Genetically engineered human umbilical cord-derived mesenchymal stem cells expressing human interleukin-12 and in vitro growth inhibition against lung adenocarcinoma cells.

INTRODUCTION: Mesenchymal stromal cells (MSCs) are promising vehicles for cancer therapy due to their homing ability and potency to be genetically manipulated through either viral or non-viral methods. Interleukin-12 (IL-12) is one of the key immunomodulatory cytokines which has anti-tumour effect. However, systemic administration of the cytokine at therapeutic dosage can cause serious toxicity in the host system due to the high systemic level of interferon-γ (IFN-γ) induced. OBJECTIVES: This study aimed to investigate the in vitro growth inhibition of genetically engineered human umbilical cord-derived mesenchymal stromal cells (hUCMSC) expressing IL-12 on H1975 human lung adenocarcinoma cells. MATERIALS AND METHODS: Both adenoviral method and electroporation which used to generate hUCMSC-IL12 were compared. The method with better outcome was selected to generate hUCMSC-IL12 for the co-culture experiment with H1975 or MRC-5 cells. Characterisation of hUCMSC and hUCMSC-IL12 was performed. RESULTS: Adenoviral method showed superior results in transfection efficiency (63.6%), post-transfection cell viability (82.6%) and hIL-12 protein expression (1.2 x 107 pg/ml) and thus was selected for the downstream experiments. Subsequently, hUCMSC-IL12 showed significant inhibition effect on H1975 cells after 5 days of co-culture. No significant difference was observed for all other co-culture groups, indicating that the inhibition effect was because of hIL-12. Lastly, the integrity of hUCMSC-IL12 remained unaffected by the transduction through examination of their surface markers and differentiation properties. CONCLUSION: This study provided proof of concept that hUCMSC can be genetically engineered to express hIL-12 which exerts direct growth inhibition effect on human lung adenocarcinoma cells.

Myosin IIA and formin dependent mechanosensitivity of filopodia adhesion.

Filopodia, dynamic membrane protrusions driven by polymerization of an actin filament core, can adhere to the extracellular matrix and experience both external and cell-generated pulling forces. The role of such forces in filopodia adhesion is however insufficiently understood. Here, we study filopodia induced by overexpression of myosin X, typical for cancer cells. The lifetime of such filopodia positively correlates with the presence of myosin IIA filaments at the filopodia bases. Application of pulling forces to the filopodia tips through attached fibronectin-coated laser-trapped beads results in sustained growth of the filopodia. Pharmacological inhibition or knockdown of myosin IIA abolishes the filopodia adhesion to the beads. Formin inhibitor SMIFH2, which causes detachment of actin filaments from formin molecules, produces similar effect. Thus, centripetal force generated by myosin IIA filaments at the base of filopodium and transmitted to the tip through actin core in a formin-dependent fashion is required for filopodia adhesion.

Assessing and Treating Cardiovascular Risk: Traditional Moderation is the Way to Go.

No abstract available.

Cardiovascular effects of common analgesics.

The clycoxygenase (COX) enzyme forms locally active prostaglandins responsible for producing inflammation and pain. Classical non-steroidal anti-inflammatory drugs (NSAID) inhibit the COX-2 enzyme that produces inflammatory prostaglandins as well as the COX-1 enzyme that produces gastric mucosa protecting prostaglandins. By specifically inhibiting only the COX-2 enzyme, coxibs thus reduce pain but do not damage the gastric mucosa. However, COX-2 at the vascular endothelium produces antithrombotic prostaglandins, and so by inhibiting COX-2 enzyme, the coxibs promote thrombosis. Rofecoxib and valdecoxib have been withdrawn because of the adverse cardiovascular events they induce. Amongst presently available coxibs cardiovascular risk is highest with enterocoxib and lowest with celecoxib. NSAIDS also increase cardiovascular events, the risk is highest with diclofenac and lowest with naproxen. Paracetamol and corticosteroids induce hypertension, while steroids also adversely affect the heart from metabolic change as well as fluid retention. Aspirin is an anti-thrombotic agent because of its ability to inhibit the COX-1 enzyme that produces the pro-aggregatory thromboxane. However, it increases gastrointestinal bleeding, can promote fluid retention and is nephrotoxic, all of which may lead to adverse cardiovascular outcomes. Patients at especially high risk of cardiovascular events from analgesic use include the elderly, and those with heart failure, hypertension, rheumatoid arthritis, chronic renal disease, chronic obstructive airway disease and previous myocardial infarction, cerebrovascular disease or peripheral vascular disease. Adverse cardiovascular events can occur within a week of initiation of analgesic treatment.

Beta-blockers for heart failure: an evidence based review answering practical therapeutic questions.

Beta-blockers are underutilised in heart failure because clinicians may be unsure whether all beta-blockers are useful, how therapy should be initiated and whether beta-blockers are contraindicated in some patients. Bisoprolol, carvedilol and metoprolol succinate have been clearly proven to reduce mortality and hospitalisation in patients with Class II to IV heart failure; limited evidence also support short-acting metoprolol tartrate and nebivolol. Initiating dose should be very low (1.25 mg bisoprolol, 3.125 mg carvedilol, 12.5 mg metoprolol succinate) and increased gradually over weeks. Treatment benefit appears proportional to magnitude of heart rate reduction and thus target dose should be the maximum tolerated for adequate bradycardia. Even in decompensated heart failure or those with coexisting bronchospasm, beta-blockers are not contraindicated although the dose may have to be reduced or withheld temporarily. The consistent trial data should reassure clinicians and encourage them to confidently initiate beta blockers in patients with systolic heart failure.

The tumor suppressor function of LECT2 in human hepatocellular carcinoma makes it a potential therapeutic target.

Vascular invasion is one of the clinicopathologic features that are associated with early recurrence of human hepatocellular carcinoma (HCC). In this study, we have employed high-density Affymetrix oligonucleotide GeneChips (Affymetrix, Santa Clara, CA) to compare the expression profiles of HCC with and without vascular invasion. Data mining of the gene expression database established revealed that leukocyte cell-derived chemotaxin-2 (LECT2) transcripts were downregulated in HCC patients with vascular invasion. Expression of LECT2 in human HCC biopsies was significantly reduced (P < 0.0001, fold change = -7.2) when compared with non-tumorous adjacent liver tissues. The reduction of LECT2 expression was significantly correlated with the early recurrent and poor prognosis of the patient (P = 0.024). To validate the ability of LECT2 to repress the growth of HCC, an adenoviral vector encoding the secreted human LECT2 (AdLECT2) was introduced into the human HCC cell lines Hep3B and PLC/PRF/5, which do not express endogenous LECT2. Over-expression of LECT2 resulted in the significant inhibition of in vitro migration and invasion of the AdLECT2-transfected HCC cells. Additionally, over-expression of AdLECT2 in subcutaneous Hep3B tumor xenografts in athymic nude mice resulted in significant inhibition of tumor growth (P < 0.05). In summary, our data not only demonstrated that LECT2 is a candidate prognostic marker of human HCC, but also that therapeutic strategies targeting LECT2 expression is a promising therapy for human HCC.

Headache diagnosis, disability and co-morbidities in a multi-ethnic, heterogeneous paediatric Asian population.

This was a questionnaire survey on headache and migraine prevalence in 2873 Singaporean schoolchildren aged 6 to 16 years. ICHD-II headache classification, disability assessment with PedMIDAS and screening of psychosocial co-morbidities with the Paediatric Symptom Checklist were conducted. Lifetime headache prevalence was high at 80.6%, migraine prevalence was 8.6% and tension headache prevalence was 10.0%. Headache and migraine prevalence was high compared with that found in other Asian studies. Factors significantly associated with headache included adolescent age (OR = 1.5 [95% CI 1.3-1.9], p < .001), female gender at primary (OR = 1.4 [95% CI 1.1-1.8], p = .003) and secondary (OR = 1.8 [95% CI 1.3-2.5], p < .001) levels and Malay ethnicity at the primary level (OR = 2.8 [95% CI 1.6-4.9], p < .001). The average PedMIDAS score for headache disability was 3.2 +/- 8.4, and migraine disability (PedMIDAS 8.1 +/- 11.2-15.2 +/- 29.6) was lower than in some studies. Self-medication (20.5%) and use of alternative therapy (59.0%) were high among chronic daily headache sufferers. Routine sleep and stress screening is recommended for children with headaches. Recognition of the influence of genetics, lifestyle and cultural factors on headache management should be emphasized.

Hypertension in a residential home for the elderly in Penang, Malaysia.

A study of residents at the Silver Jubilee Home for the Aged was conducted to determine the prevalence, awareness and control of hypertension in this elderly community in Penang, Malaysia. Prevalence of hypertension was 36%, with 81% of patients being initially aware of this diagnosis. This relatively low hypertension prevalence rate may be because residents have a fairly sheltered lifestyle with less social stress and a daily routine that incorporates adequate exercise. Similarly, the high hypertension awareness rate compared to reported figures in the community may be because residents are more regularly monitored by the attending medical care-givers. At the beginning of the study, only 34% of hypertensive patients were well controlled with a blood pressure less than 140/90 mm Hg. This proportion rose to 53% at the end of study period. Compliance is better at a residential home because medication is served by their care-givers and cost is absorbed in this charitable organization. Our study suggests that hypertension awareness and control can be reasonable for the elderly in a residential home.

Cardiovascular outcomes in the comparative hypertension drug trials: more consensus than controversy.

The comparative anti-hypertensive drug trials conducted to assess their cardiovascular protective efficacy actually produce compatible, not conflicting, results. In the last decade, there were 13 major comparative hypertension drug trials with the cardiovascular primary outcome being statistically equivalent in 11 of these 13 trials, involving over 90 percent of the randomised 168,593 patients. Where secondary outcomes favour a drug in these trials, that arm has a significantly lower treated blood pressure as in LIFE, VALUE, ASCOT and ALLHAT. Controversy occurs in seeking to attribute the benefit to drug effect; if the benefit is attributed to the lower achieved blood pressure, then the trials become consistent. The safety and value of diuretics, beta-blockers, calcium-blockers, angiotensin-converting enzyme inhibitors and angiotensin-receptor blockers in reducing blood pressure, and in reducing clinical cardiovascular outcomes, is now clearly established. Overall, the importance of tight blood pressure control in reducing cardiovascular outcomes must be emphasised. Physicians should concentrate on achieving good blood pressure control, which often requires a combination of several antihypertensive drugs.

Unusually mild phenotype of AADC deficiency in 2 siblings.

Aromatic L-amino acid decarboxylase deficiency is a rare neurotransmitter defect leading to serotonin, dopamine and norepinephrine deficiency. Affected individuals usually present in infancy with severe developmental delay, oculogyric crises and extrapyramidal movements. We present the clinical, molecular and biochemical features of a pair of siblings who presented with fatigability, hypersomnolence and dystonia and who showed excellent response to treatment. Analysis of CSF biogenic amines, plasma AADC levels and direct sequencing of the DDC gene was performed. CSF catecholamine metabolites were reduced, with elevation of 3-O-methyldopa. Plasma AADC activity was undetectable in both siblings, and decreased in their carrier parents. One missense mutation (853C>T) was found in exon 8, and a donor splice site mutation was found in the intron after exon 6 (IVS6+4A>T). Both siblings showed excellent response to MAO inhibitor and dopamine agonist treatment. This report expands the clinical spectrum of AADC deficiency and contributes to the knowledge of the genotype and phenotype correlation for the DDC gene. It is important to recognize the milder phenotypes of the disease as these patients might respond well to therapy.

Evaluation of T cells as carriers for systemic measles virotherapy in the presence of antiviral antibodies.

Neutralizing antiviral antibodies (Abs) can hinder systemic virotherapy. Here, we used activated T cells as carriers to deliver oncolytic measles viruses (MV) to multiple myeloma xenografts in the presence of anti-MV antibodies (Abs). Virus-infected T cells expressing measles H/F fusogenic envelope glycoproteins could efficiently transfer MV infection by heterofusion, even after exposure to virus-inactivating anti-MV antisera. Severe-combined immunodeficiency (SCID) mice bearing subcutaneous or disseminated human myeloma xenografts were given MV-luciferase (MV-Luc) or MV-Luc-infected T cells intravenously. Indium111 labeling indicated that 1-2% of the virus-infected T cells trafficked to tumors. Preinfected T cells fused with tumor cells in vivo and transferred MV-Luc to tumor xenografts where intratumoral viral spread was monitored non-invasively using bioluminescent imaging. In mice passively immunized with high titers of measles immune serum, intravenous virus and cell delivery were both inhibited. Decreasing the amount of measles immune serum given to mice permitted tumor infection by virus-infected T cells and cell-free virus. In conclusion, virus-loaded T cells may facilitate systemic measles virotherapy in the presence of antiviral Abs and they warrant further investigation as potential MV cell carriers.

Choice of antihypertensive drug in the diabetic patient.

The hypertensive patient with type 2 diabetes is especially at risk of adverse cardiovascular events. The United Kingdom Prospective Diabetes Study (UKPDS) and Hypertension Optimal Treatment (HOT) studies suggested that treatment to a lower target blood pressure resulted in better prevention of clinical disease in these patients. Most trials comparing antihypertensive drugs have shown only minimal differences between the various agents. The evidence from the trials suggests that diuretics, beta-blockers, calcium channel blockers (CCBs), angiotensin-converting enzyme (ACE) inhibitors, and the angiotensin-receptor antagonists (ARBs) will all successfully reduce adverse clinical events. The largest of the comparative hypertensive drug trials, the Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT), demonstrated that a diuretic has a better hypotensive effect, and was more successful in preventing many aspects of cardiovascular disease compared with CCBs and ACE inhibitors. The importance of good blood pressure control and the general equivalence of antihypertensive drugs were again shown in the Valsartan Antihypertensive Long-term Use Evaluation (VALUE) trial, which compared an ARB with a CCB. Choice of antihypertensive agent should be individualized and guided by the presence of concomitant clinical disease and the need to protect any specific target organ system in the diabetic hypertensive. Diuretics, being potent hypotensive drugs with clearly demonstrated clinical benefit, should form part of the antihypertensive regimen of most diabetic hypertensives. ACE inhibitors and ARBs are especially useful in preventing nephropathy. Most patients will require a combination of antihypertensive drugs to achieve tight blood pressure control of under 130/80 mm Hg in the diabetic hypertensive. The clinician should concentrate on seeking this lower target blood pressure rather than be excessively concerned about which is the best antihypertensive agent.

A doctor's duty is to heal the unhealthy: the story of Tun Dr Mahathir Mohamad.

Mahathir Mohamad was born in 1925 in Alor Star, Kedah. He entered the King Edward VII College of Medicine in Singapore in 1947 and graduated in 1953. His years in the medical school equipped young Mahathir with the training necessary to assess and diagnose a problem, before dispensing the appropriate treatment. Throughout his later years in the political limelight, Dr Mahathir recognised the very important role the medical college had in laying the strong foundation for his successful career. He joined UMNO in 1945, already interested in politics at the tender age of 20; he was first elected into Parliament in 1964. The vigorous expression of his candid views did not go down well during the troubled days following the 13 May 1969 racial riots and he was expelled from UMNO, his writings were banned, and he was considered a racial extremist. Nevertheless, his intellectual and political influence could not be ignored for long; he returned to Parliament in 1974, and became the fourth, and longest serving, Prime Minister of Malaysia in 1981. Dr Mahathir has found fame as a Malay statesman, and an important Asian leader of the twentieth century with much written, locally and internationally, debating his policies. This article, using Dr Mahathir's own writings, starts with his description of his early life, proceeds to look at his medical career, then touches on his diagnosis of the problems plaguing the Malays, before concluding with his views on the need to stand up to the prejudices and pressures of the Western world. Throughout his life, Dr Mahathir behaved as the ever-diligent medical doctor, constantly studying the symptoms to diagnose the cause of the ills in his community and country, before proceeding to prescribe the correct treatment to restore good health. It is a measure of his integrity and intellectual capability that he did not seek to hide his failures, or cite unfinished work in an attempt to cling to political power.