Irma Herawati Suparto

Delineation of a CpG Phosphorothioate Oligodeoxynucleotide for Activating Primate Immune Responses In Vitro and In Vivo

Oligodeoxynucleotides (ODN) containing unmethylated CpG dinucleotides within specific sequence contexts (CpG motifs) are detected, like bacterial or viral DNA, as a danger signal by the vertebrate immune system. CpG ODN synthesized with a nuclease-resistant phosphorothioate backbone have been shown to be potent Th1-directed adjuvants in mice, but these motifs have been relatively inactive on primate leukocytes in vitro. Moreover, in vitro assays that predict in vivo adjuvant activity for primates have not been reported. In the present study we tested a panel of CpG ODN for their in vitro and in vivo immune effects in mice and identified in vitro activation of B and NK cells as excellent predictors of in vivo adjuvant activity. Therefore, we tested >250 phosphorothioate ODN for their capacity to stimulate proliferation and CD86 expression of human B cells and to induce lytic activity and CD69 expression of human NK cells. These studies revealed that the sequence, number, and spacing of individual CpG motifs contribute to the immunostimulatory activity of a CpG phosphorothioate ODN. An ODN with a TpC dinucleotide at the 5′ end followed by three 6 mer CpG motifs (5′-GTCGTT-3′) separated by TpT dinucleotides consistently showed the highest activity for human, chimpanzee, and rhesus monkey leukocytes. Chimpanzees or monkeys vaccinated once against hepatitis B with this CpG ODN adjuvant developed 15 times higher anti-hepatitis B Ab titers than those receiving vaccine alone. In conclusion, we report an optimal human CpG motif for phosphorothioate ODN that is a candidate human vaccine adjuvant.

CpG DNA overcomes hyporesponsiveness to hepatitis B vaccine in orangutans

Oligonucleotides containing immunostimulatory CpG motifs (CpG ODN) have been shown to be potent Th1-type adjuvants for augmenting antigen-specific responses in mice against hepatitis B surface antigen (HBsAg). The hepatitis B virus (HBV) infects only humans and great apes and appears to exist among wild chimpanzees and orangutans. An outbreak of HBV among orangutans being rehabilitated for re-introduction to the jungle caused the death of several animals. A prophylactic vaccination program revealed that orangutans are quite hypo-responsive to a current commercial vaccine compared to results obtained previously in humans and chimpanzees. Addition of CpG ODN to hepatitis B vaccine greatly increased the seroconversion rate and the titers of antibody against HBsAg (anti-HBs). This is the first demonstration of CpG DNA in a great ape and the results have important implications for the vaccination of humans against HBV and other diseases.

Effects of Inhaled Citronella Oil and Related Compounds on Rat Body Weight and Brown Adipose Tissue Sympathetic Nerve

Citronella oil is one of the most famous Indonesian essential oils, having a distinctive aroma. As with other essential oils, it is crucial to explore the effects of inhalation of this oil. Therefore, the aim of this research was to elucidate the effects of inhalation of citronella oil and its components isolated from Cymbopogon nardus L. (Poaceae), Indonesian local name: “Sereh Wangi” on the body weight, blood lipid profile, and liver function of rats, as well as on the sympathetic nerve activity and temperature of brown adipose tissue. Sprague-Dawley male adult rats fed with high fat diet (HFD) were made to inhale citronella oil, R-(+)-citronellal, and β-citronellol for five weeks, and the observations were compared to those of HFD rats that were not subjected to inhalation treatment. The results showed that inhalation of β-citronellol decreased feed consumption. As a consequence, the percentage of weight gain decreased compared with that in control group and the blood cholesterol level in the β-citronellol group was significantly lowered. Concentration of liver function enzymes were not significantly different among the groups. In conclusion, inhalation of citronella oil, specifically β-citronellol, decreased body weight by decreasing appetite, without any marked changes in liver enzyme concentrations.

A comparison of two progestins on myocardial ischemia-reperfusion injury in ovariectomized monkeys receiving estrogen therapy.

ObjectiveIt has been reported that the progestin medroxyprogesterone acetate (MPA), but not norethindrone acetate (NETA), inhibits the beneficial vascular effects of post-menopausal estrogen therapy, but their effects on the myocardium are unclear. The goal of this study is to compare the effects of these two progestins on post-ischemic myocardial damage. MethodsOvariectomized monkeys were fed an atherogenic diet for 18 months while receiving, or not receiving (control, n=15), the monkey equivalent to a womans dose of 5 μg ethinyl estradiol with either 1 mg NETA daily (n=15) or 2.5 mg MPA daily (n=15). The left anterior descending coronary artery was occluded for 1 h and then released to allow myocardial reperfusion for 4 h. Infarct size was quantified using the histochemical stain triphenyl-tetrazolium chloride. Regional myocardial blood flow was measured by 15 μm neutron-activated microspheres, blood pressure and heart rates with a pneumatic cuff, stroke volume by echocardiography, coronary output by thermodilution and neutrophil accumulation in the myocardium using myeloperoxidase (MPO) activity. ResultsThe infarct size (area of necrosis/area at risk) was similar between the control group (21±3%) and the MPA group (29±3%) (P<0.05) but significantly less in the NETA group (3±2%) than other groups (P<0.05). The hemodynamic myocardial function and regional myocardial blood values were similar among groups before, during and 4 h after reperfusion (all P-values >0.05). Similarly, there were no treatment effects on MPO activity (P>0.05). ConclusionsNETA, but not MPA, diminished ischemia–reperfusion injury in estrogen-treated post-menopausal females. The mechanism(s) of this difference remains unclear.

Effects of hormone therapy and dietary soy on myocardial ischemia/ reperfusion injury in ovariectomized atherosclerotic monkeys

Objective: Hormone therapy (HT) and dietary soy (Soy) inhibit myocardial ischemia/reperfusion (I/R) injury in nonatherosclerotic animals. The aim of this study was to determine their independent and interactive effects on I/R in monkeys previously fed an atherogenic diet for 15 months. Design: Ovariectomized atherosclerotic monkeys (n = 40) were divided into one of four dietary treatment groups: (1) casein as the protein source, (2) casein and added HT (the equivalent of 5 &mgr;g ethinyl estradiol + 1 mg norethindrone acetate daily), (3) Soy protein providing 141 mg total isoflavones daily, or (4) Soy + HT. After 12 months monkeys were anesthetized, and their left anterior descending coronary artery was occluded for 1 hour and reperfused for 4 hours. Infarct size was the percentage of the area at risk not staining with triphenyltetrazolium chloride. Additional measures were myocardial blood flow, stroke volume, coronary output, myeloperoxidase, and malondialdehyde. Results: There was an interactive negative effect of HT + Soy to increase infarct size from approximately 30% (in other groups) to 55% (P = 0.0004). Additionally, there were negative main effects of Soy on blood flow, coronary output, and stroke volume during I/R (all P values <0.05). There were no effects of treatment on either myeloperoxidase or malondialdehyde. Conclusions: Neither HT nor Soy had beneficial effects, whereas their combination had harmful effects, on myocardial I/R injury in monkeys with preexisting atherosclerosis. The mechanism of this negative interaction remains unclear but may relate to Soys negative effects on hemodynamics.

Gender, age and differences in stem cell expression and efficacy

Submit Manuscript | http://medcraveonline.com examples of age and gender differences in the risk of chronic conditions such as heart disease, osteoporosis and urogenital dysfunction. There are also differences in the ability of different genders and ages to respond to treatments. It stands to reason there may be age and gender differences in their ability to regenerate tissues in response to cell therapy. This review will address the evidence for and against these potential differences.

Silkworm (Bombyx mori) hemolymph unable to substitute fetal bovine serum in insect cell culture

Fetal Bovine Serum (FBS) in animal cell culture media is an important source of nutrients for cell growth. However, the harvest and collection of FBS cause bioethical concerns. Efforts to reduce and preferably replace FBS with synthetic or other natural alternatives are continually being explored. Hemolymph silkworm (Bombyx mori) contains many nutrients needed for the process of metamorphosis. Therefore, there is possibility as an alternative nutritional supplement for cell culture to reduce the use of FBS. The objective of this study was to evaluate the macrocomponent of hemolymph and the possibility as medium supplement for Spodoptera fugiperda (Sf9) cell culture. Proximate analyses showed that hemolymph contains 89.76% of water, 2.52 mg/mL carbohydrate, 2.35% fat and 55.61 mg/mL protein. Further protein analysis, it consists of 15 fractions containing molecular weight of 22 – 152 kDa. The use of hemolymph as FBS substitution in Sf9 cell culture with various concentrations was unable to maintain and sup...