An alternative, arginase-independent pathway for arginine metabolism in Kluyveromyces lactis involves guanidinobutyrase as a key enzyme.

Most available knowledge on fungal arginine metabolism is derived from studies on Saccharomyces cerevisiae, in which arginine catabolism is initiated by releasing urea via the arginase reaction. Orthologues of the S. cerevisiae genes encoding the first three enzymes in the arginase pathway were cloned from Kluyveromyces lactis and shown to functionally complement the corresponding deletion in S. cerevisiae. Surprisingly, deletion of the single K. lactis arginase gene KlCAR1 did not completely abolish growth on arginine as nitrogen source. Growth rate of the deletion mutant strongly increased during serial transfer in shake-flask cultures. A combination of RNAseq-based transcriptome analysis and (13)C-(15)N-based flux analysis was used to elucidate the arginase-independent pathway. Isotopic (13)C(15)N-enrichment in γ-aminobutyrate revealed succinate as the entry point in the TCA cycle of the alternative pathway. Transcript analysis combined with enzyme activity measurements indicated increased expression in the Klcar1Δ mutant of a guanidinobutyrase (EC.3.5.3.7), a key enzyme in a new pathway for arginine degradation. Expression of the K. lactis KLLA0F27995g (renamed KlGBU1) encoding guanidinobutyrase enabled S. cerevisiae to use guanidinobutyrate as sole nitrogen source and its deletion in K. lactis almost completely abolish growth on this nitrogen source. Phylogenetic analysis suggests that this enzyme activity is widespread in fungi.

Occupational Therapy's efficacy in children with Asperger's syndrome: a systematic review of randomized controlled trials.

This systematic review of randomized controlled trials (RCTs) aims at evaluating the efficacy of Occupational therapy (OT) interventions in Asperger's syndrome (AS) pediatric patients. We conducted a systematic review according to PRISMA guidelines. The screening of the literature was carried out on PUBMED, SCOPUS, WEB OF SCIENCE and OT SEEKER databases, TO December 2018. We selected three RCTs having the common objective to evaluate whether children with AS can improve their social skills thanks to OT treatments. The interventions targeted by the three selected studies were: LEGO therapy, Social Use of Language Programme, Let's Face It! software and Parent Training combined with the administration of risperidone. All the studies showed that the intervention groups improved their overall social ability.This review shows that OT interventions can help concretely AS children in overcoming their social issues. Nevertheless, more RCTs are needed to better understand the OT's benefits and limitations in AS patients.

The interaction of human dendritic cells with yeast and germ-tube forms of Candida albicans leads to efficient fungal processing, dendritic cell maturation, and acquisition of a Th1 response-promoting function.

T helper cell type 1 (Th1) cell-mediated immunity plays a critical role in protection against the opportunistic pathogen Candida albicans. Virulence of the fungus is closely associated with its ability to form germ-tubes (GT), the early phase of the dimorphic transition from the commensal yeast (Y) to the more invasive hyphal (H) form. In this study, we examined the functional outcome of the interaction of Y or GT forms with human dendritic cells (DCs), professional antigen-presenting cells, which are pivotal for initiation and modulation of T cell responses. DCs phagocytosed and killed Y and GT cells with a comparable efficiency, becoming able to trigger strong proliferative responses by Candida-specific, autologous T cell clones. Both fungal forms induced DC maturation, as indicated by up-regulation of CD83, CD80, CD86, CD40, and major histocompatibility complex classes I and II surface antigens. Chemokine receptors were also modulated in Candida-DCs, which showed increased CCR7/CXCR4 and decreased CCR5 expression. Y- and GT-activated DCs differed in the pattern of cytokine expression. In particular, GT cells, in common with fully differentiated H cells, induced significantly more elevated levels of interleukin (IL)-10 than Y cells. Nevertheless, Y-, GT-, or H-pulsed DCs secreted comparable amounts of IL-12p70. In addition, irrespective of the fungal form triggering DC activation, Candida-DCs acquired the ability to prime naive T lymphocytes with a defined Th1 phenotype. Overall, our findings highlight the induction of substantially similar functional patterns in human DCs encountering the different forms of growth of C. albicans, both seemingly activating the Th1-type immunity which is characteristic of the healthy human subjects, naturally immunized and protected against the fungus.

Expression and function of neurotrophins and their receptors in cultured human keratinocytes.

Whereas nerve growth factor has been extensively studied in human keratinocytes, little is known on the role of other members of the neurotrophin family. We investigated the expression and function of neurotrophins and neurotrophin receptors in cultured human keratinocytes. We demonstrated by reverse transcription-polymerase chain reaction that keratinocytes synthesize neurotrophin-3, brain-derived neurotrophic factor, and neurotrophin-4/5. These cells also express tyrosinase kinase A and C, the nerve growth factor and neuro-trophin-3 high-affinity receptors, respectively. On the other hand, only the truncated extracellular isoform of tyrosinase kinase B, the high-affinity brain-derived neurotrophic factor and neurotrophin-4/5 receptor, is detected in keratinocytes. Moreover, neurotrophin-3, brain-derived neurotrophic factor, and neurotrophin-4/5 proteins are secreted by human keratinocytes at low levels. Keratinocyte stem cells synthesize the highest amounts of nerve growth factor, while they secrete higher levels of nerve growth factor as compared with transit amplifying cells. Neurotrophin-3 stimulates keratinocyte proliferation, where brain-derived neurotrophic factor or neurotrophin-4/5 does not exert any effect on keratinocyte proliferation. Addition of neurotrophin-3 slightly upregulates the secretion of nerve growth factor, whereas nerve growth factor strongly augments neurotrophin-3 release. Ultraviolet B irradiation downregulates nerve growth factor, whereas it augments neurotrophin-3 and neurotrophin-4/5 protein levels. Ultraviolet A irradiation increases the level of neurotrophin-3, whereas it does not exert any effect on the other neurotrophins. Finally, neurotrophins other than nerve growth factor fail to protect human keratinocytes from ultraviolet B-induced apoptosis. This work delineates a functional neurotrophin network, which may contribute to epidermal homeostasis.

Carnitine deficiency induced during hemodialysis and hyperlipidemia: effect of replacement therapy.

Plasma carnitine levels were studied in 14 uremic patients before, during, and after hemodialysis. The predialysis plasma carnitine levels were normal but fell during dialysis (half-life of 3.6 h). Plasma carnitine levels rose quickly in the first 6 h after dialysis, after which time the rise was more gradual. Muscle carnitine was significantly reduced in the dialyzed patients (p less than 0.005) compared with controls. In four patients lipid droplets were observed in muscle. Ten patients on maintenance hemodialysis exhibited plasma hyperlipidemia and low muscle carnitine. These individuals were given DL-carnitine (50 mg/kg body weight) intravenously after each dialysis. At the end of a 2-month carnitine treatment, plasma triglyceride levels were found to be reduced (p less than 0.001) and muscle carnitine content significantly increased (p less than 0.005). These findings suggest that carnitine may be useful in treatment of hypertriglyceridemia and muscle carnitine deficiency states induced during maintenance hemodialysis.

A background-free assay for cells forming antibody to glucose oxidase coupled with an enzyme-linked immunosorbent assay (ELISA) to quantitate antibody secretion.

We describe an immunocytochemical assay for cells forming antibody to glucose oxidase (GO). The method is specific in that only cells containing intracytoplasmic antibody capable of binding the immunogen (GO binding cells; GOBC) are stained. The method is sensitive because there is no GO activity in mammalian tissues. This lack of background readily permits detection of one GOBC among 10(6) nucleated lymphohemopoietic cells. The technique is reliable because purified chemicals are used. Although it is not possible to determine the Ig class of antibody formed by an individual cell, as can be done with the hemolytic plaque assay, the amount and class of secreted antibody to GO can be quantitated by an indirect enzyme-linked immunosorbent assay (ELISA), which is also described. GO is immunogenic and stimulates the formation of large numbers of GOBC in the popliteal lymph nodes after injection with adjuvant into the footpads of mice, but 1-mg doses injected IV or IP are lethal because of its enzymatic activity, which causes hypoglycemia and methemoglobinemia.

The microsurgical drainage of the varicocele.

A new technique for surgical treatment of primary and recurrent varicocele is proposed. The technique consists of direct anastomosis of two or three dilated veins of the pampiniform plexus with the great saphenous vein. A connection between a high venous pressure region and a lower pressure region is established. Twenty-five patients were operated on, 7 with recurrent varicocele and 18 with primary varicocele. The immediate and long-term results (6 year follow-up) have always been good.

Beneficial effects of L-carnitine in dialysis patients with impaired left ventricular function: an observational study.

BACKGROUND: Recent studies have shown that L-carnitine may improve clinical status and reduce the need for erythropoietin in dialysis patients with cardiovascular diseases. In this observational study, we investigated whether the addition of L-carnitine to conventional therapy might improve cardiac function (as assessed by M-mode and two-dimensional echocardiography) and clinical status in dialysis patients with left ventricular dysfunction. METHODS: Eleven dialysis patients with reduced left ventricular function (EF < 45%) were treated with L-carnitine for 8 months. Two-dimensional (2-D) echocardiography was performed at baseline and every 2 months up to the end of the treatment period. The dosage of erythropoietin was also monitored during the study and the patients' clinical status was assessed by a questionnaire. RESULTS: Carnitine increased mean LV ejection fraction from 32.0% to 41.8% (p < 0.05 vs baseline). There was also a slight reduction of erythropoietin dosage and an improvement of clinical status. CONCLUSIONS: Eight months' therapy with carnitine appears to improve LV function and clinical status in dialysis patients with impaired LVF.

Renal function during calcitriol therapy in chronic renal failure.

Some authors have reported that vitamin D treatment worsens renal function. We studied 10 patients with stable renal function, who were given calcitriol (0.5 micrograms/day) for a 4-month period. Creatinine and inulin clearance were performed at the beginning and at the end of the treatment. Although serum creatinine increased and creatinine clearance decreased, inulin clearance did not show significant variations. Furthermore, serum creatinine fell to the baseline value within 60 days after discontinuation of vitamin D therapy. The increased serum creatinine may be explained by an augmented release from muscular tissue, probably due to the improvement of uremic myopathy induced by calcitriol.

Induction of the multidrug-transporter P-glycoprotein by 3'-azido-3'-deoxythymidine (AZT) treatment in tumor cell lines.

A major form of multidrug resistance, which represents a serious obstacle to the success of chemotherapy, is caused by the over-expression of MDR-1 gene encoded P-glycoprotein. The present investigation was aimed to determine whether AZT, a cytostatic agent that interferes with the human immunodeficiency virus replication, is able to induce MDR-1 expression in tumor cells. After a short term exposure of human lymphoblastoid cells to AZT MDR-1 P-glycoprotein was found in the treated cells. This ATP-dependent drug-efflux pump interferred with cytotoxic efficacy of anticancer drugs such as vinblastine. This phenomenon should be carefully considered during anti-viral and anti-tumoral combined chemotherapies in AIDS patients.

Biofiltration vs. bicarbonate dialysis: influence on plasma volume changes and extravascular fluid mobilization.

The present study compares the effects of bicarbonate hemodialysis (Bic. HD) and biofiltration (BF), a new hemodiafiltration technique, on plasma volume (PV) changes and extravascular fluid mobilization (Vfm). Ten uremic patients underwent one experimental session of Bic. HD and, one week later, one of BF, both on the second dialysis of the week. Net ultrafiltration rate was limited to 700 ml/min. At the start of each session, whole blood volume (WBV), PV and red cell volume (RCV) were determined using 5 mu Ci of radioiodinated serum albumin (RISA). PV and Vfm were calculated at hourly intervals using a serial hematocrit method. On Bic. HD, PV increased at 60 min. then decreased at 120 and 180 min., with efficient Vfm only during the first hour. On BF, PV increased throughout treatment, with greater Vfm. It would appear that PV is better preserved in BF, on account of more efficient Vfm.

Telemedicine technology and applications for home hemodialysis.

Home hemodialysis (HD) for the treatment of patients with end-stage renal disease (ESRD) was first put into practice about 30 years ago. In this paper we describe the application of telematics monitoring services (TMS) for supporting patients who need home or satellite HD (SHD). For the clinical trials two modified HD machines were located in the renal unit and a central control station (UNIX workstation with multimedia PC-terminal) was located in another room of the hospital. Bi-directional communication between modified HD machines and central control station was managed via ISDN (Integrated Services Digital Network) links. Using these HD-machines 150 HD sessions were performed in nine patients over a period of five months. This system enabled on-line remote supervision of the HD machine-related functions (air in the blood, leak of blood, low conductivity etc.) and the clinical condition of patients through measurement of blood pressure (BP), pulse rate, PO2 (pulse oxymetry) and electrocardiogram (ECG) from the central control station (CCS). The user checked the type of alarm/warning, its appearance on HD machines and multimedia terminal units (MTU), the action of the protective system and the appearance of consultative messages from CCS on the remote terminal unit RTU. According to the data collected, the disturbances of HD machine function were visible and audible in the CCS and the user messages were always observed on the RTU. No unusual dialysis-associated complications were observed, all data and alarms/warnings were transmitted correctly and patients had adequate HD treatment.

Experimental lymphatico-venous shunt in the rat: pressure controls and long-term patency.

In a series of rats which underwent end-to-side lymphaticovenous shunts, between precaval lymphatic collector and infrarenal cava vein, pressure values in lymphatic and venous systems and long-term patency of the anastomosis have been evaluated. Lymphatic pressure has always been higher than venous pressure, providing a good physiological basis for the functioning of the shunt. Short-term patency (15 days) was 100% and long-term patency (90 days) 80% in all the controlled animals. The results obtained in this experimental model seem to support the efficacy of this procedure also in its clinical use.

[Renal transplantation. 1st year's experience at a new center]. / Trapianto renale. Primo anno di esperienza di un nuovo centro.

The initial activity in the field of organ transplantation always brings some organizational and medical difficulties. We describe the first year experience in the field of kidney transplantation at the Transplantation Unit of Padua University. Sixteen kidney transplants from cadaver donors have been performed. A double drug therapy (cyclosporine and steroids) as immunosuppression was used in 9 patients and a triple drug protocol (cyclosporine, steroids, azathioprine) was used in 7. Two patients also received monoclonal antibodies because of steroid resistant rejection. The technical problems we observed include one case of ureteral duplication corrected with uretero-ureteral anastomosis and one substenosis of an ureteral cystoanastomosis corrected with endoscopic dilatation. Among the complications related to immunosuppressive therapy we had one case of severe cyclosporine nephrotoxicity. Two patients died after few months with a functional graft because of pulmonary embolism and because of severe hepatitis complicated by pancreatitis. At an average of six months follow-up all the remaining patients have a good functioning graft.

[Study of the arrhythmia in patients with chronic uremia in periodic hemodialysis with Holter dynamic electrocardiography]. / Studio delle aritmie negli uremici cronici in emodialisi periodica mediante elettrocardiografia dinamica secondo Holter.

For the purpose of evaluating cause, frequency, type and seriousness of arrhythmias in dialysis patients, 14 chronic uremics, 8 on bicarbonate-dialysis, 6 on acetate-dialysis underwent a basal ECG, echocardiography and a Holter dynamic electrocardiography (ECGD) for a duration of 96 hours. Before and after dialysis PAO, body weight, serum electrolytes and arterial pH were controlled. In 11 patients (78%) supraventricular and ventricular arrhythmias were discovered of equal frequency and seriousness both in the inter and intra dialytic phase, even if more frequent in ventricular hypertrophic patients (IVSn) the complex ventricular arrhythmias (Lown greater than 2). The seriousness and frequency of ventricular and supraventricular arrhythmias in the dialytic phase did not seem to depend either on the type of tampon or on the presence or absence of cardiopathies while in the inter-dialytic phase the seriousness of ventricular arrhythmias seems to depend upon the presence of cardiopathies. The Authors conclude that the pathogenesis of arrhythmias in uremic patients on dialysis is multifactorial and that their elevated incidence makes the use of a Holter in these patients recommendable.

Evaluating dental office ergonomic. Risk factors and hazards.

Concerns about ergonomic risk factors have been growing among health care and other workers. The authors undertook a study to determine the presence of certain ergonomic risk factors in typical tasks performed by dentists and dental hygienists. They further investigated--by means of electromyography and goniometry--the force, frequency and duration of the task. The study suggests that when a task's duration, force and frequency are accounted for, scaling, polishing, flossing and probing activities do not represent exposure to high ergonomic risk.

Toriello-Carey syndrome: report of a new case.

Toriello-Carey syndrome is a malformation syndrome characterized by cranio-facial defects, visceral defects, congenital heart defects, hypotonia, and postnatal growth retardation. To our knowledge, only five patients with this condition have been reported. We describe another case of the Toriello-Carey syndrome extending the phenotypic manifestations of the syndrome.

Microsurgical treatment of lymphedemas of the limbs.

Lymphaticovenous anastomosis for surgical treatment of lymphedema of the limbs has been performed for several years with good results. The authors report on their experience, gained since 1976, after having operated on primary or secondary lymphedemas (five upper and 15 lower limbs). Lymphaticovenous anastomosis was performed according to Degni's technique using a special needle by which lymphatic vessels are inserted in the vein and secured to its wall. Eighty-three per cent of the subjective results were good, and the percentage of mean volumetric reduction in the operated limbs was 43% in 16 controlled cases. The postoperative course of secondary lymphedemas of the upper limbs was sometimes complicated by sudden lymphangitis attacks. In dealing with primary lymphedemas, the best results were obtained when all the operated limbs had great volumetric reduction, allowing the patients to resume their normal activities. The authors conclude that, after a long period of preoperative anti-inflammatory and anti-microbial therapy and regardless of lymphographic patterns, it is always worthwhile to perform a surgical exploration of the crural lymph nodes.

[Microsurgery and associated therapies for the treatment of lymphedema of the limbs]. / La microchirurgie et les thérapies associées pour le traitement des lymphoedèmes des membres.

Between 1967 and 1986, 170 cases of primary or secondary lymphedema of upper or lower limbs were operated upon using the indirect lymphaticovenous anastomosis technique proposed by Degni. For the lymphedema of upper limbs secondary to axillary lymphadenectomy for breast cancer, satisfactory results were obtained in 40% of cases, the figure rising to 72 and 74% for primary and secondary lymphedema of lower limbs respectively. When surgery is unsuccessful or contraindicated a new therapeutic method is proposed: thermotherapy as developed by Pr. Chang's team in Shanghai based on an ancient traditional Chinese medical method. With the collaboration of a biomedical engineer, a new microwave oven has been constructed and tested for the treatment of lymphedema, and its characteristics are described. Treatment is ambulatory, 45 minutes per day, 5 days per week over 3 weeks. It is very important that patients wear an elastic contention stocking (40 mmHg) between sessions. A total of 55 patients have been treated in this way, most (45) failing to improve after other medical or surgical treatment. All cases treated were of severe lymphedema and subjective and objective (ultrasonography) improvement was noted in 51%, the other 49% obtaining only subjective relief. According to Chinese authors results are maintained after 10 years. Follow up of the present series has been a maximum of one year and longer surveillance is necessary to assess long-term efficacy.

Definition of essential amino acid residues in the recognition of a peptide by a mouse monoclonal antibody.

A mouse monoclonal antibody reacting in ELISA with a synthetic peptide representing a linear amino acid stretch of the protein antigen was tested on all overlapping 5-mer to 9-mer fragments of the peptide, as prepared by multi-pin synthesis. Analysis of the binding data suggests that several residues in the peptide might be relatively unrelevant for recognition, while few others seem to play a critical role as key residues. On the basis of such observations, we attempted to reconstruct an alternative essential epitope by introducing multiple amino acid substitutions in the 9-mer peptide exhibiting the best binding activity, and then tested its ability to be recognized by the monoclonal antibody.