Brucellosis-induced autoimmune haemolytic anaemia (AIHA).

Brucellosis is a rare zoonotic infection caused by small, fastidious Gram-negative coccobacilli of the genus Brucella that may be associated with haemolytic complications including thrombotic microangiopathy and haemolytic anaemia. We describe a patient with culture confirmed brucellosis who presented with malaise, high grade fever, hepatosplenomegaly and Coombs-positive autoimmune haemolytic anaemia. The patient was successfully treated with combination of doxycycline and rifampicin with no further episodes of relapses or haemolysis. Although rare, the possibility of brucellosis should always be kept in mind in patients with risk factors who present with haemolysis and endemic area.

Severe hypothyroidism presenting with supraventricular tachycardia.

Thyroid disease is common and can have various systemic manifestations including cardiac diseases. Hypothyroidism is commonly associated with sinus bradycardia, low QRS complexes, prolonged QT interval and conduction blocks but rarely may cause arrhythmias. We present a patient who presented with presyncope and supraventricular tachycardia with severe hypothyroidism. Patient responded well to thyroxine replacement with biochemical improvement, the disappearance of arrhythmia after restoration of euthyroidism suggests that hypothyroidism might be the cause of supraventricular tachycardia. The aim of this report is to underline the possible aetiological link between supraventricular tachycardia and hypothyroidism, although supraventricular arrhythmias are ordinary features of hyperthyroidism.

Coping with challenging behaviours of children with autism: effectiveness of brief training workshop for frontline staff in special education settings.

BACKGROUND: The present study examined the effectiveness of three staff training elements: psychoeducation (PE) on autism, introduction of functional behavioural analysis (FBA) and emotional management (EM), on the reaction of challenging behaviours for frontline staff towards children with autism in Hong Kong special education settings. METHODS: A sample of 311 frontline staff in educational settings was recruited to one of the three conditions: control, PE-FBA and PE-FBA-EM groups. A total of 175 participants completed all three sets of questionnaires during pre-training, immediate post-training and 1-month follow-up. RESULTS: Findings showed that the one-session staff training workshop increased staff knowledge of autism and perceived efficacy but decrease helping behavioural intention. CONCLUSIONS: In spite of the limited effectiveness of a one-session staff training workshop, continued staff training is still necessary for the improvement of service quality. Further exploration on how to change emotion response of staff is important.

Association of the POU class 2 homeobox 1 gene (POU2F1) with susceptibility to Type 2 diabetes in Chinese populations.

AIMS: POU class 2 homeobox 1 (POU2F1), also known as octamer-binding transcription factor-1 (OCT-1), is a ubiquitous transcription factor that plays a key role in the regulation of genes related to inflammation and cell cycles. POU2F1 is located on chromosome 1q24, a region with linkage for Type 2 diabetes in Chinese and other populations. We examined the association of POU2F1 genetic variants with Type 2 diabetes in Hong Kong Chinese using two independent cohorts. METHODS: We genotyped five haplotype-tagging single nucleotide polymorphisms at POU2F1 in 1378 clinic-based patients with Type 2 diabetes and 601 control subjects, as well as 707 members from 179 families with diabetes. RESULTS: We found significant associations of rs4657652, rs7532692, rs10918682 and rs3767434 (OR = 1.26-1.59, 0.0003 < P(unadjusted) < 0.035) with Type 2 diabetes in the clinic-based case-control cohorts. Rs3767434 was also associated with Type 2 diabetes (OR = 1.55, P(unadjusted) = 0.013) in the family-based cohort. Meta-analysis revealed similar associations. In addition, the risk G allele of rs10918682 showed increased usage of insulin treatment during a mean follow-up period of 7 years [hazard ratio = 1.50 (1.05-2.14), P = 0.025]. CONCLUSIONS: Using separate cohorts, we observed consistent results showing the contribution of multiple variants at POU2F1 to the risk of Type 2 diabetes.

Suicidal ideation among HIV+ former blood and/or plasma donors in rural China.

Suicidal ideation is life-threatening and is prevalent among people living with HIV (PLWH). A dearth of such studies was conducted in China. This study investigated the prevalence of suicidal ideation and associated factors among PLWH who were former blood and/or plasma donors (FBPD) in a rural county in central China. Prospective respondents were randomly selected from a local registry; 176 PLWH who were FBPD joined the study. With informed consent, these PLWH and their spouse were interviewed separately and anonymously. Respectively, 34 and 8% of the sampled (index) PLWH self-reported having suicidal ideation and making a suicidal attempt in the last year. The multivariate analyses results showed that the index PLWH's Physical Function subscale score of the Medical Outcomes Study HIV Health Survey scale (Odds ratio (OR)=6.67, 95% CI=1.69-26.27, 75 percentiles), the Depression subscale score of the Depression, Anxiety, and Stress Scales (DASS; OR=9.26, 95% CI=1.32-64.77), and the spouse's Depression subscale score of the DASS (OR=7.64, 95% CI=1.37-42.77) were independently associated with the index PLWH's suicidal ideation. HIV-related variables (e.g., duration of diagnosis, treatment and side effects) and perceived discrimination of the index PLWH, and HIV status of the PLWH's spouse, were not significant factors. Depression is a risk factor for suicides. Moreover, depression may be contagious and the depression status of the spouse also matters. Treatments for depression and prevention intervention for suicides targeting PLWH in rural China are not readily available. Such services are greatly warranted and need to be provided to both the PLWH and his/her spouse.

Risk and protective factors in association with mental health problems among people living with HIV who were former plasma/blood donors in rural China.

A random sample of 271 people living with HIV (PLWH) who were former plasma/blood donors and a convenience sample of 67 HIV negative villagers were anonymously interviewed. Compared with the non-PLWH, PLWH reported higher prevalence of symptoms of depression (adjusted OR = 2.53, p=0.001), anxiety (adjusted OR = 1.85, p=0.04), and stress (adjusted OR = 1.77, p=0.06). Of the PLWH respondents, 81.7% received Highly Active Antiretroviral Therapy (HAART); 32.1% of whom reported some side effects. Respectively 13.7%, 37.4%, and 38.4% PLWH perceived discrimination from their family members, relatives/friends, and neighbors. Absence of HAART, poor physical function, perceived discrimination from relatives and friends, and low level of resilience were associated with depression (stepwise regression; beta = - 0.28-0.17, R-square = 0.22), anxiety and stress (R-square = 0.32 and 0.16, respectively). The majority of respondents (70.1%) desired group intervention as a means for providing psychological support services. Relevant programs should both remove risk factors (e.g., absence of medical treatment, HIV-related discrimination) and promote protective factors (e.g., resilience). Support group is one of the potentially useful approaches to provide psychological support services.

Thyrotropin-induced formation of functional follicles in primary cultures of ovine thyroid cells.

In primary cultures of ovine thyroid cells, TSH induced the expression of several differentiated functions including the formation of follicles, and synthesis and storage of iodinated thyroglobulin in the follicular lumen. In the present report, these follicles were shown by transmission (TEM) and scanning electron microscopy (SEM) to be intact, comprised of two or more cells and to possess numerous microvilli on the inner cell membranes facing the follicular lumen. The TSH-induced formation of follicles was reversible and dynamic, with the kinetics of formation preceding that of iodination. The follicles were further demonstrated to be functional in terms of thyroglobulin storage and iodination.

Phorbol esters stimulate growth and inhibit differentiation in cultured thyroid cells.

The potent tumor promoter 12-O-tetradecanoylphorbol 13-acetate (TPA) has biological effects on cell growth and differentiation similar to the effects of epidermal growth factor (EGF) on a variety of cells. Since EGF has been shown recently to stimulate thyroid cell proliferation and inhibit iodine metabolism, we examined the effects of phorbol esters on primary ovine thyroid cultures. TPA stimulated cell growth in a manner similar to EGF. The growth effects of EGF and TPA in combination were not additive. In contrast, TPA (1.6 X 10(-7) M) was a more potent inhibitor of iodine uptake and incorporation than EGF (10(-9) M) at their maximally effective concentrations. The inhibitory effects of TPA were also more rapid and less reversible than those of EGF. TPA and EGF in combination inhibited iodine metabolism more than either agent alone at its maximally effective concentration. Both TPA and EGF reduced the accumulation of cAMP in TSH-stimulated cells, but (Bu)2cAMP and stimulators of adenylate cyclase failed to overcome TPA's inhibition of iodine metabolism. TPA interacted with EGF by reducing the affinity of membrane receptors for [125I]iodo-EGF. Although the alteration in EGF-receptor interaction induced by TPA may play a role in mediating TPA's biological effects, the additive effects of TPA and EGF on iodine metabolism suggest that TPA does not act solely through the EGF receptor-effector system. Agents other than TSH, including phorbol esters and EGF, are potent modulators of thyroid growth and differentiated function. Despite several similarities in biological activity, TPA and EGF do not modulate differentiated function in an identical manner. Both factors act at least partially through a non-cAMP-dependent pathway, providing indirect evidence of another second messenger(s) in the control of thyroid function.

Inability of anti-epidermal growth factor receptor monoclonal antibody to block "autocrine" growth stimulation in transforming growth factor-secreting melanoma cells.

Mouse monoclonal antibodies to the human epidermal growth factor (EGF) receptor were raised by immunizing with plasma membrane vesicles prepared from A431 cells. This paper describes the characterization of one of the IgG anti-receptor monoclonal antibodies generated and its use to probe the role of transforming growth factor (TGF) in the autonomous growth of a melanoma cell line in culture. This antibody blocks: 1) the binding of 125I-EGF to the A431 EGF receptor; 2) the EGF stimulation of the EGF-dependent protein kinase in vitro; and 3) human fibroblast DNA synthesis and proliferation in culture. It can precipitate the EGF receptor from metabolically labeled A431 cells and human fibroblasts and these receptors have indistinguishable peptide maps. No EGF receptor could be detected by immunoprecipitation after fibroblasts were treated with EGF or conditioned medium from the melanoma cells which secrete EGF-like TGF (alpha TGF). The antibody itself did not down-regulate the receptor but could block down-regulation caused by EGF and alpha TGF. Despite its ability to block EGF-stimulated growth and down-regulation in fibroblasts, the antibody was unable to block the growth and soft agar colony formation of alpha TGF-secreting melanoma cells, nor could the antibody detect EGF receptor in these cells under the conditions developed to prevent down-regulation and lysosomal degradation of the EGF receptor. These studies suggest that these melanoma cells do not have the intact EGF receptor and that the secretion of alpha TGF by these cells plays no role in their growth in culture. The absence of receptor cannot be explained by down-regulation by secreted alpha TGF.

Thyrotropin regulation of plasminogen activator activity in primary cultures of ovine thyroid cells.

In primary cultures of ovine thyroid cells, a high level of plasminogen activator (PA) activity was detected in the culture media. This level is much higher than in primary cultures of Sertoli cells, granulosa cells, and pituitary cells. PA activity increased with time in culture and was regulated by TSH and insulin. Activity gel analysis of the culture media revealed a major band of 43,000 daltons and a minor one of 70,000 daltons, suggesting the presence of both of the urokinase-type and the tissue-type PA in the media.

Increase of L-serine dehydratase activity under gluconeogenic conditions in adult-rat hepatocytes cultured on collagen gel/nylon mesh.

Hormonal regulation of L-serine dehydratase [L-serine hydro-lyase (deaminating), EC 4.2.1.13] was studied in primary cultures of adult-rat hepatocytes. The hepatocytes were isolated by collagenase perfusion and maintained in culture on collagen-gel/nylon-mesh substrata. L-Serine dehydratase activity was measured with [14C]threonine as substrate. The enzyme activity in hepatocytes of normal adult rats was low and declined rapidly in culture in L-15 medium containing 0.1 micro M-insulin and even more in the presence of glucose. L-Serine dehydratase activity in hepatocytes of rats with streptozotocin-induced diabetes was initially 20-fold higher than that of normal rats, but fell rapidly to a low value by 4 days in culture. Hormonal regulation of the enzyme activity was manifested by treatment of the cultured hepatocytes with insulin (0.1 micro M), glucagon (0.3 micro M), dexamethasone (10 micro M) and combinations of these hormones. Either glucagon or dexamethasone in the absence of insulin enhanced the activity of L-serine dehydratase, but failed to do so in the presence of insulin. Treatment with both hormones resulted in a 2-3-fold increase in enzyme activity in culture on days 3 and 4. Under conditions in which the enzyme activity was enhanced, glucose production by the cultured hepatocytes was concomitantly increased. Glucose production resulted in part from gluconeogenesis from pyruvate and not entirely from glycogenolysis. The gluconeogenic conditions of culture resulted in a decrease in cellular lipids in the cultured hepatocytes, as evidenced by ultrastructural studies.

Relationship between membrane fluidity and capping of receptors for concanavalin A.

The activities of a range of phenylalaninol-related compounds on capping of concanavalin A and induction of rounding of Chinese hamster ovary tsHl cells, as well as on the fluidity of phosphatidylcholine-cholesterol (1:1) liposomes, have been examined. These compounds include phenylalaninol, histidinol, leucinol, benzyl alcohol, benzylamine, 2-phenylethanol, 2-phenylamine, 3-phenyl-1-propanol, 3-phenyl-1-propylamine, and 3-phenylpropionic acid. The results indicate a strong correlation between the capacities of these compounds to enhance fluidity and their capacities to inhibit capping of concanavalin A. The specificity of this correlation is suggested by the finding that both types of capacities are poorly correlated with the capacities of the various compounds to induce cell rounding.

Accumulation of actin microfilaments in adult rat hepatocytes cultured on collagen gel/nylon mesh.

Adult rat hepatocytes cultured on a collagen gel/nylon mesh support were shown previously to exhibit fetal properties as a function of time in culture and to accumulate numerous microfilament structures as seen with electron microscopy. The present study characterizes the accumulation and properties of such microfilaments. Electron microscopy revealed that microfilaments, 40 to 60 A in diameter, accumulated as a compact network almost exclusively beneath the plasma membrane at the interface with the culture medium. Treatment with heavy meromyosin resulted in the formation of the characteristic arrowhead complexes on these microfilaments, indicating that they contain actin. Extraction of glycerinated hepatocytes with low-ionic-strength solutions removed most of the microfilaments, which could also be disrupted by treatment of the cells with cytochalasin B. The disruption was reversible, and reappearance of the network of microfilaments, though less compact, was observed as early as 30 min after removal of cytochalasin B. At later times in culture, the microfilaments developed into a more compact and extensive network, and their reappearance was not dependent on protein synthesis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins extracted from hepatocytes labeled with L-[35S]methionine or from acetone powders of the labeled hepatocytes showed a major band of a molecular weight of 42,000 that comigrated with skeletal muscle actin on the sodium dodecyl sulfate-polyacrylamide gels. The amount of radioactivity in this band increased with the age of the hepatocyte culture; this indicated an increase in the synthesis of the actin-like protein as well as a greater degree of polymerization into microfilaments. This finding was further confirmed by the demonstration of an increase in the proportion of the filamentous form of actin in the late cultures by means of a DNase I assay. The studies reported in this paper indicate that (a) the microfilaments in cultured hepatocytes contain actin, with properties similar to those found in other nonmuscle cells, and (b) the large accumulation of microfilaments is due in part to the synthesis of actin and the formation of new microfilaments.