Utilization of Site-Specific Recombination in Biopharmaceutical Production

Mammalian expression systems, due to their capacity in post-translational modification, are preferred systems for biopharmaceutical protein production. Several recombinant protein systems have been introduced to the market, most of which are under clinical development. In spite of significant improvements such as cell line engineering, introducing novel expression methods, gene silencing and process development, expression level is unpredictable and unstable because of the random location of integration in the genome. Site-specific recombination techniques are capable of producing stable and high producer clonal cells; therefore, they are gaining more importance in the biopharmaceutical production. Site-specific recombination methods increase the recombinant protein production by specifically inserting a vector at a locus with specific expression trait. The present review focused on the latest developments in site-specific recombination techniques, their specific features and comparisons.     

The effect of magnesium sulfate on bleeding time and nitric oxide production in preeclamsia

Preeclampsia is a disease regarding with altered vascular reactivity leading to hypertension of the mother and metabolic alterations in the fetus. This study aimed to assess nitric oxide and bleeding time following administration of magnesium sulfate to preeclamtic patients compared to normotensive pregnant women. A total of 112 subjects (56 preeclamtic patients and 56 normotensive pregnant controls) were enrolled in this case-control study. Cases and controls were matched for age, BMI, gestational age, parity and gravidity. Total concentration of nitrite and nitrate (NOx) was measured before and during magnesium sulfate (MgSO4) treatment using a modified Griess-based method. RESULTS: Systolic and diastolic blood pressures were significantly decreased during MgSO4 treatment in preeclamtic patients (p < 0.0001). NOx levels were significantly increased in preeclamtic women after MgSO4 administration (33.7 +/- 18.5 vs. 50.2 +/- 21.6, p < 0.0001) but it was not seen in normotensive parturients (52.4 +/- 28.9 vs. 57.3 +/- 21.7, p = 0.362). The bleeding time was scarcely increased following magnesium sulfate treatment in preeclamptic patients compared to normotensive pregnant women but it was not significant (p = 0.18). In addition, there was only a significantly reverse correlation between NOx levels and systolic or diastolic blood pressure in preeclamtic parturients after MgSO4 treatment (r = -0384; p = 0.003 and r = -0.29; p = 0.03, respectively). This study demonstrates that administrating MgSO4 to preeclamtic patients induced significant changes in NOx production which had a major role in modulating vasculature changes in preeclamsia.     

The effect of waterlogging on electrochemical properties and soluble nutrients in paddy soils

This study was conducted to investigate the influence of waterlogging on availability of nutrients in paddy soils. The five soils were incubated under a waterlogged condition at 30 °C for 12 weeks. The EC, Eh, pH, NH₄, K, Na, Ca, Mg, Cl, P, Fe, and Mn of soil solutions were monitored over the waterlogged period. The Eh values generally dropped to the lowest point within 14 days of waterlogging, then increased, and reached equilibrium after 8 weeks of waterlogging. The soil pH decreased in the first 2–4 weeks of waterlogging. The EC values increased partly due to dissolution of soluble salts in the first 2 weeks. The concentrations of soluble NH₄ were significantly increased with waterlogging, reached maximum values at week 4–6, and then declined to the initial level. Waterlogging increased the concentration of soluble K, Ca, Mg, Fe, and Mn ions, the magnitudes of changes were greatly affected by soil properties. Increases in soluble Na, K, Ca, and Mg were attributed to the increase in solubility of insoluble salts and increase in competition for the exchange sites. Increases in soluble Fe and Mn induced by waterlogging were attributed to the dissolution of Fe and Mn oxides under reduced conditions.     

Effects of Dietary Ascorbic Acid on Growth Performance,Body Composition,and Some Immunological Parameters of Caspian Brown Trout,Salmo trutta caspius

A 9‐wk feeding experiment was conducted to investigate the effect of dietary ascorbic acid (AA) on growth performance and tissue compositions of Caspian brown trout. The fish (9.64 ± 0.62 g) were held in tanks at a density of 40 fish per tank. Five experimental diets were prepared by providing of 9.8, 43.8, 89.4, 188.5, and 384.2 mg AA equivalent/kg diet, respectively. At the end of the trial, an increasing trend was found on growth performance with the increase of AA supplementation. Fish fed diet supplemented with more than 188.5 mg/kg AA also had significantly higher bone collagen than fish fed diets containing 9.8 mg/kg AA (P < 0.05). According to the broken line, the dietary AA requirement of Caspian brown trout fingerlings was 177.24 mg AA/kg diet based on the weight gain, 186.97 mg AA/kg diet based on the protein tissue, and 163.95 mg AA/kg diet based on the bone collagen. Serum lysozyme and alternative complement pathway activities were also influenced by graded level of AA supplementation to the diet. The present research clearly indicated that AA is required for normal growth and enhancement of immune response in Caspian brown trout fingerlings.     

A Novel Mutation in the Aprataxin (APTX) Gene in an Iranian Individual Suffering Early-Onset Ataxia with Oculomotor Apraxia Type 1(AOA1) Disease

Background

Ataxia with oculomotor apraxia type 1 (AOA1) shows early onset with autosomal recessive inheritance and is caused by a mutation in the aprataxin (APTX) gene encoding for the APTX protein.

Methods

In this study, a 7-year-old girl born of a first-cousin consanguineous marriage was described with early-onset progressive ataxia and AOA, with increased cholesterol concentration and decreased albumin concentration in serum. PCR and direct DNA sequencing was performed after DNA extraction.

Results

Sequencing analysis revealed a novel homozygous deletion in c.643 and A>T single nucleotide polymorphism in c.641 in exon 6 of the APTX gene [ENST00000379825].

Conclusion

It seems that this region of exon 6 is probably a hot spot; however, no deletions have been reported in exon 6 yet. Key Words: Ataxia oculomotor apraxia 1 (AOA1), aprataxin (APTX), Iranian     

Barley growth,yield, antioxidant enzymes,and ion accumulation affected by PGRs under salinity stress conditions

Application of plant growth regulator (PGR) may alleviate some negative effects of environmental stresses such as salinity. A controlled environment experiment was conducted to study barley (Hordeum vulgare L. cv. Reyhane) growth, yield, antioxidant enzymes and ions accumulation affected by PGRs under salinity stress conditions at Shiraz University during 2012. The treatments were PGRs at four levels—water (as control), cycocel (CCC, 19 mM), salicylic acid (SA, 1 mM), and jasmonic acid (JA, 0.5 mM)—and four salinity levels—no stress (0.67 dS m^?1, as control), 5, 10, and 15 dS m^?1, which were arranged in a factorial experiment based on completely randomized design with four replicates. The results showed that salinity stress significantly decreased plant height, peduncle length, leaf area, ear length, grain number, dry weight, grain yield, harvest index, potassium (K⁺) accumulation, and potassium/sodium (K⁺/Na⁺) concentration ratio, which were closely associated with stress severity. However, PGRs compensated some of these negative effects, so that SA foliar application had the most ameliorative effect. Salt stress also increased Na⁺ accumulation as well as the activity of peroxidase, catalase, and superoxide dismutase (SOD). Since ion discrimination and enhanced antioxidant enzymes are associated with salt tolerance, in this experiment PGRs application might have enhanced K⁺ accumulation and antioxidant enzyme activity. The activity of SOD and K⁺/Na⁺ ratio were found to be useful in salt tolerance manipulation in barley plants.     

Factors affecting shoot and root apical meristem tissue culture of Thai supersweet corn (Zea mays var. rugosa)

During last recent years, in vitro propagation technic is widely used to produce plants with desirable traits. This experiment was conducted to produce an ideal protocol for in vitro propagation of Thai supersweet corn by using shoot apical meristem (SAM) and root apical meristem (RAM) as explants. Four-day-old germinating seedlings were used as the experimental materials on culture media supplemented with a range of auxin, kinetin, and carbohydrates. The primary establishment for SAM showed the highest percentage of survival (80%) while RAM showed the highest survival (67%) and in Murashige and Skoog (MS) media supplemented. Upon acclimatization, regenerated plantlets from shoot showed the highest survival rate (12%) with the production of 21 plantlets; however, the survival rate of plantlets from root was only 20% with the production of 9 plantlets. The efficient and economic protocol that is produced in this study can be applied as an alternative to conventional propagation method for the large-scale production of Thai supersweet corn throughout the year.     

Gaucher disease: A 10 year old girl with anemia and huge spelenomegaly (a case report)

Gaucher's disease is a rare lipid storage disorder, affecting one in 40,000-200,000 people and results from a genetic deficiency of the enzyme glucocerebrosidase (glucosylceramidase). We report a 10-year old Iranian girl with chief complaint of anemia from 8 years ago, managed for iron deficiency anemia. The patient had hepatomegaly associated with huge splenomegaly which was confirmed by sonography. No skeletal disorder was found. Bone marrow aspiration revealed typical Gaucher cells. Low level of beta-glucocerebrosidase enzyme activity confirmed the Gaucher disease. The patient is now under treatment with CEREZIME, a recombinant DNA modified form of glucocerebrosidase with good condition.     

Effects of niacin on milk production and blood parameters in early lactation of dairy cows

To investigate the effects of niacin supplementation in the diet of high producing cows at early lactation, 21 holstein dairy cows were used in this experiment. Animal were assigned in to three groups based on their milk yield and calving date soon after parturition. They were received a basal diet and 0 (group 1), 6 (group 2), 12 (group 3) g of supplementation niacin per day over a 10 weeks experimental period. Milk volume was recorded and milk samples were collected for each cow at two weeks interval for analysis of fat, protein, lactose and SNF (Solid-None Fat). Blood samples were also taken for the measurement of glucose, triglyceride, Beta-hydroxy butyrate and total protein at two weeks intervals. No significant difference were observed between milk yield, milk fat, protein, lactose and SNF content in cows received niacin compared to the control group (p > 0.05). Plasma glucose in groups 2 and 3 compared to the control were higher and this difference were statistically significant (p < 0.05). Blood triglycerides were not significantly affected by niacin supplementation. BHBA were lower in cows received niacin and this difference were significant (p < 0.05). The trend of changes in the amount of blood total protein were identical in all three groups whole the level of this factor was always higher in control group compared to the others groups. Niacin has showed an increase in the level of plasma glucose and a notable decrease in the amount of blood triglyceride, beta-hydroxy butyrate and total protein, which may be due to the effect of this vitamin on the energy metabolism in cows.     

Chemical fractionation of seven heavy metals (Cd, Cu, Fe, Mn, Ni, Pb, and Zn) in selected paddy soils of Iran

The content and fractionation of seven heavy metals (Cd, Cu, Fe, Mn, Ni, Pb, and Zn) were determined in 28 surface soil samples (0–20 cm) of agricultural topsoil from Isfahan Province in central Iran. The order of abundance of metals in the soils was Fe (1240.4 mg kg^?1) > Mn (95.7 mg kg^?1) > Pb (51.6 mg kg^?1) > Zn (23.8 mg kg^?1) > Ni (13.4 mg kg^?1) > Cu (7.0 mg kg^?1) > Cd (2.8 mg kg^?1). Iron, Mn, Ni, Pb, and Zn existed in paddy soils mainly in Fe-Mn oxides (53.6 %, 65.2 %, 40.4 %, 40.8 %, 53.3 %, respectively), whereas Cu and Cd occurred essentially as residual mineral phase (41.4 %) and carbonate (36.1 %), respectively. The mobile and bioavailable fractions of Cd, Cu, Fe, Mn, Ni, Pb, and Zn in paddy soils averaged 48.8, 20.8, 0.79, 29.2, 28.5, 41.1, and 24.8 %, respectively, which suggests that the mobility and bioavailability of the seven metals probably decline in the following order: Cd > Pb > Mn ≥ Ni > Zn > Cu ? Fe, suggesting greater contribution of anthropogenic Cd. As Cd in soil is easily accumulated by plants through the root system, the concentration of Cd in these paddy soils could be a concern to human health.