Phylogeography Approach of Diloboderus abderus (Coleoptera: Melolonthidae) in the Southern Cone of America.

Diloboderus abderus (Sturm, 1826) (Coleoptera: Melolonthidae) is a serious soil pest of corn, wheat, oat, and natural and cultivated pastures in Argentina, Paraguay, Uruguay, and southern Brazil. Despite its economic importance, the genetic diversity and population structure of D. abderus remain unknown. We sequenced a fragment of the mitochondrial gene cytochrome oxidase I region (COI), of six populations of D. abderus from the Southern Cone of America. The mtDNA marker revealed a high haplotype diversity, high pairwise FST values, and significant genetic variations among populations. No correlation was found between genetic and geographical distances, yet the most common haplotype (Dab01) was present in four out of the six populations. Analysis of molecular variance showed that most of the variation was within populations of D. abderus. Tajima's D and Fu's FS tests indicated no evidence that D. abderus populations are under recent expansion. Our results indicate that genetic-based traits will likely remain localized or spread slowly, and management strategies need to be undertaken on a small scale.

NOTCH1 mutations are associated with high CD49d expression in chronic lymphocytic leukemia: link between the NOTCH1 and the NF-κB pathways.

In chronic lymphocytic leukemia (CLL), stabilizing mutations of NOTCH1, affecting up to 10-15% of cases, have been associated to poor prognosis, disease progression and refractoriness to chemotherapy. NOTCH1 mutations are significantly overrepresented in trisomy 12 CLL, a disease subset frequently expressing CD49d, the α4 chain of the very-late-activation-4 integrin, a well-known key regulator of microenviromental interactions, and negative prognosticator in CLL. In the present study, by analysing a wide cohort of 1180 CLL, we observed a very strong association between the presence of NOTCH1 mutations and the expression of CD49d (P<0.0001), occurring also outside the trisomy 12 CLL subset. Using both the MEC-1 CLL-like cells stably transfected with the NOTCH1 intracellular domain and primary CLL cells bearing a mutated or wild-type NOTCH1 gene configuration, we provide evidence that triggering of the NOTCH1 pathway resulted in a positive CD49d expression regulation, which was driven by a NOTCH1-dependent activation of nuclear factot-κB (NF-κB). Consistently, pharmacological inhibition of the NOTCH1 and/or of the NF-κB pathways resulted in impaired NF-κB nuclear translocation with consequent down-modulation of CD49d expression. Altogether, our data link for the first time NOTCH1 mutations to CD49d expression regulation through the involvement of the NF-κB pathway in CLL.

Co-transplantation of Xenogeneic Bone Marrow-derived Mesenchymal Stem Cells Alleviates Rejection of Pancreatic Islets in Non-obese Diabetic Mice.

Bone marrow-mesenchymal stem cells (BM-MSCs) have generated a great perspective in the field of regenerative medicine, and also in the treatment of inflammatory and autoimmune diseases in the past decade due to their immunomodulatory and anti-inflammatory properties. Here, we investigated the effect of xenogeneic BM-MSCs and pancreatic islets co-transplantation obtained from Wistar rats in preventing rejection or inducing tolerance to islet transplantation in non-obese diabetic mice. Non-obese diabetic mice were treated with co-transplantation of pancreatic islets and BM-MSCs (islet + MSCs group) or pancreatic islets only (islet group). Compared to the islet group, islet + MSCs had a lower expression of inflammatory markers, such as, tumor necrosis factor- α (13.40 ± 0.57 vs. 9.90 ± 0.12, P = .01), monocyte chemoattractant protein 1 (51.30 ± 6.80 vs. 9.00 ± 1.80, P = .01), and interleukin 1ß (IL-1ß) (16.2 ± 1.65 vs. 6.80 ± 1.00, P = .04). Comparing the expression of immune tolerance markers, it is noted that animals receiving the co-transplantation showed a significantly higher expression than the islet group of IL-4 (25.60 ± 1.96 vs. 2.80 ± 0.20, P = .004), IL-10 (188.40 ± 4.60 vs. 4.55 ± 0.12, P = .0001), and forkhead box P3 (34.20 ± 1.3 vs. 1.30 ± 0.2, P = .004), respectively. These results suggest an immunomodulatory action of BM-MSC in islet xenotransplantation showing that these stem cells have the potential to mitigate the early losses of grafts, due to the regulation of the inflammatory process of transplantation.

Effects of soil tillage on Tuber magnatum development in natural truffières.

Tuber magnatum Pico, the Italian white truffle, commands the highest prices of any truffle. Despite its commercial value, it is the only edible European truffle which has not yet been successfully cultivated. Because of this, it is essential to safeguard natural truffières and to identify cultural practices to maximize their productivity. Soil tillage is successfully and extensively used in black truffle cultivation to enhance productivity, but its effects are not known on the development of T. magnatum. A recently developed real-time PCR assay was applied to evaluate the effects of tillage (10-15 cm depth) on T. magnatum mycelium in two different natural truffle grounds located in Tuscany and Emilia-Romagna. Tillage effects on bulk density, ectomycorrhizal fungal communities, and ascoma production were also assessed. Tilling significantly increased the quantity of T. magnatum mycelium which seemed to be related to an increase in soil porosity by up to 34%, and the diversity of ectomycorrhizal fungal communities. On the contrary, no significant effects were found on ascoma production. The results highlight that real-time PCR is the most reliable method for evaluating the effects of cultural practices on the development of T. magnatum in soil avoiding long-term studies on fruiting body production.

Assessment of ectomycorrhizal fungal communities in the natural habitats of Tuber magnatum (Ascomycota, Pezizales).

The ectomycorrhizal (ECM) fungal communities of four natural Tuber magnatum truffle grounds, located in different Italian regions (Abruzzo, Emilia-Romagna, Molise, and Tuscany), were studied. The main objective of this study was to characterize and compare the ECM fungal communities in the different regions and in productive (where T. magnatum ascomata were found) and nonproductive points. More than 8,000 (8,100) colonized root tips were counted in 73 soil cores, and 129 operational taxonomic units were identified using morphological and molecular methods. Although the composition of the ECM fungal communities studied varied, we were able to highlight some common characteristics. The most plentiful ECM fungal taxa belong to the Thelephoraceae and Sebacinaceae families followed by Inocybaceae and Russulaceae. Although several ectomycorrhizas belonging to Tuber genus were identified, no T. magnatum ectomycorrhizas were found. The putative ecological significance of some species is discussed.

Expression of pancreatic endocrine markers by mesenchymal stem cells from human adipose tissue.

Mesenchymal stem cells (MSCs) from human adipose tissue have a great potential for use in cell therapy due to their ease of isolation, expansion, and differentiation, besides the relative acceptance from the ethical point of view. Our intention was to isolate and promote in vitro expansion and differentiation of MSCs from human adipose tissue into cells with a pancreatic endocrine phenotype. Human adipose tissue obtained from patients undergoing abdominal dermolipectomy was digested with type I collagenase. MSCs isolated by plastic adherence and characterized by cytochemistry and FACS were expanded in vitro. MSC differentiation into an endocrine phenotype was induced over 2 to 4 months with high glucose (25 mmol/L) media containing nicotinamide, exendin-4, and 2-mercaptoethanol. Insulin and glucagon expressions were analyzed by immunofluorescence. Cells isolated from human adipose tissue and expanded in vitro expressed MSC markers as confirmed by FACS and cytochemistry. Insulin but not glucagon production by differentiated cells was demonstrated by immunofluorescence. MSCs isolated from human adipose tissue were induced to differentiate in vitro into an endocrine phenotype that expressed insulin.

Expression of pancreatic endocrine markers by mesenchymal stem cells from human umbilical cord vein.

BACKGROUND: Mesenchymal stem cells (MSCs) from human umbilical cord vein have great potential for use in cell therapy because of their ease of isolation, expansion, and differentiation, in addition to their relative acceptance from the ethical point of view. Obtaining the umbilical cord at birth does not present any risk to either mother or child. OBJECTIVE: To isolate and promote in vitro expansion and differentiation of MSCs from human umbilical cord vein into cells with a pancreatic endocrine phenotype. METHODS: Mesenchymal stem cells obtained from human umbilical cord vein via collagenase digestion were characterized at cytochemistry and fluorescent-activated cell sorting, and expanded in vitro. Differentiation of MSCs into an endocrine phenotype was induced using high-glucose (23 mmol/L) medium containing nicotinamide, exendin-4, and 2-mercaptoethanol. Expression of insulin, somatostatin, glucagon, and pancreatic and duodenal homeobox 1 was analyzed using immunofluorescence. RESULTS: Cells isolated from the umbilical cord vein were MSCs as confirmed at cytochemistry and fluorescent-activated cell sorting. Expression of somatostatin, glucagon, and pancreatic and duodenal homeobox 1 by differentiated cells was demonstrated using immunofluorescence. Insulin was not expressed. CONCLUSIONS: The MSC differentiation protocol used in the present study induced expression of some endocrine markers. Insulin was not produced by these cells, probably because of incomplete induction of differentiation.

Expression of pancreatic endocrine markers by prolactin-treated rat bone marrow mesenchymal stem cells.

BACKGROUND: Mesenchymal stem cells (MSCs) are an attractive source for generation of cells with beta-cell properties. Previous studies have demonstrated the ability of prolactin to induce an increase in beta-cell mass and maturation, which suggests beneficial effects of its use in MSC differentiation protocols. OBJECTIVE: To evaluate the expression of endocrine differentiation markers in rat MSCs treated in vitro with prolactin. METHODS: Mesenchymal stem cells from bone marrow of Wistar rats were isolated, expanded, and characterized. Differentiation of MSCs was induced in medium containing 23 mmol/L of glucose, and nicotinamide, 2-mercaptoethanol, and exendin-4, in the presence or absence of 500 ng/mL of rat recombinant prolactin. Expression of endocrine markers and prolactin receptor genes was evaluated using real-time polymerase chain reaction, and compared between culture stages and presence vs absence of prolactin in the culture medium. Expression of insulin, somatostatin, glucagon, and pancreatic and duodenal homeobox 1 was also evaluated at immunofluorescence microscopy. RESULTS: Isolated cells were mostly MSCs, as confirmed at fluorescent-activated cell sorting and cytochemistry. Pax6, Ngn-3, Isl1, NeuroD1, Nkx2.2, and Nkx6.1 exhibited varied expression during culture stages. The long form of the prolactin receptor messenger RNA was induced in prolactin-treated cultures (P < .05). The somatostatin gene was induced in early stages of differentiation (P < .05), and its expression was induced by prolactin, as confirmed using immunofluorescence. CONCLUSION: Culture of rat bone marrow MSCs in differentiation medium induces expression of pancreatic endocrine-specific genes, and somatostatin and prolactin receptor expression was also induced by prolactin.

[Validity of the Narcissism Inventory (Denecke and Hilgenstock)]. / Zur Validität des Narzissmusinventars (Denecke u. Hilgenstock).

For investigating the validity of the Narzissmusinventar 27 somatic and psychic healthy men aged between 30 and 45 were tested with the Narzissmusinventar, the Fragebogen zur Erfassung von Aggressivitätsfaktoren, the State-Trait-Angstinventar and the Fragebogen zur Messung der Kontrollambitionen. Moreover a structured interview for the evaluation of type A behavior was administered. Correlative statistical analyses showed in the sense of construct validity that the dimension of the "bedrohtes Selbst" ("threatened self") is characterized by anxiety, feelings of insufficiency and aggression. The other subscales of the Narzissmusinventar correlate only with a few variables of the other tests and the speaking behavior. These subscales evaluate the narcissistic personality traits which the other instruments we used in our investigation are not able to reflect.

A repressive coping style protecting from emotional distress in low-renin essential hypertensives.

OBJECTIVE: To investigate the relationship between the behavioural characteristics and specified subgroups of patients with essential hypertension. DESIGN AND METHODS: Fifty-four patients were classified into groups with a high (n = 9), normal (n = 35) or low plasma renin activity (n = 10), and were compared with 20 normotensive subjects by psychological tests. Standardized tests were used to measure anger expression, defensiveness and the subjects' psychological status (e.g. anxiety, depression). RESULTS: A repressive coping style, defined by a high defensiveness and low anxiety levels, was found significantly more often in patients with low than in patients with high plasma renin activity and normotensive subjects. The patients with high plasma renin activity scored significantly higher on suppressed anger, anxiety and interpersonal sensitivity than did those with low plasma renin activity. The scores of the normal plasma renin activity group were similar to those of the normotensive group. CONCLUSIONS: The results underline that there is not one hypertensive 'personality'. Whereas the patients with a high plasma renin activity appear to be more susceptible to emotional conflicts, the patients with low plasma renin activity report low emotional distress and maintain an apparently well-adjusted facade.

Ischemic ECG changes are found more often in asymptomatic men with a coronary prone behaviour pattern.

Type A behaviour has been related to coronary heart disease (CHD) as an independent risk factor. Therefore, ischemic electrocardiographic (ECG) changes may be more prominent in Type A than in Type B individuals. ECG abnormalities were assessed by the Cardiac Infarction Injury Score (CIIS), which has predictive power for sudden death. In 100 healthy men aged 30-45 yr, the CIIS was related to cardiovascular risk factors such as age, blood pressure, smoking, family history of CHD and behaviour pattern groups defined by the Structured Interview (46 Type A, 20 Type X and 34 Type B subjects). The distribution of the CIIS was different among the behaviour pattern groups (p < 0.05) and was shifted towards higher ischemic scores in Type A subjects. These findings suggest that clinically asymptomatic persons with Type A behaviour have a greater probability of suffering ischemic heart disease and possible sudden death.

Suppressed aggression accelerates early development of essential hypertension.

Predictors for early development of essential hypertension were identified in a prospective study of 98 normotensive and 23 borderline hypertensive subjects of both sexes aged 18-24 years. Baseline examination included psychological tests as well as resting and stress-induced cardiovascular and neurohormonal measurements. During the 30 (+/- 4 s.d.)-month follow up, 14 out of 98 (14%) initially normotensive subjects developed borderline hypertension, while of 23 borderline hypertensive subjects, 11 (48%) remained in the borderline group (141-159/91-94 mmHg), another five (22%) increased to greater than 160/95 mmHg and seven (30%) normalized blood pressure to less than 140/90 mmHg. In the 98 normotensive subjects, the height of casual systolic blood pressure at entry was the best predictor of subsequent borderline hypertension, correctly classifying 75% of the subjects in a stepwise discriminant analysis. Stress-induced blood pressure responses, together with measures of sympathetic nervous system activity (11%) and psychological factors (6%), were relatively weak predictors of subsequent pressure classification. In the 23 borderline hypertensives, the height of systolic blood pressure induced by mental stress was the single best predictor for sustained borderline or subsequent established hypertension, classifying 74%. When all 121 subjects were taken together, the greatest increases in blood pressure were found in those subjects who had suppressed aggression, particularly those who also had normal-high or borderline blood pressure at entry. Thus, suppressed aggression emerged as a superimposed permissive factor for a steeper trajectory or acceleration of early development of hypertension.

Psychosomatic factors in borderline hypertensive subjects and offspring of hypertensive parents.

Psychosomatic factors, sympathoneural and sympathoadrenal as well as cardiovascular mechanisms, were studied in 24 patients 18-24 years of age with borderline hypertension, 50 age-matched normotensive offspring of hypertensive parents, and 49 controls with no family history of hypertension. They were compared by projective and questionnaire-based psychological tests and their circulatory and neurohormonal reactivity to mental (Stroop color-word conflict test and arithmetic test) and physical stressors (orthostasis and bicycle ergometry test) were measured. Borderline hypertensive subjects externalized aggression less (p less than 0.05) but internalized it more (p less than 0.05) and were more submissive (p less than 0.05) when compared with controls. Offspring of hypertensive parents showed a similar but weaker pattern. Both risk groups reported more positive interactions with their parents (genetic risk subjects versus controls, p less than 0.05; borderline hypertensive patients versus controls, p = 0.08) and had higher state-anxiety levels (p less than 0.05). There were more subjective symptoms of beta-adrenergic receptor-mediated functions (e.g., tachycardia, tremor) in borderline hypertensive subjects and offspring of hypertensive parents, elevated heart rates (analysis of repeated measures, p less than 0.001), and enhanced plasma norepinephrine concentrations (p less than 0.05) when compared with controls. These findings in subjects at risk for the development of hypertension suggest that psychosomatic factors and sympathetic overactivity are involved in the early phase of hypertension.

Effects of psychological and physical covariates on plasma catecholamines in borderline hypertensives and offspring of hypertensive parents.

The interpretation of plasma catecholamine measurements may be influenced by psychological and physical factors. Therefore, catecholamine concentrations were adjusted for between-subject differences by the following possible confounding factors, i.e. body-mass index, individual maximal physical work capacity, urinary sodium excretion rates and anxiety score. Subjects were 24 borderline essential hypertensives, aged 18-24 years, 50 age-matched normotensive offspring of hypertensive parents and 49 controls with no family history of hypertension studied at rest and during mental stressors (Stroop colour-word conflict test, mental arithmetic). Borderline hypertensives had consistently higher adjusted venous noradrenaline concentrations than control subjects (p less than 0.05). Adjusted plasma adrenaline concentrations were higher in borderline hypertensive subjects than in offspring of hypertensive parents during supine rest. Despite its limitations, venous plasma noradrenaline concentrations when adjusted for work capacity, body-mass, sodium excretion and anxiety suggest enhanced sympatho-neural activity in young borderline essential hypertensives.

Suppressed aggression and hyperdynamic cardiovascular regulation in normotensive offspring of essential hypertensive parents.

To trace a possible association between suppressed aggression and enhanced responses of the cardiovascular system and plasma catecholamines in a "prehypertensive" state, two groups of 24 subjects with either normal blood pressure and no family history of hypertension or normal blood pressure and a positive family history of hypertension were matched for sex and age (18-24 years). Suppressed aggression was defined by the Rosenzweig Picture-Frustration test, a projective assessment of reactions to everyday stress. Before, during, as well as after application of mental stressors (Stroop color-word conflict test and mental arithmetic), the responses of blood pressure, heart rate, and venous plasma catecholamines were measured. Offspring of hypertensive parents with suppressed aggression (n = 10) had significantly higher heart rates and diastolic blood pressures than offspring of hypertensive parents without suppressed aggression (n = 14), using analyses of covariance for repeated measures to eliminate the influence of anxiety. Plasma norepinephrine responses tended to be greater in the offspring with suppressed aggression. The results suggest that in normotensive offspring of hypertensive parents, a behavioral pattern with suppressed aggression promotes a hyperdynamic cardiovascular state and enhances the hypertensive developmental process.

Hyperadrenergic borderline hypertension is characterized by suppressed aggression.

The effect of suppressed aggression on the reactivity of the sympathetic nervous and cardiovascular systems has been investigated in two groups of 24 subjects each with either borderline hypertension or normal blood pressure and no family history of hypertension. Groups were matched for sex and age (18-24 years). Suppressed aggression was defined by the newly standardized Rosenzweig Picture-Frustration test, a projective method assessing the reaction to everyday stress. Responses of blood pressure, heart rate, and venous plasma catecholamines were measured before and during application of mental stressors, using the Stroop color-word conflict test and mental arithmetic. In an analysis of covariance for repeated measures, which eliminates the influence of anxiety, borderline hypertensive subjects with suppressed aggression had significantly higher heart rates and diastolic blood pressures and a greater noradrenaline reactivity than borderline hypertensive subjects without suppressed aggression or normotensive subjects. Suppressed aggression may lead to a hyperadrenergic form of early borderline hypertension and thereby contribute to higher blood pressure.

Personality characteristics and renin in essential hypertension.

There may be specific personality differences between some if not all biochemically defined subgroups of patients with essential hypertension. A newly standardized projective test of reactions to frustration, a questionnaire related to such characteristics as aggressiveness and dominance, and a standardized interview were applied to 16 patients with established high renin essential hypertension and 21 patients of similar age with normal renin. High renin patients are significantly less assertive, fail to externalize their aggression, perceive frustration less and try to please others more; they have a stronger need to solve problems immediately yet tend to deny social conflicts. These tendencies are likely to create internal conflict and indicate a psychosomatic component in high renin essential hypertension.

[Aggression inhibition and increased sympathetic nervous reactivity in the development of essential hypertension]. / Aggressionshemmung und gesteigerte Sympathikusreaktivität bei der Entwicklung der essentiellen Hypertonie.

Suppressed aggression and activity of the sympathetic nervous system were assessed in two risk groups for essential hypertension, viz. 24 borderline hypertensives (18 men, 6 women, age range 18-24, 18 with a family history of hypertension) and 24 normotensive offspring of hypertensive parents, and in a control group exactly matched for age, sex and socioeconomic status. In the Rosenzweig Picture-Frustration test, both potential hypertensive groups were significantly less aggressive compared with the controls, and additionally the borderline hypertensives internalized their aggression. Plasma noradrenaline concentrations, as a marker for sympathetic nervous system activity, increased significantly before and during mental stressors (Stroop colour-word conflict test, mental arithmetic) in both groups, whereas in normotensive controls no significant increase occurred. These findings point to a psychosomatic factor in the development of essential hypertension.

Changes of immunoregulatory cells induced by psychological and physical stress: relationship to plasma catecholamines.

Lymphocyte subpopulations were measured before and after physical and psychological stress in 15 healthy subjects and correlated with plasma catecholamine and cortisol levels. During psychological stress monocytes (P less than 0.05), NK (P less than 0.01), B cells (P less than 0.05) and heart rate (P less than 0.001) increased, while catecholamines remained unchanged. With physical stress granulocytes, monocytes and all lymphocyte subsets increased significantly, although B cells rose more than T cells and T (suppressor) cells more than T (helper) cells. Thus the ratio of T/B cells and of Th/Ts cells decreased (P less than 0.001 and P less than 0.01). Adrenaline and noradrenaline concentrations increased (P less than 0.001), while cortisol remained unchanged. There was a negative relationship between adrenaline and the Th/Ts cell ratio before and after stress (P less than 0.05). Lymphocyte subpopulations from a different group of 4 healthy subjects were analysed before and after isoproterenol infusion. There was a small increase in Ts and B cells only (P less than 0.1) and a decrease of the T/B cell ratio (P less than 0.05). The predominant enrichment of circulating B, Ts and NK cells during short lasting adrenergic activation, as well as the relationship of the T cell changes to plasma adrenaline, suggest an immunoregulatory effect of the sympathetic nervous system in stress.