Celocentesis for early prenatal diagnosis of hemoglobinopathy.

OBJECTIVE: Celocentesis is an invasive technique that can provide prenatal diagnosis of single-gene disorders, from as early as 7 weeks' gestation. The objective of this study was to examine the safety of celocentesis. METHODS: In this prospective study, celocentesis was performed for prenatal diagnosis of hemoglobinopathy in 402 singleton pregnancies in which both parents were carriers of ß-thalassemia or sickle cell disease trait. We assessed procedure-related maternal discomfort or pain, success of sampling and obtaining results, pregnancy outcome and postnatal follow-up. RESULTS: First, celocentesis was carried out at a median gestational age of 8.6 (range, 6.9-9.9) weeks and celomic fluid was successfully aspirated in 99.8% of cases. Second, 67% of women had no or only mild discomfort, 18% had moderate discomfort, 12% had mild-to-moderate pain and 3% had severe pain. Third, prenatal diagnosis from analysis of the celomic fluid was successful in 93.8% cases, and in the last 121 cases, it was always successful. Fourth, in all cases of successful sampling and analysis of celomic fluid, the diagnosis was concordant with results obtained from additional prenatal or postnatal testing. Fifth, in addition to diagnosis of hemoglobinopathy, quantitative fluorescence polymerase chain reaction analysis, which was performed to evaluate maternal contamination using several markers for chromosomes X, Y, 21, 18 and 13, led to the accurate diagnosis of chromosomal aneuploidy. Sixth, in all cases of an affected fetus diagnosed by celocentesis in which the parents chose termination of pregnancy, this was carried out < 10 weeks' gestation. Seventh, in 97.1% (298/307) of the continuing pregnancies there was live birth, in seven (2.3%) there was miscarriage and in two (0.7%) there was loss to follow-up. Eighth, fetal abnormalities were diagnosed in three (1%) cases, including unilateral transverse amputation of the forearm, unilateral moderate hydronephrosis and small-bowel duplication. All neonates were examined by a pediatrician and were found to be phenotypically normal, except for the three cases with a prenatally diagnosed defect. CONCLUSIONS: Celocentesis can be used for early prenatal diagnosis of genetic abnormalities, and the procedure-related risk of pregnancy complications appears to be low. Copyright © 2020 ISUOG. Published by John Wiley & Sons Ltd.

Influence of salinity and sedimentation on Vibrio infection of the Hawaiian coral Montipora capitata.

Environmental cofactors alter host-pathogen interactions and influence disease dynamics by impairing host resistance and/or increasing pathogen virulence. Terrestrial runoff is recognized as a major threat to coral reef health. However, the direct links between runoff and coral disease are not clear. Montipora white syndrome (MWS) is a coral disease that occurs in the Hawaiian archipelago, can be caused by various bacterial pathogens, including Vibrio species, and is linked to conditions associated with heavy rainfall and runoff. The objective of this study was to determine whether a short-term hyposalinity stress (20 ppt for 24 h) or sedimentation stress (1000 g m-2 d-1) would influence bacterial infection of the coral Montipora capitata. Hyposalinity increased M. capitata susceptibility to infection by 2 MWS pathogens, Vibrio coralliilyticus strain OCN008 and Vibrio owensii strain OCN002. Specifically, hyposalinity allowed OCN008 to infect at lower doses (106 CFU ml-1 compared with 108 CFU ml-1) and reduced the amount of time before onset of OCN002 infection at high doses (108 CFU ml-1). In contrast, short-term sedimentation stress did not affect M. capitata infection by either of these 2 pathogens. Although several studies have found a correlation between runoff and increased coral disease prevalence in field studies, this is the first study to show that one aspect of runoff (reduced salinity) enhances bacterial infection of coral using manipulative experiments.

Impact of a rectal and bladder preparation protocol on prostate cancer outcome in patients treated with external beam radiotherapy.

PURPOSE AND OBJECTIVE: To test the hypothesis that a rectal and bladder preparation protocol is associated with an increase in prostate cancer specific survival (PCSS), clinical disease free survival (CDFS) and biochemical disease free survival (BDFS). PATIENTS AND METHODS: From 1999 to 2012, 1080 prostate cancer (PCa) patients were treated with three-dimensional conformal radiotherapy (3DCRT). Of these patients, 761 were treated with an empty rectum and comfortably full bladder (RBP) preparation protocol, while for 319 patients no rectal/bladder preparation (NRBP) protocol was adopted. RESULTS: Compared with NRBP patients, patients with RBP had significantly higher BDFS (64% vs 48% at 10 years, respectively), CDFS (81% vs 70.5% at 10 years, respectively) and PCSS (95% vs 88% at 10 years, respectively) (log-rank test p < 0.001). Multivariate analysis (MVA) indicated for all treated patients and intermediate high-risk patients that the Gleason score (GS) and the rectal and bladder preparation were the most important prognostic factors for PCSS, CDFS and BDFS. With regard to high- and very high-risk patients, GS, RBP, prostate cancer staging and RT dose were predictors of PCSS, CDFS and BDFS in univariate analysis (UVA). CONCLUSION: We found strong evidence that rectal and bladder preparation significantly decreases biochemical and clinical failures and the probability of death from PCa in patients treated without daily image-guided prostate localization, presumably since patients with RBP are able to maintain a reproducibly empty rectum and comfortably full bladder across the whole treatment compared with NRPB patients.

Incidence of haemoglobinopathies in Sicily: the impact of screening and prenatal diagnosis.

BACKGROUND: Haemoglobinopathies are a major public health problem in Sicily: it was estimated a frequency of 1/245 couples are at risk of haemoglobinopathies. This paper reviews legislative actions, prevention activities, carrier screening, genetic counselling, foetal sampling and laboratory methodology analysis evolution reporting the results of 30 years of prevention actions to assess the efficiency of our preventative programme in the control of haemoglobinopathies in Sicily. METHODS: This programme consisted principally of five phases: legislative actions, public awareness campaign, carrier screening, genetic counselling and prenatal diagnosis. RESULTS: These programmes have been very effective, which we can see from a greater public awareness of thalassaemia and its prevention in the target population furthermore by a marked decline in the incidence of thalassaemia major and sickle cell anaemia from 1 in 245 live births in the absence of prevention to 1 in 2000, with a reduction in about 85%. The residual cases were because of a conscious choice by expecting parents in relation to improved life expectancy as well as improved quality of life of the affected patients. CONCLUSION: The study suggests that public health authorities should act and invest in a similar programme for prevention of thalassaemia, as well as in relation to the increased survival of patients and the consequent organ complications.

Impact of COVID-19 lockdown on adolescents in treatment for obesity.

OBJECTIVE: The aim of this study was to evaluate the impact of coronavirus disease 2019 (COVID-19) lockdown on body mass index (BMI) z-score of adolescents treated for obesity, and to assess the correlation between BMI z-score changes and lifestyle variables. METHODS: This cross-sectional study recruited 66 participants aged 11-18 years with overweight or obesity who attended our obesity clinics before and after the lockdown. We assessed demographic and anthropometric characteristics as well as lifestyle variables. RESULTS: The mean BMI z-score of the participants was stable during this period (p = 0.233). Balanced diet, snacking, and sweet beverage consumption improved in about half of the participants. Having at least one parent at home was associated with a significantly more balanced diet (p = 0.008) and an increase in family activities (p = 0.015). Physical activities decreased (51.5%) and screen time increased for most of the adolescents (86.4%). The BMI z-score decreased significantly when three or more lifestyle habits improved (p<0.001). CONCLUSIONS: This study showed that BMI z-scores were stable for a majority of adolescents treated for obesity during the COVID19 lockdown, along with positive and negative changes on weight-related lifestyle habits. This extraordinary period may have created opportunities for lifestyle modification and has emphasized the importance of family support in the management of obesity in adolescents.

Pretreatment patient-specific IMRT quality assurance: a correlation study between gamma index and patient clinical dose volume histogram.

PURPOSE: The aim of this work is to investigate the predictive power of a common conventional intensity modulated radiation therapy (IMRT) quality assurance (QA) performance metric, the gamma passing rate (%GP), through the analysis of the sensitivity and of the correlation between %GP and different dose discrepancies between planned dose-volume histogram (DVH) and perturbed DVH. The perturbed DVH is calculated by using a dedicated software, 3DVH (Sun Nuclear Corporation, Melbourne, FL), which is able to modify the dose distribution calculated by the treatment planning system (TPS) according to the dose discrepancies detected with planar measurements in order to predict the delivered 3D dose distribution in the patient. METHODS: Twenty-seven high-risk prostate cancer (PP) patients and 15 head and neck (HN) cancer patients, treated with IMRT technique, were analyzed. Pretreatment verifications were performed for all patients' plans by acquiring planar dose distributions of each treatment field with 2D-diode array. Measured dose distributions were compared to the calculated ones using the gamma index (GI) method applying both global (Van Dyk) and local normalization, and %GP were generated for each pair of planar doses using the following acceptance criteria: 1%∕1, 2%∕2, and 3%∕3 mm. Planar dose distributions acquired during pretreatment verifications, together with patient's DICOM RT plan, RT structure set, and RT dose files from TPS were loaded into the 3DVH software. Percentage dose differences (%DE) between DVHs, obtained by TPS and by 3DVH, were calculated; statistical correlation between %DE and %GP was studied by using Pearson's correlation coefficient (r). This analysis was performed, for each patient, on planning target volumes and on some typical organs at risk of the prostatic and head and neck anatomical district. The sensitivity was calculated to correctly identify the pretreatment plans with high dose errors and to quantify the incidence of false negatives, on varying the gamma index method. RESULTS: Analysis of %DE vs %GP showed that there were only weak correlations (Pearson's r-values < 0.8). The results also showed numerous instances of false negatives (cases where high IMRT QA passing rates did not imply good agreement in anatomy dose metrics) and the reverse, mainly for the 3%∕3 mm global gamma passing rate. CONCLUSIONS: The lack of correlation between conventional IMRT QA performance metrics gamma passing rates and dose errors in DVHs values and the low sensitivity of 3%∕3 mm global gamma method show that the most common published acceptance criteria have disputable predictive power for per-patient IMRT QA.

The efficient physiological strategy of a novel tomato genotype to adapt to chronic combined water and heat stress.

Climate change is increasing the frequency of high temperature shocks and water shortages, pointing to the need to develop novel tolerant varieties and to understand the mechanisms employed to withstand combined abiotic stresses. Two tomato genotypes, a heat-tolerant Solanum lycopersicum accession (LA3120) and a novel genotype (E42), previously selected as a stable yielding genotype under high temperatures, were exposed to single and combined water and heat stress. Plant functional traits, pollen viability and physiological (leaf gas exchange and chlorophyll a fluorescence emission measurements) and biochemical (antioxidant content and antioxidant enzyme activity) measurements were carried out. A Reduced Representation Sequencing approach allowed exploration of the genetic variability of both genotypes to identify candidate genes that could regulate stress responses. Both abiotic stresses had a severe impact on plant growth parameters and on the reproductive phase of development. Growth parameters and leaf gas exchange measurements revealed that the two genotypes used different physiological strategies to overcome individual and combined stresses, with E42 having a more efficient capacity to utilize the limiting water resources. Activation of antioxidant defence mechanisms seemed to be critical for both genotypes to counteract combined abiotic stresses. Candidate genes were identified that could explain the different physiological responses to stress observed in E42 compared with LA3120. Results here obtained have shown how new tomato genetic resources can be a valuable source of traits for adaptation to combined abiotic stresses and should be used in breeding programmes to improve stress tolerance in commercial varieties.

Decreased bone turnover in children and adolescents with well controlled type 1 diabetes.

AIM: Osteoporosis is a common long-term complication of type 1 diabetes (T1DM). We aimed to determine whether bone mineral density (BMD) and turnover are already altered during childhood. PATIENTS AND METHODS: We recruited 27 T1DM children and 32 controls (age 10.5 +/- 2.5 yr.) and measured BMD (dual-energy x-ray absorptiometry); bone biomarkers levels (osteocalcin: OC; procollagen type 1 propeptides amino-terminal: PINP; crosslinking telopeptides of type 1 collagen C-terminal: CTX), glycated hemoglobin (HbA1c), dietary intake and physical activity. RESULTS: Patients with T1DM had lower levels of OC (70.3 +/- 3.3 vs 105.3 +/- 6.8), PINP (556.4 +/- 47.6 vs 716.3 +/- 53.8), CTX(0.97 +/- 0.07 vs 1.20 +/- 0.08), physical activity, and calcium intake. Biomarkers were negatively correlated with HbA1c. Though, BMD was similar among groups and not related to HbA1c, disease duration, physical activity or dietary intakes. CONCLUSIONS: Bone turnover is altered in T1DM children, whereas BMD remains normal during growth. Physical activity and optimal calcium intakes may improve bone metabolism and delay osteoporosis.

Quality indicators for hyperthermia treatment: Italian survey analysis.

AIM: Nowadays, no Quality Indicators (QI) have been proposed for Hyperthermia treatments. Starting from radiotherapy experience, the aim of this work is to adapt radiotherapy indicators to Hyperthermia and to propose a new specific set of QI in Hyperthermia field. MATERIAL AND METHODS: At first, radiotherapy quality indicators published in literature have been adapted to hyperthermia setting. Moreover, new specific indicators for the treatment of hyperthermia have been defined. To obtain the standard reference values of quality indicators, a questionnaire was sent to 7 Italian hyperthermia Institutes with a list of questions on physical and clinical hyperthermia treatment in order to highlight the different therapeutic approaches. RESULTS: Three structure, five process and two outcome QI were selected. It has been possible to adapt seven indicators from radiotherapy, while three indicators have been defined as new specific indicators for hyperthermia. Average values used as standard reference values have been obtained and proposed. CONCLUSION: The survey performed on 7 Italian centres allowed to derive the standard reference value for each indicator. The proposed indicators are available to be investigated and applied by a larger number of Institutes in which hyperthermia treatment is performed in order to monitor the operational procedures and to confirm or modify the reference standard value derived for each indicator.

Local anaesthetic vs. general anaesthetic for inguinal hernia repair: systematic review and meta-analysis.

OBJECTIVE: Whilst local anaesthesia (LA) has been preferred to general anaesthesia in specialist centres, this practice has not been reflected in general hospitals, where most operations occur. This meta-analysis will analyse differences in outcome between the two techniques from previous studies conducted in general hospitals to determine the evidence for practice in this setting. MATERIALS AND METHODS: A search of PubMed, Medline and Embase was undertaken, as well as a search for UK presentations, limited to English language trials published since 1966. This identified five randomised controlled trials and a total of 895 patients for meta-analysis. Only outcomes reported in at least three studies were included. RESULTS: The number of patients experiencing postoperative nausea was significantly reduced in the LA group. The time taken to return to normal social activities was also significantly reduced in the LA group, by a period of 1 day. There were no differences in urinary retention rates or return to work. The time in theatre and the operative time were not altered significantly by differing anaesthetic techniques. CONCLUSIONS: Meta-analysis of the five trials comparing local and general anaesthesia for inguinal hernia repair fails to confirm the wide-ranging benefits reported by the individual studies. Local anaesthetic reduces nausea and accelerates return to normal activities following open inguinal hernia repair. The benefit of LA is sufficiently small that its use should be dictated by patient and clinician preference.

Do teenagers return to normal physical activity levels after limb fractures? A longitudinal, accelerometry-based, activity monitoring study.

PURPOSE: This study aimed to elucidate whether levels of physical activity (PA) return to normal after bone healing or whether long-term behavioural changes in PA are to be expected in children and teenagers who have sustained limb fractures. METHODS: In all, 100 children and teenagers with a first episode of limb fracture and 100 sex- and age-matched healthy controls (CTRL) were recruited for a prospective study. PA in limb fracture patients was assessed at 18-month follow-up using accelerometer measurements, and values were compared with those of CTRL. Time spent in PA at different levels of intensity was determined for each participant and expressed in minutes and as a percentage of total validly measured time. RESULTS: Mean levels of PA at different levels of intensity by previously injured children and teenagers were similar than CTRL (42 sets of paired data). However, time spent in moderate-to-vigorous PA (MVPA) was lower than 60 minutes among limb-fracture patients at 18-month follow-up. CONCLUSION: The amount of skeletal loading in children and teenagers returns to normal values by 18 months after limb fracture. Even if time spent in MVPA is not significantly lower in children and teenagers with limb fractures, it no longer reached the international recommendations for school-aged children (MVPA > 60 minutes), which may be interpreted as a lifestyle modification or a behavioural change to avoid new trauma. LEVEL OF EVIDENCE: II.

An LC method for the simultaneous screening of some common counterfeit and sub-standard antibiotics Validation and uncertainty estimation.

Pharmaceutical counterfeiting is a worldwide public health problem, often under-recognised, especially in developing countries where the percentage of counterfeit and sub-standard medicines is dramatically high. Antibiotics, among the most widespread drugs, have been particularly targeted by counterfeiters. World Health Organization emphasizes the need for development and distribution of screening methods explicitly targeted to counterfeit drugs. In this paper is presented a single method for the simultaneous analysis of some of the most common and counterfeited essential antibiotics: ampicillin, amoxicillin+clavulanic acid, doxycycline, cloxacillin, chloramphenicol. A full validation was performed in terms of linearity, precision, robustness and trueness; an assessment of uncertainty was carried out exploiting these data. A wide linearity range was investigated considering the specific nature of counterfeit and sub-standard drugs, whose content in active substance may be rather far from the declared amount. A large span in robustness parameters was considered and a complete intermediate precision assessment was conducted, envisaging the possibility of transferring the method to quality control laboratories, hopefully in developing countries. Finally, the method was successfully applied to the analysis of antibiotics purchased on the informal market in Chad, among which counterfeit and sub-standard samples were detected.

Response to Alpha-Interferon Treatment of the Congenital Dyserythropoietic Anemia type I in Two Sicilian Beta Thalassemia Carriers.

Congenital dyserythropoietic anemia type I (CDAI) is an autosomal recessive inherited haematological disorder associated with moderate-to-severe anemia characterized by ineffective erythropoiesis with distinct morphological abnormalities in erythroid precursors. We present two case of congenital dyserythropoietic anemia type I in two Sicilian patients heterozygous for ß0 39 globin gene cod 39 C > T with marked bone marrow abnormalities, responding to treatment with alpha interferon. The diagnosis was established using routine haematological and biochemical test, light and electron microscopy; molecular analysis of the CDAN1 gene associated to the CDAI disease was performed. The response to the treatment was monitored using the hemoglobin levels, the red cell count, the reticulocyte count and the transfusional requirement. This report points out the usefulness of the treatment with interferon alpha in two Sicilian beta thalassemia carriers, in which the therapy was well tolerated without producing any side effects; in these patients the transfusion requirements after the initiation of interferon therapy decreased.

[Intra-articular treatment with the TNF-alpha antagonist, etanercept, in severe diffuse pigmented villonodular synovitis of the knee]. / Trattamento intra-articolare con l'antagonista del TNFalpha etanercept nella sinovite villo-nodulare pigmentosa diffusa del ginocchio.

Pigmented villonodular synovitis (PVNS) is a rare pre-malignant disease that require aggressive treatment as surgical synovectomy, eventually followed by radiosynovectomy. Nevertheless, the disease often reoccurs after these treatments. To determine the safety and efficacy of intra-articular (IA) TNFalpha blockade with etanercept (ETN), before extended arthroscopic synovectomy, in severe PVNS of the knee, two patients, (a 26-year-old man with B27+ undifferentiated spondylarthropathy and a 32-year-old femal with seronegative oligoarthritis), affected by diffuse knee PVNS (diagnosis made by histological examination), resistant to IA corticosteroid injections and to repeated arthroscopic synovectomy, were submitted, after protocol approval by human research committee and patient's written informed consent to intra-articular etanercept (IA-ETN) treatment with a different dosage schedule: 12.5 mg weekly IA-ETN injection for 4 weeks, followed by extended arthroscopic synovectomy and of 25 mg IA-ETN injection for 4 weeks, respectively. Previous DMARDs treatment was continued in stable appropriate doses. Any adverse events were recorded throughout the study. The following parameters were considered as clinical endpoints: 1) Knee Joint Index (KJI: range 0-14); 2) Thompson index (THI: range 0-9) At the study entry and at the end of follow-up, high frequency ultrasound grey scale synovial thickening (US-ST) was also assessed. No adverse events were observed due to IA-ETN and to arthroscopic synovectomy. Marked improvement of knee disease activity over time and sustained functional recover was obtained. US-ST evaluation before treatment initiation and at the end of follow-up confirmed the regression of knee joint synovial proliferation.

New Codanin-1 Gene Mutations in a Italian Patient with Congenital Dyserythropoietic Anemia Type I and Heterozygous Beta-Thalassemia.

Congenital dyserythropoietic anemia type I is an autosomal recessive disorder associated with macrocytic anemia, ineffective erythropoiesis, iron overloading and characterized by abnormal chromatin ultrastructure in erythroblasts such as internuclear chromatin bridges, spongy heterochromatin and invagination of the nuclear membrane. A 58-year-old Causasian man with chronic hemolytic anemia, heterozygous for ß (+) -globin IVS1, nt110 G>A mutation (causing abnormal alpha:beta globin chain ratio) showed clinical, laboratory and hematological features suggesting diagnosis of CDA1. Sequence analysis of CDA-related genes revealed compound heterozygosity for two novel mutations in the CDAN1 gene: a frameshift mutation 3367 del 4 (TTAG) in exon 25 and a missense mutation c.1811 G>T in exon 11 causing an aminoacid change from glycine to valine at codon 565 (G565V). One of the propositus' brothers showed the same gene mutations. As the CDA1 can mimic thalassemia, a frequent misdiagnosis is possible especially in countries where the prevalence of thalassemia is high. A strong clinical suspicion in patients who do not reveal a clear genetic basis for presumed thalassemia may help clinch the correct diagnosis.

Quality indicators in the intensity modulated/image-guided radiotherapy era.

PURPOSE: To propose new Quality Indicators (QIs) for the Intensity Modulated(IMRT)/Image-Guided(IGRT) Radiotherapy techniques. MATERIALS AND METHODS: Two structure, 10 process and 2 outcome QIs were elaborated. A working group including Radiation Oncologist, Medical Physicist and Radiation Technologists was made up. A preliminary set of indicators was selected on the basis of evidenced critical issues; the criteria to identify more relevant and specific QIs for IMRT/IGRT were defined; structure, process and outcome QIs were defined. The elaborated indicators were tested in four Italian Radiotherapy Centers. RESULTS: Fourteen indicators were proposed. Seven indicators were completely new while a new standard is proposed for four indicators based on Validation Centers (VC) data. No change was reported for 3 indicators. The indicators were applied in the four VC. The VC considered were able to respect all indicators except indicator 2 for one Center. DISCUSSION AND CONCLUSION: QIs may provide useful measures of workload and service performances.

Co-heredity of silent CAP + 1570 T>C (HBB:c*96T>C) defect and severe ß-thal mutation: a cause of mild ß-thalassemia intermedia.

INTRODUCTION: During an intensive screening program aimed at identifying the healthy carriers of thalassemia and the couples at risk of bearing an affected fetus, a rare single nucleotide variation (SNV), CAP + 1570 T > C (HBB:c*96T > C), located 12 nucleotides upstream of the polyadenylation signal in 3'UTR of the beta globin gene was identified. It was previously reported as a ß+ thalassemia mutation and later as a plain polymorphism. METHODS: Genotype identification of globin gene mutations was carried out using sequencing analysis, GAP-PCR, and MLPA methods. RESULTS: CAP + 1570 T > C (HBB:c*96T > C) was found in 39 heterozygotes, in one case in homozygous state and in thirteen cases of co-inheritance of this nucleotide substitution with other mutations in globin genes. Carriers of this mutation showed a 'silent' phenotype without appreciable microcytosis and hypochromia, so they cannot be differentiated from noncarrier individuals. Compound heterozygotes for this mutation and severe ß-thal mutations showed a variable phenotype ranging from ß-thal carrier to mild form of ß-thalassemia intermedia, revealing new aspects and allowing to better understand the clinical implications of this nucleotide substitution that can be classified as a silent ß-thalassemic defect. CONCLUSION: Data reported in this study indicate the need of investigating partner of ß-thalassemia carrier by complete sequencing analysis of ß-globin gene and of providing an appropriate genetic counseling for couples at risk undergoing prenatal diagnosis.

Effects of Mercuric Chloride on the Hydraulic Conductivity of Tomato Root Systems (Evidence for a Channel-Mediated Water Pathway).

A pressure-flux approach was used to evaluate the effects of HgCl2 on water transport in tomato (Lycopersicon esculentum) roots. Addition of HgCl2 to a root-bathing solution caused a large and rapid reduction in pressure-induced root water flux; the inhibition was largely reversible upon addition of [beta]-mercaptoethanol. Root system hydraulic conductivity was reduced by 57%. There was no difference between treatments in the K+ concentration in xylem exudate. The results are consistent with the presence of a protein-mediated path for transmembrane water flow in tomato roots.

Infection with Anaplasma phagocytophilum in a seronegative patient in Sicily, Italy: case report.

BACKGROUND: Anaplasma phagocytophilum causes human granulocytic anaplasmosis (HGA) in humans, which has been recognized as an emerging tick-borne disease in the United States and Europe. Although about 65 cases of HGA have been reported in Europe, some of them do not fulfill the criteria for confirmed HGA. Confirmation of HGA requires A. phagocytophilum isolation from blood, and/or identification of morulae in granulocytes and/or positive PCR results with subsequent sequencing of the amplicons to demonstrate specific rickettsial DNA. Seroconversion or at least fourfold increase in antibody titers to A. phagocytophilum has been used as criteria for confirmed HGA also. CASE PRESENTATION: Infection with A. phagocytophilum was confirmed by PCR in a patient in Sicily, Italy, who had negative serology for A. phagocytophilum. A fragment of A. phagocytophilum 16S rDNA was amplified by two independent laboratories and sequenced from two separate patient's blood samples. The 16S rDNA sequence was identical in both samples and identical to the sequence of the A. phagocytophilum strain USG3 originally obtained from a dog. CONCLUSION: Infection with A. phagocytophilum was confirmed in a patient without a detectable antibody response against the pathogen. The results reported herein documented the first case of confirmed HGA in Sicily, Italy. These results suggested the possibility of human infections with A. phagocytophilum strains that result in clinical symptoms and laboratory findings confirmatory of HGA but without detectable antibodies against the pathogen.