented with a higher incidence of HLA-B mismatches than FCXM-negative patients (1.23+/-0.66 vs. 0.92+/-0.59, P=0.02). CONCLUSIONS: Flow cytometric techniques are precious tools for investigating the activation of the humoral response against HLA antigens of the graft in renal transplantation. DS-Abs production has a worse impact on organ function and survival than ARj episodes. These findings represent further proof of the threat posed by DS-Abs on long-term graft function and draw attention to the need for a specific immunosuppressive therapy aimed at counteracting the different kinds of immune activation toward graft.

Posttransplant donor-specific antibody characterization and kidney graft survival.

This study was designed to investigate the clinical relevance of donor-specific antibodies (DS-Abs) and their influence on graft survival. Among 106 patients who underwent cadaveric kidney donor transplantation and were monitored by flow cytometry crossmatch (FCXM) during the 1st posttransplantation year, 25 (23.6%) resulted positive for DS-Ab production. During a 2-year follow up only 12 of the 81 FCXM-negative patients (14.8%) suffered rejection vs 17 of 25 FCXM-positive patients (68%; P = 0.00001). Correlating graft loss to DS-Ab production, 9 FCXM-positive patients lost the graft vs only 1 among the FCXM-negative patients. A worse graft function was evidenced in FCXM-positive subjects who had also suffered rejection episodes than in those which had acute rejection but did not produce DS-Abs. A high incidence of HLA-AB mismatches was found in FCXM-positive subjects which produced anti-class I antibodies. FCXM appears useful in estimating posttransplant alloimmune response. Moreover our findings confirm the harmful effects of anti-class I DS-Abs on long-term graft survival.

Incidence and nature of the most common rugby injuries sustained in Argentina (1991-1997).

OBJECTIVE: To study the incidence of the most commonly sustained injuries in Argentine rugby and analyse them according to type, position and age of the players, and phase and time of play. METHODS: A prospective registry of injuries was constructed in different provincial unions of Argentina. Data were collected during a whole weekend each year from 1991 to 1997. Chi2 with Yates correction test, contingency tables, odds ratios (OR), and 95% confidence intervals (CI) were calculated (Epi Info Version 6.04a). RESULTS: A total of 924 injuries were registered in 1296 rugby games, involving 38 933 players. The mean (SD) incidence per weekend was 2.4 (0.2)% (95% CI 2.22 to 2.53), and the number of injuries per season was 24,188. Overall, senior players suffered more injuries than those in younger divisions (OR = 1.53; 95% CI 1.34 to 1.76; p<0.0001). The most common type of injury was pulled muscles of the lower limbs (11.7%, p<0.0001). Overall, the knee was the most susceptible to injury (14.1%, p<0.0001). Senior players suffered more pulled muscles of the lower limbs (OR = 2.99; 95% CI 2.01 to 4.46; p<0.0001), ankle ligament distension (OR = 1.69; 95% CI 1.12 to 2.53; p = 0.01), knee trauma (OR = 1.69; 95% CI 1.06 to 2.68; p = 0.02), bleeding wounds on the face (OR = 3.86; 95% CI 2.24 to 6.70; p<0.0001), and knee ligament distension (OR = 2.14; 95% CI 1.16 to 3.96; p = 0.01). Younger players had a greater risk of suffering muscular or ligament injuries of the cervical column (OR = 3.0; 95% CI 1.05 to 10.08; p = 0.04). The forwards had a higher risk of injury (OR = 1.41; 95% CI 1.23 to 1.61; p<0.0001). The most commonly injured player was the flanker (15.5%, p<0.01), and the most common mechanism was in open play (33%). More injuries were sustained in the second half (OR = 1.17; 95% CI 1.03 to 1.34; p = 0.01). CONCLUSIONS: Injuries are the cause of significant morbidity among rugby players in Argentina. A more thorough investigation and a greater understanding of the mechanisms are crucial in order to update the rugby laws and reduce this high injury incidence.

Disabling injuries of the cervical spine in Argentine rugby over the last 20 years.

OBJECTIVE: To investigate the incidence and risk factors of disabling injuries to the cervical spine in rugby in Argentina. METHODS: A retrospective review of all cases reported to the Medical Committee of the Argentine Rugby Union (UAR) and Rugby Amistad Foundation was carried out including a follow up by phone. Cumulative binomial distribution, chi 2 test, Fisher test, and comparison of proportions were used to analyse relative incidence and risk of injury by position and by phase of play (Epi Info 6, Version 6.04a). RESULTS: Eighteen cases of disabling injury to the cervical spine were recorded from 1977 to 1997 (0.9 cases per year). The forwards (14 cases) were more prone to disabling injury of the cervical spine than the backs (four cases) (p = 0.03). Hookers (9/18) were at highest risk of injury (p < 0.01). The most frequent cervical injuries occurred at the 4th, 5th, and 6th vertebrae. Seventeen of the injuries occurred during match play. Set scrums were responsible for most of the injuries (11/18) but this was not statistically significant (p = 0.44). The mean age of the injured players was 22. Tetraplegia was initially found in all cases. Physical rehabilitation has been limited to the proximal muscles of the upper limbs, except for two cases of complete recovery. One death, on the seventh day after injury, was reported. CONCLUSIONS: The forwards suffered a higher number of injuries than the backs and this difference was statistically significant. The chance of injury for hookers was statistically higher than for the rest of the players and it was particularly linked to scrummaging. However, the number of injuries incurred in scrums was not statistically different from the number incurred in other phases of play.