Production of laccase by Botrytis cinerea and fermentation studies with strain F226

After induction, seven strains ofBotrytis cinerea released into the culture broth considerable amounts of laccase in a brief production time. The set-up of a suitable production process was studied with a selected strain in a 10-L fermenter. The optimum fermentation conditions were a 3% inoculum with a high degree of sporulation, a simple medium containing 20 g L−1 of glucose and 2 g L−1 of yeast extract at pH 3.5, 2 g L−1 gallic acid as inducer, added after 2 days of growth, an agitation speed of 300 rpm, an aeration rate of 1.2 vvm and a temperature of 24°C. By optimizing the culture conditions, the enzyme activity reached 28 U ml−1 in 5 days with a specific activity of 560 U mg−1 protein. The best procedure to obtain a suitable crude enzyme preparation was concentration of the supernatant medium to 10% of the initial volume by ultrafiltration, followed by a fractional precipitation with ethanol. The optimum pH and temperature for laccase activity were 5.5 and 40°C, respectively, with syringaldazine as the substrate

Characterization of multiple enhancer regions upstream of the apolipoprotein(a) gene.

Plasma concentrations of the atherogenic lipoprotein(a) (Lp(a)) are predominantly determined by inherited sequences within or closely linked to the apolipoprotein(a) gene locus. Much of the interindividual variability in Lp(a) levels is likely to originate at the level of apo(a) gene transcription. However, the liver-specific apo(a) basal promoter is extremely weak and does not exhibit common functional variations that affect plasma Lp(a) concentrations. In a search for additional apo(a) gene control elements, we have identified two fragments with enhancer activity within the 40-kilobase pair apo(a)-plasminogen intergenic region that coincide with DNase I-hypersensitive sites (DHII and DHIII) observed in liver chromatin of mice expressing a human apo(a) transgene. Neither enhancer exhibits tissue specificity. DHIII activity was mapped to a 600-base pair fragment containing nine DNase I-protected elements (footprints) that stimulates luciferase expression from the apo(a) promoter 10–15-fold in HepG2 cells. Binding of the ubiquitous transcription factor Sp1 plays a major role in the function of this enhancer, but no single site was indispensable for activity. DHIII comprises part of the regulatory region of an inactive long interspersed nucleotide element 1 retrotransposon, raising the possibility that retrotransposon insertion can influence the regulation of adjacent genes. DHII enhancer activity was localized to a 180-base pair fragment that stimulates transcription from the apo(a) promoter 4–8-fold in HepG2 cells. Mutations within an Sp1 site or either of two elements composed of direct repeats of the nuclear hormone receptor half-site AGGTCA in this sequence completely abolished enhancer function. Both nuclear hormone receptor elements were shown to bind peroxisome proliferator-activated receptors and other members of the nuclear receptor family, suggesting that this enhancer may mediate drug and hormone responsiveness

Robotic prostatectomy : an update on functional and oncologic outcomes

Since the first procedure performed in 2000, robotic-assisted radical prostatectomy (RARP) has been rapidly gaining increasing acceptance from both urologists and patients. Today, RARP is the dominant treatment option for localised prostate cancer (PCa) in the United States, despite the absence of any prospective randomised trial comparing RARP with other procedures. Robotic systems have been introduced in an attempt to reduce the difficulty involved in performing complex laparoscopic procedures and the related steep learning curve. The recognised advantages of this kind of minimally invasive surgery are three-dimensional (3D) vision, ten-fold magnification, Endowrist technology with seven degrees of freedom, and tremor filtration. In this article, we examine this technique and report its functional (in terms of urinary continence and potency) and oncologic results. We also evaluate the potential advantages of RARP in comparison with open and laparoscopic procedures. Copyright

tolterodine improves quality of life in patients with an overactive bladder with or without urinary incontinence a prospective multicenter study

AIMS: In order to analyse the effect of tolterodine on the Quality of life (QoL) of patients with overactive bladder (OB) we conducted a prospective multicentre clinical study. MATERIALS AND METHODS: Subjects were questioned at entry and 4, 12 and 24 weeks later about the number of micturitions and incontinent and urgency episodes/day, using a micturition diary. The mean volume voided per micturition and the number of pads used per day was also recorded. The QoL was measured using the Kings Health Questionnaire (KHQ) and the Incontinence Impact Questionnaire (IIQ). A total of 179 patients entered the study: 59 dropped out (4 due to lack of efficacy, 10 due to adverse events, 25 because of lack of interest in the study/other reason and 20 were lost at follow up), leaving 120 patients for analysis. One hundred and eight patients (90%) were female, their mean age was 56.5 years (SD 11.2); 87 had never received treatment for OB/UI (80.6%) and their mean weight was 70.0 Kg (SD 12.7). RESULTS: The mean number of micturitions/day was 9.3 at trial entry and it decreased to 6.8 by the end of the study. The corresponding values for the number of urge episodes, incontinence episodes and number of pads used per day were 3.5, 2.7 and 1.2 and 0.8, 0.9 and 0.4 respectively. The mean volume voided per micturition increased from 146 ml. to 178 ml. All the differences between trial entry and end of study values were statistically significant (p<0.05). Considering the results of the KHQ, the values of all the different areas/domini (?) decreased markedly and in a statistically significant way between the start of treatment and the end of study evaluations. Similar findings emerged when we considered values of the IIQ. The decrease was constant and marked during the first three months and remained constant thereafter. CONCLUSIONS: This study, conducted in a population of subjects with dry and wet OB, shows that tolterodine given for six months lowers the frequency of urgency episodes and incontinence episodes without troublesome adverse effects. These clinical effects are mirrored in the QoL, KHQ and IIQ questionnaire scores, which improved by about 50% over the same period

Donor cell acute myeloid leukemia after hematopoietic stem cell transplantation for chronic granulomatous disease: a case report and literature review

The patient reported here underwent hematopoietic stem cell transplantation (HSCT) due to chronic granulomatous disease (CGD) caused by biallelic mutations of the NCF1 gene. Two years later, he developed AML, which was unexpected and was recognized via sex-mismatched chromosomes as deriving from the donor cells; the patient was male, and the donor was his sister. Donor cell leukemia (DCL) is very rare, and it had never been reported in patients with CGD after HSCT. In the subsequent ten years, the AML relapsed three times and the patient underwent chemotherapy and three further HSCTs; donors were the same sister from the first HSCT, an unrelated donor, and his mother. The patient died during the third relapse. The DCL was characterized since onset by an acquired translocation between chromosomes 9 and 11, with a molecular rearrangement between the MLL and MLLT3 genes-a quite frequent cause of AML. In all of the relapses, the malignant clone had XX sex chromosomes and this rearrangement, thus indicating that it was always the original clone derived from the transplanted sister's cells. It exhibited the ability to remain quiescent in the BM during repeated chemotherapy courses, remission periods and HSCT. The leukemic clone then acquired different additional anomalies during the ten years of follow-up, with cytogenetic results characterized both by anomalies frequent in AML and by different, non-recurrent changes. This type of cytogenetic course is uncommon in AML

Association of kidney disease measures with risk of renal function worsening in patients with type 1 diabetes

Background: Albuminuria has been classically considered a marker of kidney damage progression in diabetic patients and it is routinely assessed to monitor kidney function. However, the role of a mild GFR reduction on the development of stage 653 CKD has been less explored in type 1 diabetes mellitus (T1DM) patients. Aim of the present study was to evaluate the prognostic role of kidney disease measures, namely albuminuria and reduced GFR, on the development of stage 653 CKD in a large cohort of patients affected by T1DM. Methods: A total of 4284 patients affected by T1DM followed-up at 76 diabetes centers participating to the Italian Association of Clinical Diabetologists (Associazione Medici Diabetologi, AMD) initiative constitutes the study population. Urinary albumin excretion (ACR) and estimated GFR (eGFR) were retrieved and analyzed. The incidence of stage 653 CKD (eGFR < 60 mL/min/1.73 m2) or eGFR reduction > 30% from baseline was evaluated. Results: The mean estimated GFR was 98 \ub1 17 mL/min/1.73m2 and the proportion of patients with albuminuria was 15.3% (n = 654) at baseline. About 8% (n = 337) of patients developed one of the two renal endpoints during the 4-year follow-up period. Age, albuminuria (micro or macro) and baseline eGFR < 90 ml/min/m2 were independent risk factors for stage 653 CKD and renal function worsening. When compared to patients with eGFR > 90 ml/min/1.73m2 and normoalbuminuria, those with albuminuria at baseline had a 1.69 greater risk of reaching stage 3 CKD, while patients with mild eGFR reduction (i.e. eGFR between 90 and 60 mL/min/1.73 m2) show a 3.81 greater risk that rose to 8.24 for those patients with albuminuria and mild eGFR reduction at baseline. Conclusions: Albuminuria and eGFR reduction represent independent risk factors for incident stage 653 CKD in T1DM patients. The simultaneous occurrence of reduced eGFR and albuminuria have a synergistic effect on renal function worsening

Advances in the treatment of prolactinomas

Prolactinomas account for approximately 40% of all pituitary adenomas and are an important cause of hypogonadism and infertility. The ultimate goal of therapy for prolactinomas is restoration or achievement of eugonadism through the normalization of hyperprolactinemia and control of tumor mass. Medical therapy with dopamine agonists is highly effective in the majority of cases and represents the mainstay of therapy. Recent data indicating successful withdrawal of these agents in a subset of patients challenge the previously held concept that medical therapy is a lifelong requirement. Complicated situations, such as those encountered in resistance to dopamine agonists, pregnancy, and giant or malignant prolactinomas, may require multimodal therapy involving surgery, radiotherapy, or both. Progress in elucidating the mechanisms underlying the pathogenesis of prolactinomas may enable future development of novel molecular therapies for treatment-resistant cases. This review provides a critical analysis of the efficacy and safety of the various modes of therapy available for the treatment of patients with prolactinomas with an emphasis on challenging situations, a discussion of the data regarding withdrawal of medical therapy, and a foreshadowing of novel approaches to therapy that may become available in the future

Case report : cystoscopic use of cyanoacrylate glue for bleeding during transurethral resection of bladder tumors

An 81-year-old woman with a history of nephroureterectomy and bladder cuff excision for grade 3 stage pT3N0M0 transitional-cell neoplasia presented with three bladder neoplasms. She also had symptomatic varicose veins in the lower extremities and received low-molecular-weight heparin. During transurethral resection of the tumors, deep ablation of the base of one lesion resulted in significant arterial bleeding that could not be controlled with normal endoscopic techniques. On the basis of previous positive experience, we endoscopicaly injected 3 mL of cyanoacrylate glue (Glubran 2) deeply (5 mm) into the tissue surrounding the bleeding site, making four injections with a device used for collagen injections. The area was irrigated with mannitol and sorbitol, and within a few seconds, there was complete remission of bleeding. We waited a further 90 seconds with the bladder distended to allow the substance to lose its adhesive properties so as to avoid having the bladder walls stick together. The catheter was removed on the third day, with resumption of micturition and clear urine. This appears to be the first case of cystoscopic use of cyanoacrylate glue to control bleeding

Robot-assisted radical prostatectomy in patients with a history of endoscopic treatment for benign hyperplasia of the prostate

Aim: The purpose of this study was to determine whether previous prostate surgery has an adverse effect on the perioperative, histopathological and functional outcomes of robot-assisted radical prostatectomy (RARP). Patients and Methods: We retrospectively identified 42 patients (Group 1) who had a history of endoscopic prostate surgery for the treatment of benign prostate hyperplasia (BPH). We performed one-to-one matching based upon ten variables (age, BMI, PSA, SHIM, AUA-SS, c stage, biopsy Gleason score, median lobe presence, pathological stage and the extent of nervesparing) using a propensity score matching algorithm to generate a control group (Group 2). The perioperative, histopathological and functional outcomes were compared between these groups. All the patients were evaluated for continence outcomes, while only those patients who had a preoperative SHIM score of 17 and had a nerve-sparing surgery (unilateral, bilateral or partial) were examined for potency outcomes. Results: The patient demographics after matching are listed in Table I. The perioperative histopathological and functional outcomes were evaluated. There was no statistically significant difference at any point in time for either continence or potency outcomes Conclusion: RARP is a safe and effective procedure in patients who have a history of endoscopic prostate surgery. Although the OR time is higher in these patients, the PSM rates, complications, histopathological and functional outcomes are comparable to those of patients who have no prior prostate surgery

Epidemiology of renal colic in a district general hospital

BACKGROUND/AIMS: To assess the incidence of renal colic and the results of emergency management. METHODS: During a 12 month period data of patients with symptoms of renal colic were collected. RESULTS: A total of 495 visits were registered. The M/F was 2.19. Mean age was higher in males (45.5+/-13.0 vs 42.5+/-15.5 years, P=0.025). Three patients were hospitalised for immediate urinary diversion due to anuria or sepsis. Fifty-three patients recovered without performing any pharmacological treatment. Analgesic treatment (mainly NSAID) was offered to 439 patients. After a 6 hour period 36 patients were admitted to the hospital owing to persistent pain. Pain was reduced in 403 patients (91.8%) who were offered outpatient renal ultrasound within 48 hours. Twenty-five patients (6.2%) required deferred hospitalisation. Follow up with renal ultrasound was obtained in 213. CONCLUSION: Renal colics accounted for 0.9% of ambulatory care visits to our emergency departments with an annual rate of 0.158 visits per 100 in the general population. NSAIDs were efficacious in the management of colic. Diagnostic work up was able to demonstrate the presence of a stone in 56% of the subjects presenting with renal colic whereas alternative diagnoses were demonstrated in 12%