Hepatocyte-conditioned medium sustains endothelial differentiation of human hematopoietic-endothelial progenitors

Liver neo-angiogenesis plays a fundamental role in physiological and pathological processes such as regeneration, cirrhosis, autoimmune hepatitis, and alcoholic liver disease. How liver parenchymal cells influence angiogenesis is largely unknown. We studied the influence of soluble factors released by hepatocytes on hematopoietic and endothelial cell differentiation. Human CD34+ cells cultured for several weeks in a hepatocyte-conditioned medium gradually decrease the expression of CD34 and CD133 markers (i.e. after 4 weeks from 85% and 69%, respectively, to 6% and 3%, respectively), whereas expression of CD144 and CD14 cell markers increased (from 2% and 8%, respectively, to 54% and 55%, respectively). The cells' capacity to form hematopoietic colonies in methylcellulose declined with time, whereas they acquired endothelial morphology, expressed endothelial markers, and incorporated into newly forming vascular structures both in vitro and in vivo. Cultured single CD34+ cells formed colonies expressing both hematopoietic (CD45+) and endothelial (CD144+) markers, suggesting they constitute a bona fide hemangioblast population. CONCLUSION: This system allowed subsequent stages of differentiation of hematopoietic cells to endothelial cells to be defined, underlining the strict interrelationship between endothelial and hematopoietic cells in a hepatocyte environment.     

Diagnostic value of HSP70, glypican 3, and glutamine synthetase in hepatocellular nodules in cirrhosis

Hepatocellular nodules in cirrhosis include regenerative (large regenerative, LRN) and dysplastic (low and high grade, LGDN and HGDN) nodules, early and grade 1 HCC (eHCC-G1), and overt HCC. The differential diagnosis may be particularly difficult when lesions such as HGDN and eHCC-G1 are involved. We investigated the diagnostic yield of a panel of 3 putative markers of hepatocellular malignancy such as HSP70, glypican 3 (GPC3), and glutamine synthetase (GS). We selected 52 surgically removed nonmalignant nodules (15 LRNs, 15 LGDNs, 22 HGDNs) and 53 HCCs (10 early, 22 grade 1, and 21 grade 2-3) and immunostained them for HSP70, GPC3, and GS. The sensitivity and specificity of the individual markers for the detection of eHCC-G1 were 59% and 86% for GS, 69% and 91% for GPC3, and 78% and 95% for HSP70. We identified 2 main phenotypes: (1) all negative, seen in 100% LRN and LGDN, 73% HGDN and 3% eHCC-G1; (2) all positive, a feature detected in less than half the eHCC-G1. Using a 3-marker panel, when at least 2 of them, regardless which, were positive, the sensitivity and specificity for the detection of eHCC-G1 were respectively 72% and 100%; the most sensitive combination was HSP70+/GPC3+ (59%) when a 2-marker panel was used. CONCLUSION: The adopted panel of 3 markers is very helpful in distinguishing eHCC-G1 from dysplastic nodules arising in cirrhosis.     

Cardiovascular complications of obesity in adolescents

Obesity is an increasingly important worldwide health problem, representing the major risk factor for coronary heart disease. The increase in the prevalence of obesity, particularly among younger age groups, is likely to have long-term implications for cardiovascular disease (CVD) in the years to come, especially at a young age. Obesity plays a central role in the insulin resistance (IR) syndrome and increases the risk of atherosclerotic CVD. The present review will examine the relationships among cardiovascular risk (CVR) factors during the childhood-adolescence-adulthood transition. In fact, the relation between obesity, in particular visceral obesity and CVD, appears to develop at a relatively young age. The foremost physical consequence of obesity is atherosclerotic CVD, and an intriguing example of obesity-related cardiovascular complications affecting young women is the polycystic ovary syndrome (PCOS).     

Use of western blot to study Microsporum canis antigenic proteins in canine dermatophytosis

Western blotting was used to describe the Microsporum canis proteins with antigenic activity in dogs with dermatophytosis. Electrophoretic separation of whole fungal strain extract cultured from a cat was performed under denaturing conditions. The proteins were blotted onto nitrocellulose and probed with sera collected from 22 dogs with dermatophytosis (18 M. canis, 3 M. gypseum, 1 Trichophyton mentagrophytes; group A), 20 dogs with skin diseases other than dermatophytosis, and 22 dogs with no clinical cutaneous signs (group B, n = 42). Nine principal IgG‐binding proteins with apparent molecular weights of 180, 144, 130, 120, 102, 96, 80, 68, and 48 kD were visualised on group A blots. For these proteins, serological cross‐reactivity with different strains of M. canis may be indirectly confirmed, whereas additional proteins were found to react with sera from individual dogs. The proteins visualised in this study may represent diagnostic markers of dermatophyte infection. The proteins should be further evaluated for their role in the cellular immune response of dogs with dermatophytosis.     

Life-threatening serotonin toxicity due to a citalopram-fluconazole drug interaction: case reports and discussion

OBJECTIVE: To discuss two cases of life-threatening serotonin toxicity due to a drug interaction between citalopram and fluconazole and to review the pertinent literature. METHODS: A Medline search without date limitation was conducted using the terms serotonin syndrome, serotonin toxicity, fluconazole and citalopram. RESULTS AND DISCUSSION: Fluconazole inhibits CYP2C19. Citalopram is a substrate for 2C19 and inhibition of its metabolism may result in serotonin toxicity. Serotonin toxicity in oncology patients may not present with the classic constellation of signs typically described in the literature. Delirium may be the only presenting feature. Current level of evidence for treatment of serotonin toxicity is level 4 or 5 (case series and expert opinion). Nevertheless, there is a strong theoretical basis for treating serotonin toxicity in medical patients with a 5H(2A) blocker such as cyproheptadine. CONCLUSIONS: Consultation-liaison psychiatrists and oncologists should be aware of this preventable and underrecognized interaction. Citalopram should be stopped or substituted prior to the concurrent administration of fluconazole, and in the event of toxicity, treatment with cyproheptadine has a favorable risk-benefit ratio despite a lack of randomized controlled data to support its use.     

Usefulness of atherogenic dyslipidemia for predicting cardiovascular risk in patients with angiographically defined coronary artery disease

The identification of factors contributing to residual cardiovascular risk is important to improve the management of patients with established coronary artery disease (CAD). This study was conducted to assess the predictive value of atherogenic dyslipidemia (defined as high triglycerides and low high-density lipoprotein [HDL] cholesterol) for long-term outcomes in patients with CAD. In 284 patients (238 men, 46 women; mean age at baseline 59.2 +/- 8.9 years) with coronary stenosis (>50% in > or =1 vessel), the presence of atherogenic dyslipidemia was prospectively associated with the incidence of major adverse cardiovascular events (MACEs) during a median follow-up of 7.8 years. MACEs were defined as cardiovascular death, nonfatal myocardial infarction, nonfatal stroke, the recurrence of angina, and revascularization procedures. MACEs were observed in 111 (39.1%) patients with CAD. MACEs occurred more frequently in patients with atherogenic dyslipidemia (50.9%) than in those with isolated low HDL cholesterol or high triglycerides (33.0%) or with normal HDL cholesterol and triglyceride concentrations (29.2%) (p <0.01 for trend). Kaplan-Meier survival analysis showed a decrease in event-free survival in patients with compared with those without atherogenic dyslipidemia (log-rank p = 0.006). Patients with atherogenic dyslipidemia presented with increased plasma concentrations of remnants, denser low-density lipoprotein, more atherogenic HDL particles, and insulin-resistant status. After adjustment for potential confounding variables, the magnitude of increased risk associated with atherogenic dyslipidemia was 1.58 (95% confidence interval 1.12 to 2.21, p = 0.008). In conclusion, these data provide evidence that atherogenic dyslipidemia is an independent predictor of cardiovascular risk in patients with CAD, even stronger than isolated high triglycerides or low HDL cholesterol.     

Cognitive-behavioural stimulation protocol for severely brain-damaged patients in the post-acute stage in developmental age

Purpose. To present a cognitive-behavioural stimulation (CBS) protocol designed to help severely damaged patients in the early post-acute stage by describing the underlying methodology and assessing its efficacy compared to traditional rehabilitation methods. This protocol combines multisensory stimulation and cognitive-behavioural techniques to elicit and intensify the occurrence of adaptive responses and reduce maladaptive behavioural patterns.

Methods. A control group and an experimental group - both evaluated with the Levels of Cognitive Functioning Assessment Scale (LOCFAS) - were compared at the beginning of the rehabilitation programme and at the end of it. The control group consisting of patients assessed and treated before receiving the CBS protocol was enrolled in a traditional rehabilitation programme (only physical therapy and speech therapy). Besides the traditional therapy, the experimental group also received the CBS protocol.

Results. Patients on the CBS protocol show a greater improvement and are therefore more responsive than the control group after the 16-week remediation programme. The mean LOCFAS improvement of the experimental group is more marked during the first month of rehabilitation and is associated to the entry LOCFAS level, while in the control group the improvement on LOCFAS is considered to be 'spontaneous' and is associated to the aetiology of the brain damage.

Conclusions. Our results show a better initial outcome for patients receiving the CBS protocol.     

Conduct disorder in referred children and adolescents: clinical and therapeutic issues

OBJECTIVE: Studies on referred children and adolescents with conduct disorder (CD) have relevant implications for prevention and treatment. We addressed this issue in a large sample of youths with CD, considering age at onset, sex, and response to treatments as variables. METHODS: The sample consisted of 198 patients (153 males and 45 females; age range, 8-18 years; mean age, 13.2 +/- 2.6 years), consecutively diagnosed as having CD during a 5-year period. The diagnoses were based on fulfillment of the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria, according to historical information, prolonged observations, and a clinical interview (Schedule for Affective Disorders and Schizophrenia for School-Age Children-Present and Lifetime Version). Three subtypes of aggressive behaviors-"predatory" (controlled, planned, and goal-oriented), "affective" (impulsive, explosive, and unprofitable), and "mixed" (with both the features)-were considered in this study. RESULTS: Patients with prepubertal onset were younger at referral and had a poorer socioeconomic status. Their condition was more severe at the baseline, but their response to treatments did not differ from those with adolescent onset. Predatory and affective aggression and attention deficit hyperactivity disorder comorbidity were higher in children with prepubertal-onset CD. Regarding to sex, females were older and had a lower socioeconomic status. Their condition was more severe at the baseline and presented higher scores in self-aggression, but they responded better to treatments. Rates of attention deficit hyperactivity disorder were significantly lower in females, whereas other comorbidities (including substance abuse) were similar between sexes. Nonresponders to treatments received less frequently a psychosocial intervention, have more severe condition at the baseline, presented a more severe verbal and physical aggression, a lower affective/predatory index, and a higher rate of substance abuse. CONCLUSIONS: Age at onset and sex may be critical variables for prognosis of CD. Psychosocial intervention can significantly improve the treatment response.