Analysis of the finasteride treatment and its withdrawal in the rat hypothalamus and hippocampus at whole-transcriptome level.

PURPOSE: As reported in patients treated for androgenetic alopecia with finasteride (i.e., a blocker of the enzyme 5 alpha-reductase) and in an animal model, side effects affecting sexual, psychiatric, neurological, and physical domains, may occur during the treatment and persist with drug suspension. The etiopathogenesis of these side effects has been poorly explored. Therefore, we performed a genome-wide analysis of finasteride effects in the brain of adult male rat. METHODS: Animals were treated (i.e., for 20 days) with finasteride (1mg/rat/day). 24 h after the last treatment and 1 month after drug suspension, RNA sequencing analysis was performed in hypothalamus and hippocampus. Data were analyzed by differential expression analysis and Gene-Set Enrichment Analyses (GSEA). RESULTS: Data obtained after finasteride treatment showed that 186 genes (i.e., 171 up- and 15 downregulated) and 19 (i.e., 17 up- and 2 downregulated) were differentially expressed in the hypothalamus and hippocampus, respectively. Differential expression analysis at the drug withdrawal failed to identify dysregulated genes. Several gene-sets were enriched in these brain areas at both time points. CONCLUSION: Some of the genes reported to be differentially expressed (i.e., TTR, DIO2, CLDN1, CLDN2, SLC4A5, KCNE2, CROT, HCRT, MARCKSL1, VGF, IRF2BPL) and GSEA, suggest a potential link with specific side effects previously observed in patients and in the animal model, such as depression, anxiety, disturbance in memory and attention, and sleep disturbance. These data may provide an important background for future experiments aimed at confirming the pathological role of these genes.

Family Pediatrician and Public Health collaboration, an alliance to increase vaccination coverage: an experience with MenB vaccination in Italy.

Background: Invasive Meningococcal Disease is a severe disease mainly affecting infants and young children. Most infections are caused by serogroups A, B, C, W, X, and Y. In the last 10 years, serogroup B has been the main cause of Invasive Meningococcal Disease in Europe. Recent data resulting from an observational study conducted in Italy show a significant reduction in the number of Invasive Meningococcal Disease cases due to Neisseria meningitidis B after the introduction of vaccine 4CMenB. Thus, the Naples Team of Federation of Italian Primary Care Pediatricians and the Public Health Department started an active collaboration focused on vaccination process management (named "Progetto Via") with the aim of increasing Meningococcal B vaccination coverage. Study design: Source of data is the regional platform "GE.VA.". Every Primary care Pediatrician uses daily to record vaccination activity. This platform is integrated with data entered by operators of the District/Vaccination Center. Methods: Time: January 2019 - December 2019. The Federation of Italian Primary Care Pediatricians/Naples organized a meeting to identify six coordinators. The pediatricians could choose to counsel in their own offices and send children to the vaccination center or to counsel and vaccinate directly in their own clinics. Results: A total of 78 pediatricians took part in the project: 46 did only counseling and 32 did both counseling and vaccination in their medical clinic. Data obtained show an overall average vaccination coverage growth of about 13% in the first 4 months of the survey, and a further growth of about 11% in the following seven months, with a total growth in the entire period of 24%. The pediatricians' counseling is essential to recover non-compliant subjects, considering both the relationship of trust with the families and the visits already scheduled as an ideal moment for vaccinations' status check. Conclusions: The project highlights how an effective collaboration between family pediatricians and the Local Health Authority becomes valuable in getting closer to reach the Ministerial goal of 95%. Vaccination coverage increased significantly when family pediatricians supported the activity of vaccine centers in distress in many regional situations. The trust relationship, the hourly availability and the capillary network of family pediatricians' clinics were key elements for the success of this project and were also recognized by parents.

Laparoscopic and open treatment of hepatocellular carcinoma in patients with cirrhosis.

BACKGROUND: Few studies have compared survival and recurrence rates between laparoscopic and open liver resection for hepatocellular carcinoma in patients with cirrhosis. METHODS: A retrospective analysis of a prospectively maintained database of 179 liver resections performed for HCC in cirrhotic liver between 2000 and 2007 was performed. RESULTS: Fifty-four patients underwent a laparoscopic resection and 125 patients had open surgery. Histopathological features were largely comparable in the two groups. Tumours operated laparoscopically were smaller than those removed at open operation and laparoscopic liver resection was less extensive. Laparoscopic surgery had a lower morbidity rate than open surgery (19 versus 36.0 per cent; P = 0.020), whereas 30-day mortality was similar (2 versus 4.0 per cent; P = 0.615). After a median follow-up of 24 months, 1- and 3-year survival rates were 94 and 67 per cent in the laparoscopic group. Recurrence rates were similar after laparoscopic and open procedures (45 versus 52.5 per cent; P = 0.381), as was disease-free survival (P = 0.864). CONCLUSION: Laparoscopic resection of HCC in cirrhotic liver is feasible and safe in selected patients. Adequate long-term survival and recurrence is achieved compared with open surgery, when stratified for tumour characteristics known to be related to survival outcome.

Laparoscopic versus open liver resection for hepatocellular carcinoma in patients with histologically proven cirrhosis: short- and middle-term results.

BACKGROUND: Liver surgery, especially for cirrhotic patients, is one of the last areas of resistance to progress in laparoscopic surgery. This study compares the postoperative results and the 2-year patient outcomes between laparoscopic and open resection for hepatocellular carcinoma in patients with histologically proven cirrhosis. METHODS: From May 2000 to October 2004, 23 consecutive cirrhotic patients who underwent laparoscopic hepatectomy (LH) for HCC were compared in a retrospective analysis with a historic group of 23 patients who underwent open hepatectomy (OH). The two groups were well matched for age, gender, American Society of Anesthesiology (ASA) class, tumor location and size, type of liver resection, and severity of cirrhosis. The selection criteria for both groups specified a small (size < 5 cm), exophytic, or subcapsular tumor located in the left or peripheral right segments of the liver (II-VI segments, Couinaud); a well-compensated cirrhosis (Child-Pugh A); and an ASA score lower than 3. In the LH group, 15 subsegmentectomies, 3 segmentectomies, and 5 left lateral sectionectomies were performed, as compared with 12 subsegmentectomies, 5 segmentectomies, and 6 left lateral sectionectomies in the OH group. RESULTS: One patient in the LH group (4.3%) underwent conversion to laparotomy for inadequate exposition. The mean operative time was statistically longer for the LH group (LH, 148 min; OH, 125 min; p = 0.016), whereas blood transfusions (LH, 0%; OH, 17.3%; p = 0.036), Pringle maneuver (LH, 0%; OH, 21.73%; p = 0.017), mean hospital stay (LH, 8.3 days; OH, 12 days; p = 0.047), and postoperative complications (LH, 13%; OH, 47.8%; p = 0.010) were significantly greater in OH group. There was no statistically significant difference in mortality and 2-year survival rates between the two groups. CONCLUSION: This study shows that LH for HCC in properly selected cirrhotic patients results in fewer early postoperative complications and a shorter hospital stay than the traditional OH. The 2-year survival rate was the same for LH and OH.

Tetracycline-regulatable expression vectors tightly regulate in vitro gene expression of secreted proteins.

The regulation of gene expression by the tetracycline system has attracted a high level of interest in the recent past. However, expression of secreted proteins has not been evaluated precisely. In this study, we constructed two versions of a one-plasmid system containing the elements necessary for the regulation of gene expression. The regulatable elements and the selectable marker (Neor) were set up in two different configurations, pTRIN31 and pTRIN76. With these two regulatable versions, the levels of protein expression after transfection into the NIH/3T3 cell line were measured by insertion of three different genes encoding the secreted proteins (hGH, ApoE3, hGM-CSF). The maximum levels of gene expression obtained with the pTRIN76-derived plasmids were 100ng/24h/106 cells for hGH, 427ng/24h/106 cells for ApoE3 and 108ng/24h/106 cells for hGM-CSF. For the pTRIN31-derived plasmids the maximum levels were 2.7ng/24h/106 cells for hGH and 47ng/24h/106 for ApoE3. Both plasmids give rise to an expression of the transfected gene that can be tightly regulated by three different molecules: tetracycline, minocycline and doxycycline. The levels of the secreted proteins are below the detectable level when the reporter genes are repressed. This repression is reversible within 48h after the regulator has been removed from the medium.

Lack of diet-induced thermogenesis following lesions of paraventricular nucleus in rats.

The effects of electrolytic lesions in the hypothalamus paraventricular nucleus were studied in adult male and female Sprague-Dawley rats, fed different diets, consisting of either palatable human food plus chow (cafeteria diet) or chow alone. The results showed that both cafeteria diet and lesions induced an increase in energy intake and weight gain in rats of both sexes. Oxygen consumption rate and colonic temperature were significantly decreased by lesions, while cafeteria diet increased the same parameters only in intact animals. The lesion decreased weight, protein and DNA, and temperature of brown adipose tissue, while cafeteria diet increased the values considered in brown adipose tissue of sham-injured rats, but not in lesioned animals. The response to norepinephrine administration was significantly greater in intact rats and those fed cafeteria diet. The results suggest that the larger body weight gain observed in lesioned rats, particularly evident in rats fed cafeteria diet, is partly due to the disappearance of diet-induced thermogenesis that depends on the reduced mass and functional activity of brown adipose tissue.

Postprandial thermogenesis and conditioned taste aversion or preference.

Postprandial thermogenesis is under the control of the autonomic nervous system and alimentary conditioned stimuli change sympathetic and parasympathetic activity. Here we studied the effect of conditioned taste aversion on postprandial thermogenesis in rats. Two groups of animals were used, rats of the first group were controls, these were placed on a standard diet and, for some days, on two other different diets: one thiamine-free and the other thiamine-rich. Each diet had a different taste. The treated animals belonged to the second group, these were fed with the same three diets but for different lengths of times: thiamine-free diet for the first 5 wk afterwards, with thiamine-rich diet for 3 wk, and finally with laboratory standard diet for a few days. After a preference test with the three familiar diets, oxygen consumption rate and brown adipose tissue temperature were evaluated three times in both groups after ingestion of a test meal, each time with one of the three different diets. The preference test was unvaried for the three different familiar foods in controls, while the treated animals showed a lower preference for thiamine-free food than for the other two. Treated rats had a significantly higher increase in O2 consumption rate than controls. In this group intake of thiamine-free food induced a significantly lower increase in O2 consumption than the other two. The increase in brown adipose tissue temperature was also higher in treated than in control animals but in treated rats this was lower after intake of thiamine-free food than after the intake of the other two.(ABSTRACT TRUNCATED AT 250 WORDS)

Is 3000 S Hospal a suitable dialyzer to shorten dialysis time schedule? A cooperative study on biofiltration.

The Biofilter 3000 S Hospal may combine higher convective clearance rates (Cc) with usual diffusive clearance rates (Cd) (i.e. similar to Cuprophan dialyzers), giving a higher total clearance rate (Ct) of small and middle molecules. Use of the Biofilter has been suggested to shorten dialysis time schedules. This study was carried out in 8 patients on RDT 3 times weekly, by cuprophan filter and acetate dialysis. The patients were shifted to dialysis with 3000 S guided by two principles: to shorten dialysis time by 1 hour per session, and to reinfuse 6 liters of bicarbonate-saline solution (40 mEq/l) per single dialysis. Besides the usual clinical and laboratory controls, in three patients clearance studies were carried out during four different dialysis sessions: Ct, Cc and Cd of urea K+, creatinine, uric acid and phosphate were measured. No change was observed in the main clinical and laboratory parameters after 3-5 months (average 3.9) of treatment with Biofilter 3000 S; in addition, serum alkaline phosphatase concentration decreased progressively. Clearance results, however, indicate that the expected high values of Ct do not occur, because Cd decreases as Cc is increased. A primary goal of research in hemodialysis is to reduce the average time of treatment while ensuring simultaneously "physiological" dialysis. A possible approach to this problem is to use dialyzers with highly permeable and biocompatible membranes such as the "biofilter" 3000 S Hospal.(ABSTRACT TRUNCATED AT 250 WORDS)

Molecular aspects of resistance to biological and non-biological drugs and strategies to overcome resistance in colorectal cancer.

A number of successful systemic therapies are available for the treatment of disseminated cancers. However, tumor response is often transient, and therapy frequently fails due to emergence of resistant populations. The latter reflects the temporal and spatial heterogeneity of the tumor microenvironment as well as the evolutionary capacity of cancer phenotypes to adapt to therapeutic perturbations. Resistance to either chemotherapy and targeted agents limits the effectiveness of current cancer therapies, including those used to treat metastatic colorectal cancer (mCRC) which is one of the leading causes of cancer-related death worldwide. Resistance to therapeutic drugs can be already present at diagnosis or it can develop after treatment. These two forms of resistance are respectively called intrinsic and acquired. The identification of mechanisms of drug resistance may highlight new biomarkers useful to predict the clinical outcome or the likely responsiveness to pharmacological treatment of those metastatic CRC patients who cannot benefit from current therapeutic regimen. Moreover, the recognition of panels of biomarkers may suggest new strategies to overcome resistance by rational drug design and combination treatment. In this review, we describe molecular mechanisms of resistance to chemotherapies and targeted agents that may be relevant to colorectal cancer and the possible strategies to overcome the resistance.

Ultrasonically activated device for parenchymal division during open hepatectomy.

BACKGROUND: The use of new technological devices has gained popularity and has been proposed to improve the safety of liver resection. This study was designed to evaluate the usefulness of the ultrasonically activated device (USAD) during open liver resection. MATERIALS AND METHODS: Indication for surgery, type of resection, need to perform a Pringle manoeuvre, operation time, blood loss, number of blood transfusions, morbidity and mortality rate were analyzed in 60 patients undergoing a formal open liver resection by means of USAD. RESULTS: The overall mean operation time was 172 minutes (range 120-255 min); an intermittent warm ischemia was applied in 9 cases (15%). The overall mean blood loss was 410 mL (median 400 mL, range 50-950 ml). A median of one blood transfusion was administered in six patients (10%). The mean hospital stay was 10.2 days (median 11, range 8-16). The overall morbidity rate was 20% (12 out of 60 patients). No in-hospital mortality was recorded. By subdividing the patients according to the presence or absence of cirrhosis no statistical significant differences were found between the two subgroups in all peri-and postoperative outcomes. CONCLUSIONS: In conclusion, though there is a lack of data based on well conducted controlled studies and further on a greater number of patients are needed, the utilization of USAD may help to minimize blood loss during liver resection regardless of the condition of the liver, even in case of cirrhosis.

General theory of critical periods and development of obesity.

The general systems theory (GST), the general theory of organization (GTO), and the general theory of critical periods (GTCP) have been applied to some nutritional problems. This theoretical approach seems to be in good agreement with most of the data of the literature and with the personal experience, pointing at the possibility to use a simple general model to interpret the complex problem of obesity.

[Effects of oral glucose load on blood lipids and on platelet function]. / Effetto del carico orale di glucosio sui lipidi ematici e sulla funzione piastrinica.

Oral glucose tolerance test (OGTT) has been performed on 12 healthy men. Plasma glucose, cholesterol, triglycerides, total lipids, NEFA, and number, adhesiveness and aggregating of platelets have been measured before and during OGTT (at 30, 60, 90, 120, 150 mins). Results demonstrate that OGTT decreases during the 1st hour, increases during the 2nd hour, and brings again to the initial value the platelet aggregation during the 3rd hour. The biphasic trend of platelet aggregation, in comparison with the monophasic OGTT curve, allows us to hypothesize some relationships between platelet aggregation and not only blood glucose concentration but also the activity of some hormones controlling both blood sugar level and platelet function.

A novel endothelial cell-based gene therapy platform for the in vivo delivery of apolipoprotein E.

A major focus in gene therapy has been the use of recombinant viruses to deliver genes in vivo. Although this approach shows much promise, there are many safety concerns associated with the use of viral materials in the treatment of human diseases. Our alternative cell-based gene therapy approach utilizes endothelial cells (Pro 175) isolated from the murine embryonic yolk sac. These endothelial cells were evaluated for their potential use in gene therapy as a gene delivery platform. As a test model, we used these cells to deliver apolipoprotein E (apoE) in the murine apoE knockout atherosclerosis model. The lack of apoE protein in these animals results in high levels of serum cholesterol and formation of severe aortic plaques and lesions at a young age. After transplantation of the apoE secreting Pro 175 endothelial cells into apoE-deficient mice, serum cholesterol levels were measured at 2 week intervals. During the 3 months after the initiation of these experiments, levels of cholesterol in the animals having received the apoE secreting endothelial cells were statistically lower compared with the levels of age-matched controls having received non-secreting endothelial cells. Concomitant with cholesterol reduction, atherosclerotic aortic plaques were noticeably reduced in the experimental apoE+ animals. These results highlight the potential of these unique endothelial cells as an efficient delivery platform for somatic gene therapy.