Multi-approach analysis of the diversity in Colletotrichum cliviae sensu lato.

Colletotrichum cliviae is a fungal species reported both as pathogen and endophyte with broad geographical distribution. Some purported isolates of this species have been assigned to different taxa, including Colletotrichum aracearum, Colletotrichum orchidearum and Colletotrichum. sichuanensis, for which a preliminary analysis of extensive multilocus (ACT, GAPDH, ITS, TUB2) data in this study revealed high sequence similarity with C. cliviae. We further reassessed the species delineation by using the coalescent method of the generalized mixed Yule-coalescent (GMYC) and Poisson Tree Processes (PTP). Single and multilocus gene trees strongly supported a C. cliviae s. lat. clade including the four species. This clade unfolded eight subclades grouped into three distinct lineages, but no monophyly of any of the four species. GMYC and PTP analyses confidently supported the evolutionary independence of these lineages. C. sichuanensis and C. cliviae, except one isolate, formed the largest lineage. The second lineage was made up of isolates named C. aracearum and some of C. orchidearum sharing the haplotype and the third lineage accommodated two isolates named C. cliviae and C. orchidearum. This finding suggests the synonymization of C. sichuanensis with C. cliviae whereas the taxonomic status of C. aracearum and C. orchidearum still needs clarification. This study lays great stress upon the use of comprehensive data for sequence-based characterisation of species in the C. cliviae s. lat. It also presents the first report of C. cliviae in tropical Africa and on citrus host.

A facile method for urinary phenylalanine measurement on paper-based lab-on-chip for PKU therapy monitoring.

A miniaturized paper-based lab-on-chip (LoC) was developed for the facile measurement of urinary Phe (phenylalanine) level on PKU (Phenylketonuria) treated patient. This system permits the monitoring of Phe in a dynamic range concentration of 20-3000 µM.

Long-term use of carglumic acid in methylmalonic aciduria, propionic aciduria and isovaleric aciduria in Italy: a qualitative survey.

OBJECTIVE: Organic acidurias (OAs) are a group of rare metabolic disorders that disrupt the regular amino acid metabolism. OAs are characterized by recurrent episodes of acidemia, ketonuria and hyperammonemia which can result in brain/liver damage and renal failure, and despite the life-long protein-restricted diet, impaired growth and long-term complications can occur. Consequently, a long-term management of OAs patients is required, aimed principally at reducing the frequency and duration of metabolic decompensation/hyperammonemia episodes. Nevertheless, unlike the acute phase, evidence on the chronic management of OAs patients is less consolidated. SUBJECTS AND METHODS: To expand the knowledge on this field, 13 Italian referral centers for the management of OAs were involved in a survey focused on the long-term use of carglumic acid (Carbaglu®, Recordati Rare Diseases). RESULTS: Participating centers reported a reduction between 69% and 81% in the annual number of metabolic decompensations with the chronic use of carglumic acid and an improvement in protein intake. Most centers reported no difficulty using carglumic acid as a long-term therapy, along with a great compliance. CONCLUSIONS: Taken together, obtained data align with the available literature and support a positive clinical experience with the long-term carglumic acid administration. Additional studies aimed at better defining a proper dosage for the chronic administration of carglumic acid and the clinical and biochemical characteristics of patients treated chronically are needed. In addition, the potential impact of this treatment regimen on the neurological development and growth of patients should be elucidated.

Phenotypic variability, neurological outcome and genetics background of 6-pyruvoyl-tetrahydropterin synthase deficiency.

This study aimed to investigate the clinical variability and factors implied in the outcome of 6-pyruvoyl-tetrahydropterin synthase deficiency (PTPSd). Biochemical and clinical phenotype, treatment variables, and 6-pyruvoyl-tetrahydropterin synthase (PTS) genotype, were explored retrospectively in 19 Italian patients (12 males and 7 females, aged 4 months to 33 years). According to the level of biogenic amines in cerebrospinal fluid (CSF) at the diagnosis, the patients were classified as mild (6) (normal level) or severe (13) (abnormal low level) form (MF and SF, respectively). Blood Phe ranged from 151 to 1053 micromol/l in MF (mean +/- SD: 698 +/- 403) and 342-2120 micromol/l in SF (mean +/- SD: 1175 +/- 517) (p = 0.063). Patients with MF showed a normal neurological development (a transient dystonia was detected in one), while all SF patients except one presented with severe neurological impairment and only four had a normal neurological development. The outcome of the SF was influenced by the precocity of the treatment. Serial CSF examinations revealed a decline of 5-hydroxyindolacetic acid in MFs and an incomplete restoration of neurotransmitters in SFs: neither obviously affected the prognosis. PTS gene analysis detected 17 different mutations (seven so far unreported) (only one affected allele was identified in three subjects). A good correlation was found between genotype and clinical and biochemical phenotype. The occurrence of brain neurotransmitter deficiency and its early correction (by the therapy) are the main prognostic factors in PTPSd.

Chromium (VI) reduction in activated sludge bacteria exposed to high chromium loading.

A mixed-culture of bacteria collected from a wastewater treatment plant in Brits, North-West Province (South Africa) biocatalytically reduced Cr(VI) at much higher concentrations than previously observed in cultures isolated in North America. Complete Cr(VI) reduction in aerobic cultures was achieved at a high concentration of 200 mg/L after incubation for only 65 hours. Under anaerobic conditions up to 150 mg, Cr(VI)/L was completely removed after incubating for 130 to 155 hours, still higher than the Cr(VI) reduction achieved with previous cultures where complete removal was only observed in cultures with the added Cr(VI) concentration not greater than 30 mg/L. Cr(VI) reduction capability of the cultures was verified in purified cultures. Consortium cultures were characterised using 16S rRNA partial sequence analysis. Results showed that the gram-positive Bacillus genera predominated under aerobic conditions with a small composition of the gram-negative Microbacterium sp. There was more biodiversity observed in the anaerobic cultures with the marked appearance of Enterococcus, Arthrobacter, Paenibacillus and Oceanobacillus species. The results showed that Cr(VI) reduction rate in the new culture was up to eight times higher than that previously observed in other Cr(VI) reducing cultures isolated from Cr(VI) contaminated soil environments in Newark (New Jersey) and other sites in North America.

Levels of L-asparagine in CSF after intramuscular administration of asparaginase from Erwinia in children with acute lymphoblastic leukemia.

PURPOSE: As part of a study on the pharmacokinetics associated with the administration of asparaginase (ASNase) from Erwinia to the CNS, we determined the levels of asparagine in the CSF of children with acute lymphoblastic leukemia (ALL). PATIENTS AND METHODS: Twenty children received eight standard doses of intramuscular ASNase (10,000 IU/m2) every 3 days as part of induction therapy. In the postremission phase of therapy, the children were randomized to receive either 20 courses of high-dose intramuscular ASNase (25,000 IU/m2) weekly (n = 8) or four courses of standard-dose intramuscular ASNase (10,000 IU/m2) every 3 days (n = 12). RESULTS: All patients had detectable levels of L-asparagine in the CSF at the time of diagnosis. The levels of L-asparagine in CSF were undetectable in 15 of 20 (75%) and in seven of 19 (36.8%) children 3 and 5 days, respectively, after administration of standard-dose ASNase. After administration of high-dose ASNase, the levels of L-asparagine in the CSF were undetectable in five (62.5%) and two (25%) of eight children after 3 and 5 days, respectively. CONCLUSION: In this study 60% to 70% and 25% to 35% of children had complete depletion of L-asparagine from the CSF after 3 and 5 days, respectively, after administration of both schedules of ASNase from Erwinia. In the remaining patients, administration of ASNase may have resulted in a suboptimal antileukemic effect at the CNS level.

Clinical and molecular findings in hyperornithinemia-hyperammonemia-homocitrullinuria syndrome.

The authors report the clinical and molecular findings in eight patients with hyperornithinemia, hyperammonemia, and homocitrullinuria (HHH) syndrome. The most consistent neurologic finding was spastic paraparesis, seen in five of the eight patients. However, all showed signs of pyramidal tract involvement. A broad spectrum of pathogenetic mutations (including missense, nonsense, splice site, insertion, and deletions) were identified in the ORNT1 gene.

Neurotransmitter amino acid levels in rat thalamus and cerebral cortex after cerebellectomy.

Glutamate, aspartate, GABA, glycine and taurine levels have been measured in rat thalamus and in cerebral cortex at different time intervals (3rd, 7th, 15th, 30th day) after cerebellectomy. A decrease in glutamate, aspartate and GABA was detected at the 7th day after cerebellectomy in the thalamus and at the 15th day in the cerebral cortex; at the 30th day after cerebellectomy the levels of these amino acids in the thalamus and in the cerebral cortex were observed to have recovered to control values. No statistically significant difference in glycine and taurine levels in the thalamus and in the cerebral cortex after cerebellectomy could be seen. These results show that the functional recovery process after cerebellar injury is associated with a complex modification of amino acid levels in thalamus and in cerebral cortex.

Activity of 1,2-dimethylhydrazine in the mouse bone marrow micronucleus assay using a triple- and a single-dosing protocol.

In the present study the ability of 1,2-dimethylhydrazine (DMH) to induce micronuclei in mouse bone marrow erythrocytes was investigated using 2 different dosing regimens. DMH caused an increase in micronuclei in both male and female mice following single administration and sampling after 24 h. The effect was more pronounced in female than in male animals. Triple administration of DMH at concentrations corresponding to 80, 40 and 20% of the median lethal dose (MLD) did not increase the incidence of micronuclei in either sex. Small increases in micronucleus incidence were observed after triple dosing at 100% of the MLD value. These results suggest that a future micronucleus protocol should include animals of both sexes and single and repeated administration of the test substance.

Cytogenetic effects of benzimidazoles in mouse bone marrow.

The cytogenetic effects of three benzimidazoles, i.e., benomyl, methyl thiophanate and methyl 2-benzimidazolecarbamate (MBC), were studied in mouse bone marrow cells by analyzing three genetic endpoints: micronuclei, structural chromosome aberrations plus or minus gaps, and aneugenic effects (hyperdiploidy or polyploidy). In general, the effects were small, but it was observed that benomyl and MBC significantly induced micronuclei as well as aneugenic effects, hyperdiploidy (no metaphases with more than one or two extra chromosomes, 2n + 1 or 2n + 2, were observed) and polyploidy (4n). The induction of chromosome gaps and breaks was less evident. Methyl thiophanate significantly induced micronuclei, but it was less effective than benomyl and MBC. Our results showed that micronuclei are a good indicator of aneugenic effects in mouse bone marrow cells. A curvilinear trend test has been devised to fit the curves originating from the time-dependent responses.

[Surgical treatment of non-toxic multinodular goitre. Retrospective evaluation of a personal case series]. / La terapia chirurgica del gozzo multinodulare non tossico. Valutazione retrospettiva di una casistica personale.

The authors base their study on a retrospective evaluation of a personal series of patients and on a review of the literature on this topic. They focus their attention on surgical problems relating to the type of treatment of non-toxic multinodular goitre. On the basis of the data obtained, they underline that in this pathology total thyroidectomy is the most appropriate form of surgery and does not entail a higher morbidity in relation to more conservative resection.

[The echographic aspects of the splenoportal axis in patients with esophageal varices]. / Aspetti ecografici dell'asse spleno-portale in pazienti con varici esofagee.

In the present study the A. try to assess the usefulness of ultrasound in detecting blending risk in portal hypertension. Endoscopic and sonographic findings of 35 cirrhotic patients with esophageal varices were compared to a control group of 27 hepatopathy patients without portal hypertension, observed in the same department and in the same interval. The medium caliber of portal vein was 14.64 +/- 2.59 and 11.8 +/- 1.33 mm, respectively in the study and in the control group (p less than 0.01). Statistic correlations were also found in subgroups of patients with varices of different degrees. A portal vein greater than 16 mm was found in 10 of 11 patients with large varices; this pattern provides a sensitivity of 71.4% and a specificity of 94.1%. In our experience this echographic finding should represents a clear indication for close endoscopic follow-up, even in patients without previous bleeding from esophageal varices.

[Nonpalpable lesions of the breast. A new technique of ultrasound-guided excision biopsy]. / Le lesioni non palpabili della mammella. Una nuova tecnica di biopsia escissionale ecoguidata.

The authors have developed a simple and accurate technique of excisional biopsy for non-palpable breast nodes using an intraoperative sonographic technique. Identification, localization and exeresis of the lesions are guided by means of a sterilized transducer introduced through the surgical wound. The confirmation of the correct biopsy is evaluated by scanning the specimen gowned with a surgical glove and comparing the intra- and postoperative ultrasonographic images. The results of analysis showed that in all cases the altered area was removed with extreme accuracy and the ultrasonography is reliable in identifying pathological or suspicious areas since the dimensions of the lesion measured with the scanner correspond significantly to the dimensions of the lesions when measured histologically.

[Acute abdominal pain in emergency surgery. Clinical epidemiologic study of 450 patients]. / Il dolore addominale acuto in chirurgia d'urgenza. Studio clinico-epidemiologico su 450 pazienti.

A retrospective study on 450 patients admitted in emergency with acute abdominal pain in a general surgical unit of the Ist Institute of Surgery of the University of Rome "La Sapienza" was carried out during a 7 years period. The aim of the study was to identify the most frequent causes of acute abdominal pain correlated with age, sex, onset of pain and other conditions in order to improve diagnosis and reduce mobility for mortality. Appendicitis was the most frequent diagnosis (75 pts = 16.4%). Non-specific abdominal pain (NSAP), found in 71 pts (15.5%), cholelithiasis in 57 pts (12.5%), abdominal malignancy in 47 pts (10.3%) were other frequent conditions. The largest number of admissions occurred in the age groups 60-70 years (16.6%) and 20-30 years (14.2%). Surgical operations were performed in 206 patients (45.7%). The overall mortality was 4.2% (19/450 pts) and the rate increased significantly in patients aged > 60 years. Postoperative mortality was 5.8% (12/206 pts) while mortality rate in non-operated patients was 2.8%. The causes of perioperative death included perforated peptic ulcer, abdominal malignancies (15.4%) and urgent colonic resections (9.4%). The duration of inpatients stay increased significantly with the age of the patients, including those with nsap. The results of the study indicate a need to review the methods of diagnosis of appendicitis and to obtain a better clinical performance in patients with nsap.

[Role of ultrasonography in emergency surgery]. / Ruolo dell'ecografia in chirurgia d'urgenza.

A prospective study was carried out to compare the value of ultrasound examination with conventional management in patients presenting with emergency surgical problems. During an 4 years period the authors followed 301 patients referred to the Ist Institute of Surgery of the University of Rome "La Sapienza". After immediate clinical evaluation one or more US examinations were performed in each patient following a standard protocol of study. Abdominal emergencies were the most frequent conditions, found in 75% of the cases and the acute abdomen due to trauma in 17.6% of the cases. The overall sensibility, specificity and accuracy of clinical examinations and US examinations were respectively of 88%, 33%, 70% and 95%, 94%, 95%. In 38 patients (12.7%), a diagnosis was made up by sonographic examination which had not been clinically expected (group A). The principal condition in this group was gynaecological pathology, found in 17 patients. In 161 patients (53.3%) sonography confirmed the first diagnosis clinically suspected (group B) and in 23 cases the second or third differential diagnosis (group D). Sonography made no contribution to the diagnosis in 77 cases (25,5%) and only in 2 cases was considered misleading. The result of this study demonstrate the usefulness of emergency ultrasonography in gynecological, gall bladder, pancreatic and vascular diseases. Moreover sonography must be considered the principal diagnostic tool in patients with trauma. In order to recognize gynecological diseases which can simulate appendicitis US examination must be considered necessary in young female patients. When appendicitis is suspected US is useful in children and in older patients too.

[Genetic role in hernia of the esophageal hiatus. Description of the disease in 2 brothers]. / Ruolo genetico nelle ernie dello jatus esofageo. Descrizione dell'affezione in due fratelli.

Two brothers born 15 years apart from normal healthy parents were affected by esophageal hiatus hernia diagnosed at 4 and 8 months respectively. We found 37 other families with more than one member affected by EHH reported in the literature. Since the anomaly often runs asymptomatic, familiarity is probably more frequent than generally thought. The type of inheritance is still uncertain, but a multifactorial etiology is the most likely explanation.

[Megaloblastic anemia: a unusual complication in an infant with double allergy to both cow's milk protein and soy]. / Anemia megaloblastica: singolare complicanza in lattante con doppia allergia alle proteine del latte vaccino ed alla soja.

The authors reported a case of a child suffering from Cow's milk protein allergy, in which a soybean milk was administered for one month. The incorrect preparation of the formula determined a megaloblastic anaemia due to folate deficiency. The observation of very high levels of seric aminoacids during the acute phase of the disease was noteworthy. The therapy with folic acid and the introduction of protein hydrolysated formula milk determined a rapid normalization of the haematological picture. During the follow-up the administration of soybean protein caused anaphylaxis showing the contemporary presence of soybean allergy. The challenge test with cow's milk performed afterwards showed the restoration of the tolerance towards cow's milk protein.

[Neuronal ceroid lipofuscinosis. Description of a case]. / Le ceroido-lipofuscinosi neuronali. Descrizione di un caso.

The Authors describe a case of Batten-Bielschowsky disease and how they made the diagnosis. It's stressed the importance of the pathological examination, in the absence of any laboratory tests for the diagnosis of this disease.

[Hemorrhagic complication following an external shunt for hemodialysis: a clinical case and a proposed technical procedure for its prevention]. / Complicanza emorragica post shunt esterno per emodialisi: caso clinico e proposta di un accorgimento tecnico di prevenzione.

The vascular access for haemodialysis is still a problem not completely solved although surgical experience started more than thirty years ago. Particularly haemorrhagic complications can be dangerous because many patients go home after haemodialysis. The authors report a case in which the disconnection of the conic tip from the shunt cannula for haemodialysis induced an external bleeding, and propose a safety ligature. However, the latter is not necessary for a cannula integrated with the vessel tip.

[A case of carcinoid with multiple sites in the jejunum and distal ileum]. / Un caso di carcinoide a localizzazioni multiple su digiuno ed ileo distale.

Multiple carcinoid tumors of the small bowel with more than 3 lesions are very unusual. The authors report a case with 4 lesions, 2 of those localized in the jejunum with more advanced infiltration of the wall and extension to regional mesenteric lymph nodes, revealed by ultrasonography. The relative low incidence and particularly the vague, nonspecific clinical presentation, the unusual site in the jejunum, and failure of the radiological examine of one year before lead to not suspect this condition prior to US examination. However, the feature of asymmetric, concentric thickening of the bowel wall requiring a more accurate exam by CT with oral contrast was able to confirm the suspect of the intestinal tumor. The patient, 80 year old, underwent radical surgery with a wide lymph nodes dissection as well as double resection of the jejunum and distal ileum. The post-surgical outcome was uneventful. A 12-month follow-up is free of the disease.