Codon optimization improves heterologous expression of a <Emphasis Type="Italic">Schistosoma mansoni</Emphasis> cDNA in HEK293 cells

Differences in codon usage can seriously hamper the expression of cloned cDNAs in heterologous systems. In this study, we show that the expression of a cloned Schistosoma mansoni cDNA in cultured HEK293 cells was dramatically increased by rewriting a portion of the cDNA according to human preferred codon usage, suggesting that codon optimization is a valuable strategy for improving the heterologous expression of helminth sequences. We further describe a simple modification of a recursive PCR-based method, which allows the rewriting of long stretches of DNA sequence in a single PCR reaction. This method can be used to optimize the codon usage of virtually any DNA from helminths and other parasites.     

The high rate of prevalence of CT-detected basal ganglia calcification in neuropsychiatric (CNS) brucellosis

Abstract Of 65 cases presenting with neuropsychiatric manifestations of brucellosis (CNS-brucellosis), 9(13.8%) had CT-detected basal ganglia calcification (BGC). Of these, 5 had meningitis and 4 had psychiatric manifestations as presenting features. The diagnosis of brucellosis was made by the finding of consistent history and physical findings and the presence of significantly elevated antibody litres and/or positive culture in the blood and/or CSF. In all the cases, BGC was in the form of punctate hyperdense non-enhancing shadows with average density 44.5–58.4 and maximum density 49–64HU The calcification was unilateral in 3 cases, bilateral and symmetrical in 4 and bilateral but assymmetrical in 2. None of the cases had other predisposing conditions to BGC and in none of the cases did specific anvtibrucella treatment effect a detectable change in the BGC. The finding of CT-detected BGC in patients coming from areas endemic for brucellosis should alert physicians to the possibility of underlying brucellar infection.     

Antithrombotics in thrombosis and cancer

Many cancer patients reportedly are in a hypercoaguable state, with recurrent thrombosis due to the impact of cancer cells and chemotherapy or radiotherapy on the coagulation cascade. Studies have demonstrated that unfractionated heparin (UFH) or low-molecular-weight heparin (LMWH) interfere with various processes involved in tumor growth and metastasis. These include fibrin formation, binding of heparin to angiogenic growth factors such as basic fibroblast growth factor and vascular endothelial growth factor, modulation of tissue factor and, perhaps, other more important modulatory mechanisms such as enhanced tissue factor pathway inhibitor (TFPI) release and inhibition of various matrix-degrading enzymes. Clinical trials have suggested a clinically relevant effect of LMWH, compared with UFH on the survival of cancer patients with deep vein thrombosis. Similarly, the impact of warfarin on the survival of cancer patients with thromboembolic disorders was demonstrated. Studies from our laboratory demonstrated a significant role for LMWH, warfarin, anti-VIIa and LMWH-releasable TFPI on the regulation of angiogenesis, tumor growth, and tumor metastasis. Thus, modulation of tissue factor/VIIa noncoagulant activities by LMWH, warfarin, anti-VIIa or TFPI might be a useful therapeutic option for the inhibition of angiogenesis associated with human tumor growth and metastasis.     

Long term prognosis of sarcoidosis in Arabs and Asians: predictors of good outcome

BACKGROUND: The prognosis of sarcoidosis is variable and often difficult to predict. Our aim was to identify predictors of good prognosis in Arabs and Asians with sarcoidosis. METHODS: Data on patients with sarcoidosis followed up for at least 3 years in two major hospitals in Kuwait were collected retrospectively for the period 1983 to 1995 and prospectively from 1995. RESULTS: Of the total 115 patients, 60% were females and 80% were Arabs. Majority, 86.9%, of the patients had either Stage I or II disease. Forty-five (43.7%) were followed up for 3 to 5 years, 43 (41.7%) for 5 to 9 and 15 (14.6%) for 10 or more years while 12 were lost to follow up. Good prognosis was seen in 53 (51%), intermediate in 33(32%) and poor in 17 (17%) patients. Two patients (1.9%) died. Good prognosis was observed in 74.4% of patients with Stage I, 40% of patients with Stage II and 16.7% with stage III disease, p = 0.001. In addition, presence of arthralgia predicted a good prognosis, p = 0.014. Hypercalcemia was noted only in patients with poor or intermediate outcome. Gender, ethnicity, and presence of erythema nodosum were not predictors of prognosis in our patients. Multivariate logistic regression analysis confirmed that early stage of the disease [OR (95 %CI), 6.1 (2.3-15.7), p = 0.001] and presence of arthralgia, [OR (95%CI), 4.5 (1.3-15.4), p = 0.02] were predictors of good prognosis. CONCLUSION: Presence of arthralgia and early stage of the disease were the most important predictors of good prognosis. Sex, age, ethnicity and presence of erythema nodosum did not influence the prognosis.