Phosphorus sorption and availability in an andosol after a decade of organic or mineral fertilizer applications: Importance of pH and organic carbon modifications in soil as compared to phosphorus accumulation.

The effect of organic fertilizers on soil phosphorus (P) availability is usually mainly associated with the rate and forms of P applied, while they also alter the soil physical-chemical properties, able to change P availability. We aimed to highlight the impact of pH and organic C modifications in soil on the inorganic P (Pi) sorption capacity and availability as compared to the effect of P accumulation after mineral or organic fertilizers. We conducted a 10-years-old field experiment on an andosol and compared fields that had been amended with mineral or organic (dairy slurry and manure compost) fertilizers against a non-fertilized control. Water and Olsen extractions and Pi sorption experiments were realized on soils sampled after 6 and 10 years of trial. We also realized an artificial and ex situ alkalization of the control soil to isolate the effect of pH on Pi sorption capacity. Organic fertilizer application increased total P, pH, and organic C in soil. Pi-Olsen increased mainly with soil total P (r2 adj = 0.79), while Pi-water increased jointly with soil total P and pH (r2 adj = 0.85). The Pi sorption capacity decreased with organic fertilizer application. Artificial and ex situ alkalization of the control soil showed that Pi sorption capacity decreased with increasing pH. Our study demonstrated that, beyond the P fertilization rate, the increase in organic C content and even more so in pH induced by a decade of organic fertilizer applications in soil decreased the Pi sorption capacity and consequently increased Pi-water in soil.

Phosphorus-acquisition strategies of canola, wheat and barley in soil amended with sewage sludges.

Crops have different strategies to acquire poorly-available soil phosphorus (P) which are dependent on their architectural, morphological, and physiological root traits, but their capacity to enhance P acquisition varies with the type of fertilizer applied. The objective of this study was to examine how P-acquisition strategies of three main crops are affected by the application of sewage sludges, compared with a mineral P fertilizer. We carried out a 3-months greenhouse pot experiment and compared the response of P-acquisition traits among wheat, barley and canola in a soil amended with three sludges or a mineral P fertilizer. Results showed that the P-acquisition strategy differed among crops. Compared with canola, wheat and barley had a higher specific root length and a greater root carboxylate release and they acquired as much P from sludge as from mineral P. By contrast, canola shoot P content was greater with sludge than with mineral P. This was attributed to a higher root-released acid phosphatase activity which promoted the mineralization of sludge-derived P-organic. This study showed that contrasted P-acquisition strategies of crops allows increased use of renewable P resources by optimizing combinations of crop and the type of P fertilizer applied within the cropping system.

The combined use of platelet rich plasma and hyaluronic acid: prospective results for the treatment of knee osteoarthritis.

Osteoarthritis represents an important social economic burden with a high incidence worldwide. Conservative management of knee OA consists in several therapeutic options: pharmacologic therapy such as analgesics, non-steroid and steroid anti-inflammatory drugs, physical therapy, and injective therapy with hyaluronic acid (HA) and platelet-rich plasma injections (PRP). The aim of our study is to evaluate the effect of combined autologous PRP and HHA (Hybrid Hyaluronic Acid) viscosupplementation on clinical outcomes of patients with knee OA, by assessing the subjects before and after injective treatment. The study was conducted on 60 patients with an age between 40 and 70 years old affected by unilateral symptomatic knee osteoarthritis (stage II and III of Kellgren-Lawrence scale) nonresponsive to pharmacologic and rehab treatment. We divided the patients in two groups, and we treated the group A with injection of HHA and group B with HHA+PRP. Each patient received 3 injections at an interval of 1 week for 3 consecutive weeks. The patients were evaluated by the Knee Injury and Osteroartrhitis Outcome Score (KOOS) and Visual Analog Scale (VAS) at 3, 6 and 12 months after treatment. Statistical comparison between groups showed a significantly better result for the group B concerning the KOOS value, at 3 months and at 6 months. This difference, although clinically relevant, lost the statistical significance at 12 months. The VAS trend differently showed a significant difference at 3 and 12 months, while at 6 months the superiority of group B did not achieve statistical significance. Few studies investigated the effects of HA+PRP combined treatment for knee OA. Numerous studies demonstrated the efficacy of HA injection therapy in knee OA for a clinical point of view, reducing the pain and improving the quality of life. PRP preparations also improved functional outcome scores compared to hyaluronic acid and placebo in patients affected by knee OA. Based on our results we can conclude that the combined PRP and HHA treatment is not only a safe and efficacious procedure which can provide functional benefit but is also significantly better than HHA injective therapy alone, as demonstrated by the comparison within our cohort.

Behaviour of S-metolachlor and its oxanilic and ethanesulfonic acids metabolites under fresh vs. partially decomposed cover crop mulches: A laboratory study.

At the time of spring pre-emergent herbicide application, the soil surface in conservation agriculture is most of the time covered by cover crops (CC) mulches. The state of these mulches depends on their destruction date and on the selected species. Sorption and degradation of 14C-S-metolachlor on and within 8 decaying CC-covered (2 species × 4 initial decomposition state) soils corresponding to conservation agriculture were compared to its fate in bare soil (BS) corresponding to conventional agriculture. 14C-S-metolachlor and its metabolites distribution between mineralized, extractable and non-extractable (NER) fractions was determined at 5 dates during a 20 °C/84-d period. Herbicide mineralization was weak (<2%) for both CC and BS. Extractability of 14C in BS was intermediate between CC that were decomposed 28 or 56 days and 0 or 6 days before application. Degradates consisted in up to 43% of total radioactivity, with specificities according to the CC or soil compartment. NER formation was equivalent in BS and in the much decomposed CC-amended microcosms, and was stronger in less decomposed CC. S-metolachlor DT50 was 23-d in BS, and 9, 15, 39 and 25-d for CC ordered by increased decomposition state at the time of application. These results were attributed to the proportion of 14C intercepted by CC, and to higher levels of organic matter and microbial activity in less decomposed CC as compared with more decomposed ones. Then the state of decomposition level of CC residues determines the behaviour of SMOC (S-metolachlor) sprayed on the mulch in the conditions of conservation agriculture.

Intraoperative application Platelet rich fibrin, postoperative injections OF PRP or microfracture only for osteochondral lesions of the knee: a five-year retrospective evaluation.

Cartilage lesions are the most common cause of chronic knee pain. Micro-fracturing is reliable, effective, easy to perform and inexpensive. We propose a novel approach to cartilage lesions where microfractures are performed contextually to intra-operative or post-operative administration of platelet concentrates. We retrospectively evaluate 48 patients divided in 3 groups. Group 1: 15 patients underwent microfractures and intraoperative administration of PRF (PRF group); group 2: 16 microfractures and postoperative injections of PRP (PRP group); group 3: 17 patients with isolated microfractures (Microfractures group). Clinical scores (IKDC, VAS pain) were administered at 2 and 5 years postoperative and MRI was performed to evaluate the lesions of patients according to the MOCART criteria (2006). Patients treated with platelet concentrates achieved better clinical results compared to patients treated with microfracture only. The PRF group showed better results than the PRP group at 2 years, with loss of significance at 5 years. At MOCART score, PRF group obtained better results earlier than the other two groups.

Comparing hybrid hyaluronic acid with PRP in end career athletes with degenerative cartilage lesions of the knee.

Cartilage lesions are very common causes of chronic knee pain in athletes. Current treatment options consist in conservative strategies, such as viscosupplementation and platelet-rich plasma injections. This randomized controlled trial aims to investigate the effect of intra-articular Hybrid Hyaluronic Acid injections compared to PRP for the treatment of cartilage lesions among athletes at the end of their career. Since March 2015, 48 professional soccer players were randomized into two groups: 24 patients received 3 injections of HHA and 23 patients received 3 intra-articular injections of PRP. All patients achieved a statistically significant clinical improvement from preoperative to postoperative time in both groups. Patients in the HHA group showed a significant superiority compared to PRP group at 3 and 6 months. Intergroup differences decrease gradually until loss of significance at 12 months follow-up. Athletes with chronic degenerative cartilage lesions of the knee responded positively both to HHA and PRP until last follow up.

Candida-streptococcal mucosal biofilms display distinct structural and virulence characteristics depending on growth conditions and hyphal morphotypes.

Candida albicans and streptococci of the mitis group form communities in multiple oral sites, where moisture and nutrient availability can change spatially or temporally. This study evaluated structural and virulence characteristics of Candida-streptococcal biofilms formed on moist or semidry mucosal surfaces, and tested the effects of nutrient availability and hyphal morphotype on dual-species biofilms. Three-dimensional models of the oral mucosa formed by immortalized keratinocytes on a fibroblast-embedded collagenous matrix were used. Infections were carried out using Streptococcus oralis strain 34, in combination with a C. albicans wild-type strain, or pseudohyphal-forming mutant strains. Increased moisture promoted a homogeneous surface biofilm by C. albicans. Dual biofilms had a stratified structure, with streptococci growing in close contact with the mucosa and fungi growing on the bacterial surface. Under semidry conditions, Candida formed localized foci of dense growth, which promoted focal growth of streptococci in mixed biofilms. Candida biofilm biovolume was greater under moist conditions, albeit with minimal tissue invasion, compared with semidry conditions. Supplementing the infection medium with nutrients under semidry conditions intensified growth, biofilm biovolume and tissue invasion/damage, without changing biofilm structure. Under these conditions, the pseudohyphal mutants and S. oralis formed defective superficial biofilms, with most bacteria in contact with the epithelial surface, below a pseudohyphal mass, resembling biofilms growing in a moist environment. The presence of S. oralis promoted fungal invasion and tissue damage under all conditions. We conclude that moisture, nutrient availability, hyphal morphotype and the presence of commensal bacteria influence the architecture and virulence characteristics of mucosal fungal biofilms.

LGI1 microdeletion in autosomal dominant lateral temporal epilepsy.

OBJECTIVES: To characterize clinically and genetically a family with autosomal dominant lateral temporal epilepsy (ADLTE) negative to LGI1 exon sequencing test. METHODS: All participants were personally interviewed and underwent neurologic examination. Most affected subjects underwent EEG and neuroradiologic examinations (CT/MRI). Available family members were genotyped with the HumanOmni1-Quad v1.0 single nucleotide polymorphism (SNP) array beadchip and copy number variations (CNVs) were analyzed in each subject. LGI1 gene dosage was performed by real-time quantitative PCR (qPCR). RESULTS: The family had 8 affected members (2 deceased) over 3 generations. All of them showed GTC seizures, with focal onset in 6 and unknown onset in 2. Four patients had focal seizures with auditory features. EEG showed only minor sharp abnormalities in 3 patients and MRI was unremarkable in all the patients examined. Three family members presented major depression and anxiety symptoms. Routine LGI1 exon sequencing revealed no point mutation. High-density SNP array CNV analysis identified a genomic microdeletion about 81 kb in size encompassing the first 4 exons of LGI1 in all available affected members and in 2 nonaffected carriers, which was confirmed by qPCR analysis. CONCLUSIONS: This is the first microdeletion affecting LGI1 identified in ADLTE. Families with ADLTE in which no point mutations are revealed by direct exon sequencing should be screened for possible genomic deletion mutations by CNV analysis or other appropriate methods. Overall, CNV analysis of multiplex families may be useful for identifying microdeletions in novel disease genes.

Familial temporal lobe epilepsy with psychic auras associated with a novel LGI1 mutation.

BACKGROUND: Autosomal dominant lateral temporal epilepsy (ADLTE) is characterized by focal seizures with auditory features or aphasia. Mutations in the LGI1 gene have been reported in up to 50% of ADLTE pedigrees. We report a family with temporal lobe epilepsy characterized by psychic symptoms associated with a novel LGI1 mutation. METHODS: All participants were personally interviewed and underwent neurologic examination and video-EEG recordings. LGI1 exons were sequenced by standard methods. Mutant cDNA was transfected into human embryonic kidney 293 cells; both cell lysates and media were analyzed by Western blot. In silico modeling of the Lgi1 protein EPTP domain was carried out using the structure of WD repeat protein and manually refined. RESULTS: Three affected family members were ascertained, 2 of whom had temporal epilepsy with psychic symptoms (déjà vu, fear) but no auditory or aphasic phenomena, while the third had complex partial seizures without any aura. In all patients, we found a novel LGI1 mutation, Arg407Cys, which did not hamper protein secretion in vitro. Mapping of the mutation on a 3-dimensional protein model showed that this mutation does not induce large structural rearrangements but could destabilize interactions of Lgi1 with target proteins. CONCLUSIONS: The Arg407Cys is the first mutation with no effect on Lgi1 protein secretion. The uncommon, isolated psychic symptoms associated with it suggests that ADLTE encompasses a wider range of auras of temporal origin than hitherto reported.

Mucosal tissue invasion by Candida albicans is associated with E-cadherin degradation, mediated by transcription factor Rim101p and protease Sap5p.

The ability of Candida albicans to invade mucosal tissues is a major virulence determinant of this organism; however, the mechanism of invasion is not understood in detail. Proteolytic breakdown of E-cadherin, the major protein in epithelial cell junctions, has been proposed as a mechanism of invasion of certain bacteria in the oral mucosa. The objectives of this study were (i) to assess whether C. albicans degrades E-cadherin expressed by oral epithelial cells in vitro; (ii) to compare the abilities of strains with different invasive potentials to degrade this protein; and (iii) to investigate fungal virulence factors responsible for E-cadherin degradation. We found that while E-cadherin gene expression was not altered, E-cadherin was proteolytically degraded during the interaction of oral epithelial cells with C. albicans. Moreover, C. albicans-mediated degradation of E-cadherin was completely inhibited in the presence of protease inhibitors. Using a three-dimensional model of the human oral mucosa, we found that E-cadherin was degraded in localized areas of tissue invasion by C. albicans. An invasion-deficient rim101-/rim101- strain was deficient in degradation of E-cadherin, and this finding suggested that proteases may depend on Rim101p for expression. Indeed, reverse transcription-PCR data indicated that expression of the SAP4, SAP5, and SAP6 genes is severely reduced in the rim101-/rim101- mutant. These SAP genes are functional Rim101p targets, because engineered expression of SAP5 in the rim101-/rim101- strain restored E-cadherin degradation and invasion in the mucosal model. Our data support the hypothesis that there is a mechanism by which C. albicans invades mucosal tissues by promoting the proteolytic degradation of E-cadherin in epithelial adherens junctions.

Genetic analysis of the LGI/Epitempin gene family in sporadic and familial lateral temporal lobe epilepsy.

Mutations in the LGI1/Epitempin gene cause autosomal dominant lateral temporal lobe epilepsy (ADLTE), a partial epilepsy characterized by the presence of auditory seizures. However, not all the pedigrees with a phenotype consistent with ADLTE show mutations in LGI1/Epitempin, or evidence for linkage to the 10q24 locus. Other authors as well as ourselves have found an internal repeat (EPTP, pfam# PF03736) that allowed the identification of three other genes sharing a sequence and structural similarity with LGI1/Epitempin. In this work, we present the sequencing of these genes in a set of ADLTE families without mutations in both LGI1/Epitempin and sporadic cases. No analyzed polymorphisms modified susceptibility in either the familial or sporadic forms of this partial epilepsy.

Increased expression of LGI1 gene triggers growth inhibition and apoptosis of neuroblastoma cells.

The LGI1 gene has been implicated in the malignant progression of glioblastoma and it has also been genetically linked to a form of partial epilepsy (ADLTE). In this study, we investigated the relevance of LGI1 expression for neuroblastoma cells. The analysis of two cell lines (SH-SY5Y and SK-N-BE) revealed unpredictably low levels of LGI1 and stable cell transfection with LGI1 cDNA yielded moderate increases of LGI1 expression. Neuroblastoma cell clones exhibited impaired cell growth and survival ability in relation to LGI1 levels. The process of growth inhibition could be discerned under experimental conditions of low cell density, since conditions of elevated cell density, which enhance the requirement for survival stimuli, resulted in massive cellular death. At high cell density, spontaneous apoptosis of LGI1 cells was clearly shown by the release of cytochrome c and apoptosis inducing factor (AIF) from mitochondria and by phosphatydil serine exposure and nuclear fragmentation. Activation of apoptotic effectors caspase-3/7 also occurred, however, the broad caspase inhibitor Z-VAD-FMK substantially failed to block cell death. Thus the possibility that LGI1-triggered apoptosis may involve initiator caspases linked to activation of death receptors, appears unlikely. The decreased ratio of Bcl-2 to Bax suggests that apoptosis is initiated by the intrinsic mitochondrial pathway through the release of caspase-dependent and -independent apoptogenic molecules. This study provides the first evidence that LGI1 controls neuronal cell survival, suggesting its role in the development of the nervous system in relation to the pathogenesis of neuroblastoma and ADLTE.

Idiopathic partial epilepsy with auditory features (IPEAF): a clinical and genetic study of 53 sporadic cases.

The purpose of our study was to describe the clinical characteristics of sporadic (S) cases of partial epilepsy with auditory features (PEAF) and pinpoint clinical, prognostic and genetic differences with respect to previously reported familial (F) cases of autosomal dominant partial epilepsy with auditory features (ADPEAF). We analysed 53 patients (24 females and 29 males) with PEAF diagnosed according to the following criteria: partial epilepsy with auditory symptoms, negative family history for epilepsy and absence of cerebral lesions on NMR study. All patients underwent a full clinical, neuroradiological and neurophysiological examination. Forty patients were screened for mutations in LGI1/epitempin, which is involved in ADPEAF. Age at onset ranged from 6 to 39 years (average 19 years). Secondarily generalized seizures were the most common type of seizures at onset (79%). Auditory auras occurred either in isolation (53%) or associated with visual, psychic or aphasic symptoms. Low seizure frequency at onset and good drug responsiveness were common, with 51% of patients seizure-free. Seizures tended to recur after drug withdrawal. Clinically, no major differences were found between S and F patients with respect to age at onset, seizure frequency and response to therapy. Analysis of LGI1/epitempin exons failed to disclose mutations. Our data support the existence of a peculiar form of non-lesional temporal lobe epilepsy closely related to ADPEAF but without a positive family history. This syndrome, here named IPEAF, has a benign course in the majority of patients and could be diagnosed by the presence of auditory aura. Although LGI1 mutations have been excluded, genetic factors may play an aetiopathogenetic role in at least some of these S cases.

[Prevalence of glycopeptide resistant enterococci in hospitalized patients and in the community in Calabria (Italy)]. / Prevalenza di portatori ospedalieri e comunitari di enterococchi resistenti ai glicopeptidi.

The study evaluated prevalence of glycopeptide (vancomycin and teicoplanin) resistant enterococci (VRE) in hospitalized patients and in the community in Calabria (Italy). Rectal swabs were collected from hospitalized and community subjects. Information was gathered on demographics, ward and duration of hospital stay and antibiotics administered; in community subjects information on previous hospitalizations and antibiotics use in the previous year were also collected. Rectal swabs were plated on Enterococcosel agar alone and with addition of vancomycin (6 mg/l) and incubated aerobically at 37 degrees for 24-48 hours. Typical colonies were tested for Gram stain and catalase production. Enterococci were identified to the species level by biochemical tests. Antimicrobial susceptibility test and E-test were performed by the disk diffusion method to evaluate vancomycin and teicoplanin susceptibility. t-test and chi square test were performed to evaluate difference in means and proportions, respectively. Samples were collected from 211 subjects; 132 (62.6%) from community and 79 (37.4%) from hospitals. Enterococci were isolated from 145 subjects (68.7%), 72.7% and 62% in community and in hospital, respectively. The most frequent isolates were E. faecium (30.4%), followed by E. faecalis and E. durans (25.5%), and, finally, by Leuconostoc (18.6%). Only one E. faecium strain was resistant to both vancomycin and teicoplanin, isolated from a non-hospitalized 45 years old woman (1.3%). The results did not show emergence of VRE in our area, although surveillance to confirm these findings is needed, considering the substantial spread of antibiotics in hospitals and in the community.

Healthcare personnel and hand decontamination in intensive care units: knowledge, attitudes, and behaviour in Italy.

The purpose of this study was to evaluate knowledge, attitudes, and behaviour regarding hand decontamination in personnel of intensive care units (ICUs) in Italy. All ICU physicians and nurses in 19 and five randomly selected hospitals in Campania and Calabria (Italy) were mailed a questionnaire focusing on demographics and practice characteristics; knowledge about prevention of hospital acquired infection; attitudes and behaviour with respect to hand decontamination; and use of gloves. A total of 413 questionnaires were returned giving a response rate of 66.6%. Overall, 53.2% agreed with the correct responses on knowledge related to infection control, and this knowledge was significantly higher in neonatal and medicine-surgery wards and in larger ICUs. A positive attitude was reported by the large majority who agreed that hand decontamination reduces the risk of infection in patients (96.8%) and personnel (86.2%), and the positive attitude was significantly higher among older and female personnel and in those with a higher level of knowledge. Only 60% always decontaminate hands at the start of a shift, and 72.5% before and after a patient contact. Higher compliance is reported for invasive manoeuvres, such as urinary catheters (96.5%) and intravenous lines (77.1%). Routine hand decontamination between each patient was significantly higher in females, and in neonatal and medicine-surgery ICUs. Our results suggest that interventions should not only be focused on predisposing factors (knowledge), but also on enabling (facilitating) and reinforcing (gratifying) factors.

Identification and characterization of a novel human brain-specific gene, homologous to S. scrofa tmp83.5, in the chromosome 10q24 critical region for temporal lobe epilepsy and spastic paraplegia.

We describe the structure, genomic organization, and some transcription features of a human brain-specific gene previously localized to the genomic region involved in temporal lobe epilepsy and spastic paraplegia on chromosome 10q24. The gene, which consists of six exons disseminated over 16 kb of genomic DNA, is highly homologous to the porcine tmp83.5 gene and encodes a putative transmembrane protein of 141 amino acids. Unlike its porcine homolog, from which two mRNAs with different 5'-sequences are transcribed, the human gene apparently encodes three mRNA species with 3'-untranslated regions of different sizes. Mutation analysis of its coding sequence in families affected with temporal lobe epilepsy or spastic paraplegia linked to 10q24 do not support the involvement of this gene in either diseases.

Antibodies to the E2 protein of GB virus C/hepatitis G virus: prevalence and risk factors in different populations in Italy.

BACKGROUND: Many aspects of the epidemiology of GB virus C/hepatitis G virus (GBV-C) infection have not been fully elucidated. The purpose of this study was to assess the prevalence of GBV-C antibodies and risk factors in different subjects living in Italy. MATERIALS AND METHODS: A total of 1,005 sera were tested for the presence of antibodies to the of GBV-C E2 protein using a recently developed enzyme immunoassay. RESULTS: A high prevalence of GBV-C antibodies was found in healthy blood donors (12.6%). Hemodialysis patients and drug users showed higher rates of GBV-C seropositivity (22% and 39%, respectively). Immigrants from sub-Saharan Africa and South Asia had anti-GBV-C prevalence comparable to that found in Italian blood donors, whereas higher and lower rates were detected in immigrants from Latin America and the Caribbean (19.5%) and from Mediterranean Africa (5.6%). CONCLUSION: GBV-C infections are widespread in the general population in Italy and particularly common in risk groups. The different prevalence of GBV-C antibodies detected in third world immigrants is likely to reflect the prevalence in the countries of origin. However, the observation that the length of residency in Italy is a significant risk factor may suggest that at least some GBV-C infections are contracted in Italy.

Cigarette smoking: knowledge, attitudes and behaviour in an adult population in Italy.

The study explores knowledge, attitudes, and behaviour regarding cigarette smoking and related factors in an adult population. A total of 935 parents of children attending the eighth class of ten randomly selected primary schools in Catanzaro (Italy) received a questionnaire consisting of questions on demographic and socioeconomic characteristics, knowledge, behaviour and attitudes about cigarette smoking. Six hundred and sixty-nine parents returned the questionnaire, with a response rate of 71.5%. Knowledge of risk associated to smoking was significantly higher in more educated subjects and in past smokers compared to current. Current and past smokers were respectively 39.6% and 17.2%, and current smokers were younger, not married, less prone to consider smoking as a major risk for their health and more likely to live with other smokers compared to past and never smokers. Females of higher education were more likely to be current smokers, whereas male current smokers were more likely to be less educated compared to past or never smokers. The results strongly recommend the provision of accurate information about the health consequences related to smoking, with a more intensive involvement of health care providers, particularly targeted to women and younger age groups.