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Forskolin inhibits cyclin D1 expression in cultured airway smooth-muscle cells.

Accumulation of intracellular cyclic adenosine monophosphate (cAMP) has been shown to inhibit the growth of cultured airway smooth-muscle cells, but the precise mechanism underlying the antimitogenic action of cAMP in these cells is unknown. We examined the effects of forskolin, an activator of adenylate cyclase, on DNA synthesis, cyclin D1 expression, and cAMP response element-binding protein (CREB) phosphorylation and DNA binding in bovine tracheal myocytes. DNA synthesis was assessed by measurement of [3H]thymidine incorporation. Cyclin D1 protein abundance and CREB phosphorylation were assessed by immunoblotting. Cyclin D1 promoter transcriptional activation was determined by measurement of luciferase activity in cells transiently cotransfected with complementary DNAs encoding the full-length cyclin D1 promoter subcloned into a luciferase reporter and beta-galactosidase (to normalize for transfection efficiency). The binding of nuclear proteins to the cyclin D1 promoter cAMP response element (CRE) was determined by electrophoretic mobility shift assay. We found that forskolin attenuated platelet-derived growth factor-induced DNA synthesis in a concentration-dependent manner. In addition, forskolin pretreatment decreased both cyclin D1 promoter activity and protein levels. Forskolin treatment induced the phosphorylation of CREB and increased the binding of nuclear protein to the cyclin D1 promoter CRE. Finally, addition of an antibody against CREB1 induced supershift of at least one protein-DNA complex. Together, these data suggest that cAMP suppresses cyclin D1 gene expression via phosphorylation and transactivation of CREB. Further studies are needed to determine whether this is the primary mechanism of cAMP-induced growth inhibition, or whether additional pathways are also involved.

Animals↗

Frontal functions in young patients with essential tremor: a case comparison study.

Essential tremor (ET) is classified as a pure motor system disease. It has been previously reported that impairments in cognitive functions can be associated with ET. The authors assessed cognitive functions in a relatively young patient group with ET and comparison subjects. Correlations between tremor severity and regional cerebral blood flow (rCBF) and neuropsychological test performances of ET patients and comparison subjects were investigated. Sixteen patients with ET and 16 comparison subjects were assessed by a comprehensive neuropsychological test battery designed to assess global attention, language, memory, visuospatial functions, and executive functions. In 11 of 16 patients and in nine of 16 comparison subjects, rCBF was measured by technetium-99m-hexamethyl propylene amine oxime single photon emission computed tomography (technetium-99m-HMPAO SPECT). The tremor severity was quantified using the Clinical Rating Scale for Tremor (CRST). Findings revealed that ET patients differed significantly from comparison subjects on tests assessing visuospatial functions and verbal memory, whereas differences in other tests did not reach statistical significance. There was no significant difference between the rCBF of ET patients and comparison subjects. There were statistically significant inverse correlations between tremor severity and executive functions. Tremor severity was inversely correlated with bilateral frontal blood flow by technetium-99m-HMPAO SPECT. Conclusions suggest that the subclinical cognitive deficits characterized by visuospatial and verbal memory impairments and executive dysfunction may be a clinical feature of ET, and the cerebello-thalamo-frontal network may play a role in the pathophysiology of this disorder.

Adolescent↗

Cardiac hydatid cyst in a child.

Presentation of hydatid disease is uncommon in children, and cardiac hydatid cyst is rare at any age. The authors discuss a pediatric patient with cardiac hydatid cyst in the posterior wall of the left ventricle. The cyst was diagnosed by two-dimensional echocardiography and by serology. A 3 month course of mebendazole therapy was only partially effective. The patient remains asymptomatic 2 years after diagnosis. HYDATID DISEASE is endemic in the Middle East. In a study from a surgical hospital in eastern Libya, the disease accounted for 0.89 percent of all hospital admissions. The echinococcus cyst grows very slowly and unless located in a critical anatomic site, it takes many years to evolve. Consequently, presentation in pediatric patients is distinctly uncommon. In an epidemiologic study from Benghazi, only 11.7 percent of patients were under 10 years old. Cardiac hydatid disease is rare at any age, accounting for less than 2 percent of all hydatid disease. Since the introduction of two-dimensional echocardiography, more cases are being diagnosed. All the reports in the recent literature are in adult patients. We report here, however, a cardiac hydatid cyst in the left ventricular wall in a child.

Child↗

Distribution of the muscarinic K+ channel proteins Kir3.1 and Kir3.4 in the ventricle, atrium, and sinoatrial node of heart.

The functionally important effects on the heart of ACh released from vagal nerves are principally mediated by the muscarinic K+ channel. The aim of this study was to determine the abundance and cellular location of the muscarinic K+ channel subunits Kir3.1 and Kir3.4 in different regions of heart. Western blotting showed a very low abundance of Kir3.1 in rat ventricle, although Kir3.1 was undetectable in guinea pig and ferret ventricle. Although immunofluorescence on tissue sections showed no labeling of Kir3.1 in rat, guinea pig, and ferret ventricle and Kir3.4 in rat ventricle, immunofluorescence on single ventricular cells from rat showed labeling in t-tubules of both Kir3.1 and Kir3.4. Kir3.1 was abundant in the atrium of the three species, as shown by Western blotting and immunofluorescence, and Kir3.4 was abundant in the atrium of rat, as shown by immunofluorescence. Immunofluorescence showed Kir3.1 expression in SA node from the three species and Kir3.4 expression in the SA node from rat. The muscarinic K+ channel is activated by ACh via the m2 muscarinic receptor and, in atrium and SA node from ferret, Kir3.1 labeling was co-localized with m2 muscarinic receptor labeling throughout the outer cell membrane.

Animals↗

Immunocytochemical demonstration of the equilibrative nucleoside transporter rENT1 in rat sinoatrial node.

Adenosine exerts multiple receptor-mediated effects in the heart, including a negative chronotropic effect on the sinoatrial node. The aim of this study was to investigate the distribution of the equilibrative nucleoside transporter rENT1 in rat sinoatrial node and atrial muscle. Immunocytochemistry and/or immunoblotting revealed abundant expression of this protein in plasma membranes of sinoatrial node and in atrial and ventricular cells. Because rENT1-mediated transport is likely to regulate the local concentrations of adenosine in the sinoatrial node and other parts of the heart, it represents a potential pharmacological target that might be exploited to ameliorate ischemic damage during heart surgery.

Animals↗

Heterogeneous expression of Ca(2+) handling proteins in rabbit sinoatrial node.

We investigated the densities of the L-type Ca(2+) current, i(Ca,L), and various Ca(2+) handling proteins in rabbit sinoatrial (SA) node. The density of i(Ca,L), recorded with the whole-cell patch-clamp technique, varied widely in sinoatrial node cells. The density of i(Ca,L) was significantly (p<0.001) correlated with cell capacitance (measure of cell size) and the density was greater in larger cells (likely to be from the periphery of the SA node) than in smaller cells (likely to be from the center of the SA node). Immunocytochemical labeling of the L-type Ca(2+) channel, Na(+)-Ca(2+) exchanger, sarcoplasmic reticulum Ca(2+) release channel (RYR2), and sarcoplasmic reticulum Ca(2+) pump (SERCA2) also varied widely in SA node cells. In all cases there was significantly (p<0.05) denser labeling of cells from the periphery of the SA node than of cells from the center. In contrast, immunocytochemical labeling of the Na(+)-K(+) pump was similar in peripheral and central cells. We conclude that Ca(2+) handling proteins are sparse and poorly organized in the center of the SA node (normally the leading pacemaker site), whereas they are more abundant in the periphery (at the border of the SA node with the surrounding atrial muscle).

Animals↗

Expression of embryonic fibronectin isoform EIIIA parallels alpha-smooth muscle actin in maturing and diseased kidney.

In this study we examined if an association exists between expression of an alternatively spliced "embryonic" fibronectin isoform EIIIA (Fn-EIIIA) and alpha-smooth muscle actin (alpha-SMA) in the maturing and adult rat kidney and in two unrelated models of glomerular disease, passive accelerated anti-glomerular basement membrane (GBM) nephritis and Habu venom (HV)-induced proliferative glomerulonephritis, using immunohistochemistry and in situ hybridization. Fn-EIIIA and alpha-SMA proteins were abundantly expressed in mesangium and in periglomerular and peritubular interstitium of 20-day embryonic and 7-day (D-7) postnatal kidneys in regions of tubule and glomerular development. Staining was markedly reduced in these structures in maturing juvenile (D-14) kidney and was largely lost in adult kidney. Expression of Fn-EIIIA and alpha-SMA was reinitiated in the mesangium and the periglomerular and peritubular interstitium in both models and was also observed in glomerular crescents in anti-GBM nephritis. Increased expression of Fn-EIIIA mRNA by in situ hybridization corresponded to the localization of protein staining. Dual labeling experiments verified co-localization of Fn-EIIIA and alpha-SMA, showing a strong correlation of staining between location and staining intensity during kidney development, maturation, and disease. Expression of EIIIA mRNA corresponded to protein expression in developing and diseased kidneys and was lost in adult kidney. These studies show a recapitulation of the co-expression of Fn-EIIIA and alpha-SMA in anti-GBM disease and suggest a functional link for these two proteins.

Actins↗

The use of AAP-recommended disciplinary practice guidelines among African American caregivers of children in Head Start programs.

OBJECTIVE: The American Academy of Pediatrics (AAP) recommends that parents not use harsh disciplinary practices. Previous studies have characterized the disciplinary practices of African American parents as harsh, with reliance on more aggressive techniques not currently recommended by the AAP. However, recent research has indicated more disciplinary practice diversity among African Americans. This study describes factors associated with the use of AAP-recommended disciplinary practices among lower-income African American caregivers of children in Head Start. METHODS: Subjects were caregivers of children at three Head Start sites. Participants were eligible for inclusion if the biological mother, biological father, or target child was identified as African American. Using consensus methods, responses to the Parental Discipline Methods Interview (PDMI) were described as consistent or inconsistent with AAP guidelines regarding use of negative disciplinary practices (e.g., spanking, yelling). Caregivers avoiding any of these inconsistent methods were referred to as "adherent." RESULTS: "Adherent" caregivers were older (32.5 years vs. 30.4 years) and had more education (86.0% vs. 75.4% high school graduates). They were also less likely to report that their child had behavioral problems (12.9% vs. 25.2%) or deficient social skills (1.7% vs. 8.0%). CONCLUSIONS: Lower-income African American caregivers were more likely to use disciplinary practices consistent with AAP guidelines if they had higher levels of education and were living in an urban setting. Caregivers describing their child as having fewer behavior problems, better social skills, or themselves as less stressed were also more likely to be "adherent."

Adult↗

Diabetes and diet.

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Diabetes Mellitus↗

Carbamazepine and folic acid in trigeminal neuralgia patients.

The effect of carbamazepine monotherapy on the red cell folate level of 133 patients with trigeminal neuralgia was evaluated. The patient group had a significantly lower mean value of red cell folate levels compared with 110 controls. No significant correlation was found between the red cell folate levels and the mean cell volume or haemoglobin values in either the carbamazepine or control group. In addition no significant correlation was found between the red cell folate levels and drug dosage. Administration of folic acid supplements raised the mean value of red cell folate significantly. Dietary folate intake was assessed in 43 trigeminal neuralgia patients and 33 matched control patients and there was no significant difference between the groups. Patients taking carbamazepine should be advised on a well-balanced diet rich in folate as opposed to being given a routine prescription of folic acid.

Adult↗

Middle cerebral arterial thrombosis in a patient with hypofibrinogenemia, 5 days after rFVIIa and FFP infusion.

A 13-year-old female patient is presented who had hypofibrinogenemia diagnosed as von Willebrand disease at 5 years of age at another hospital. She was admitted to the department of pediatric hematology with a severe headache, vomiting, and progressive right flaccid hemiplegia and lethargy. Contrast-enhanced computed tomography scan showed subdural hematoma in posterior parietal region of the brain and impending cerebellar herniation. She was given fresh-frozen plasma (FFP) and then activated factor VII (rFVIIa), 80 microg/kg was infused for replacement of von Willebrand factor. The subdural hematoma was emergently drained. The results of coagulation tests before infusion of FFP and rFVIIa revealed hypofibrinogenemia, and FFP was given every 48 hours. The patient recovered dramatically in a few days. Five days after rFVIIa infusion, a magnetic resonance angiography-proven right middle cerebral arterial thrombosis developed. It is an interesting point of discussion whether the middle cerebral arterial thrombosis was provoked as a consequence of rFVIIa and FFP infusion.

Adolescent↗

Implementation of health education, based on ethnographic study, to increase the colostrum and decrease early solid food feeding.

Traditionally, mothers provide banana to their neonates as well as discharge their colostrum prior to breastfeeding, increasing the risk of perinatal morbidity and mortality. Health education modules, based on ethnographic study, to discourage these detrimental practices were developed for use by community leaders. Two thousand six hundred and seventy neonates were followed from birth to 28 days. Overall, neonatal banana feeding decreased 18.3% and colostrum feeding increased 32.8%. Religious leaders, who had strong community influence, had the highest health module execution. Moreover, their influence on the community was important. Overall, the use of ethnographic study data to identify specific cues to action of individuals in a community, such as community religious leaders, is an effective and appropriate method for reducing the detrimental customs of both early solid-food feeding of banana and colostrum discharge prior to breast-feeding, positively affecting community perinatal morbidity and mortality.

Adolescent↗

Choroidal effusion and hypotony caused by severe anterior lens capsule contraction following cataract surgery.

PURPOSE: The case report describes a case of severe anterior capsular contraction associated with choroidal effusion. CASE REPORT: An 81 year old female with primary open angle glaucoma underwent routine phacoemulsification cataract surgery. Eight weeks following surgery the anterior capsule opening had reduced to 3 mm in size. Intraocular pressure was found to be 4 mmHg and B scan ultrasound revealed a large choroidal effusion. Anterior capsulotomy with Nd:YAG laser was performed. At review, two weeks later, the choroidal effusion had resolved and visual acuity had recovered. DISCUSSION: The Nd:YAG laser radial relaxing capsulotomies helped relieve the capsular contraction and associated traction on the ciliary body.

Aged↗

Disposal of children's faeces and implications for the control of childhood diarrhoea.

Unhygienic disposal of children's faeces is one of the key factors responsible for the high incidence of childhood diarrhoea in developing countries. The aim of this project was to study household behaviours relating to the disposal of faeces of children under the age of five. The study was conducted in Ife South Local Government Area of Osun State, Nigeria, where 320 households, containing 540 mothers with children under the age of five were randomly selected. Data was collected by observation using a schedule specifically designed for the study. Our findings show that while over 80% of children under the age of seven months defecated on a napkin, the majority of older children (over seven months of age) defecated on either the floor of the household and their surroundings or outside in the household's vicinity. Mothers usually cleaned up the children's faeces which were then thrown into a pit latrine. In some cases dogs were used to clean up the faecal matter on the floor. In either case faecal residue remained on the floor. The authors recommend educational interventions to change the unhygienic practices relating to clean up and disposal of children's faeces. The interventions include encouraging children to defecate in plastic chamber pots, promoting the practice of hand washing with soap and water after cleaning up the children's faeces and the construction of affordable and ventilated latrines by members of the communities.

Adult↗

Bananas unite!

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Group Processes↗