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To date, the success of conventional chemotherapy, radiotherapy, and targeted biological therapies in cancer treatment is not satisfactory. The main reasons for such outcomes rely on low target selectivity, primarily in chemo- and radiotherapy, ineffectiveness to metastatic disease, drug resistance, and severe side effects. Although immune checkpoint inhibitors may offer better clinical promise, success is still limited. Since cancer is a complex systemic disease, the need for new therapeutic modalities that can target or block several steps of cancer cell characteristics, modulate or repolarize immune cells, and are less toxic to healthy tissues is essential. Of these promising therapeutic modalities are pleiotropic natural products in which scorpion venom (SV) is an excellent example. SV consists of complex bioactive peptides that are disulfide-rich of different peptides' length, potent, stable, and exerts various multi-pharmacological actions. SV peptides also contain ion channel inhibitors. These ion channels are dysregulated and overexpressed in cancer cells, and play essential roles in cancer development and invasion, as well as depolarizing immune cells. Furthermore, SV has been found to induce cancer cell apoptosis, and inhibit cancer cells proliferation, invasion, metastasis, and angiogenesis. In the current review, we are presenting data that show the pleiotropic effect of SV against different types of human cancer as well as revealing one potential anticancer agent, Rhopalurus princeps venom. Furthermore, we are addressing what is needed to be done to translate these potential cancer therapeutics to the clinic.

Mona Bustami,1 Khalid Z Matalka,2 Eyad Mallah,3 Luay Abu-Qatouseh,1 Wael Abu Dayyih,3 Nour Hussein,4 Nayef Abu Safieh,4 Yousef Elyyan,5 Nagham Hussein,6 Tawfiq Arafat7 1Department of Pharmacology and Biomedical Sciences, University of Petra, Amman, 11196, Jordan; 2Faculty of Health Sciences, American University of Madaba, Madaba, Jordan; 3Department of Pharmaceutical Medicinal Chemistry and Pharmacognosy, Faculty of Pharmacy & Medical Sciences, University of Petra, Amman, 11196, Jordan; 4School of Medicine, University of Jordan, Amman, Jordan; 5Obstetrics and Gynecology, Jordan University Hospital, Amman, Jordan; 6Istishari Hospital, Amman, Jordan; 7Jordan Center for Pharmaceutical Research, Amman, JordanCorrespondence: Mona Bustami Email mbustami@uop.edu.joObjective: The study aimed to investigate the prevalence of obesity among Jordanian women and its association with a wide range of chronic diseases.Methods: Subjects were enrolled in the present cross-sectional study based on a random drop-off technique at the Obstetrics and Gynecology clinics at Jordan University Hospital. Initially, any female 18 years of age and older was asked to enroll in the study. Relevant data were gathered using a questionnaire composed of 30 questions, and body mass index (BMI) was determined from each participant’s weight and height. The following variables were collected: socio-demographic, chronic diseases, and health status. Each variable’s frequencies were reported, and the 95% confidence interval (95% CI) for each variable was calculated. For association analysis, Chi-square analysis was performed with an odds ratio (OR) and 95% CI. Multinomial logistic regression analysis was applied to a combination of independent variables and a dependent condition with covariate factors.Results: The age-standardized prevalence of overweight/obese Jordanian women was 70.6% (95% CI 66.0– 74.8%). On the other hand, the age-standardized prevalence of only obese women was 36.4 (95% Cl 31.9– 41.2%). Furthermore, the association between age and overweight/obesity was significant (p< 0.0001). The percentage of overweight and obesity started to be significant in the 30– 39 year age group. Moreover, the OR for obesity ranged from 2.7 to 7.0 (p< 0.05– 0.01) for those women with only elementary education. Besides, high parity was significantly associated with obesity and elementary education. For chronic conditions, the percentages of hypertension, diabetes, hypertriglyceridemia, osteoporosis, and rheumatoid arthritis were significantly correlated with increased BMI in Jordanian women. With age adjustment, however, only hypertension was associated with obese level 3 with OR of 7.2 and 95% CI of 2.1– 25.1 (p< 0.01).Conclusion: There is a high prevalence of overweight/obesity among women in Jordan, which was related to high parity and low education level. This high prevalence of obesity increased the incidence of chronic diseases, such as hypertension. Therefore, community-based multiple strategies are required to combat obesity in Jordanian women.Keywords: women, obesity, hypertension, chronic diseases, parity, education, Jordan

Mona Bustami,1 Khalid Z Matalka,2 Yousef Elyyan,3 Nagham Hussein,4 Nour Hussein,5 Nayef Abu Safieh,5 Fida Thekrallah,3 Eyad Mallah,6 Luay Abu-Qatouseh,1 Tawfiq Arafat7 1Department of Pharmacology and Biomedical Sciences, Faculty of Pharmacy and Medical Sciences, University of Petra, Amman 11196, Jordan; 2Matalka’s Scientific Writing, Lexington, MA, USA; 3Department of Obstetrics and Gynecology, Jordan University Hospital, Amman, Jordan; 4Istishari Hospital, Amman, Jordan; 5School of Medicine, University of Jordan, Amman, Jordan; 6Department of Pharmaceutical Medicinal Chemistry and Pharmacognosy, Faculty of Pharmacy and Medical Sciences, University of Petra, Amman 11196, Jordan; 7Jordan Center for Pharmaceutical Research, Amman, JordanCorrespondence: Mona Bustami Email mbustami@uop.edu.joObjective: The aim of this study was to assess factors related to the onset of premature/early natural menopause among Jordanian women.Methods: A cross-sectional study was conducted in early 2016. Subjects were enrolled based on random drop-off technique to the Obstetrics and Gynecology clinics at the Jordan University Hospital. Women 18 years of age and above were initially eligible to enroll, and women who had surgically induced menopause or specific disease were excluded from the analysis. Relevant data were collected using a questionnaire that included 30 questions. The following variables were collected: socio-demographic, body mass index, chronic conditions, diseases, reproductive characteristics, and health status. Hormone indicators of menopause were tested by measuring estrogen (E2) and follicle-stimulating hormone (FSH) levels. Age at natural menopause (ANM) was self-reported retrospectively and considered an independent variable against BMI, smoking, hormone therapy, and concomitant diseases. Association analysis and multinomial logistic regression were used to examine the associated factors of ANM with adjusted odds ratios (ORs), and their 95% confidence intervals (CIs) were reported.Results: A total of 409 women were included in the analysis, aged between 20− 75 years. The mean ANM in our sample was 48.5± 5.0, with 2.7% of the women experienced premature menopause (ANM < 40) and 7.8% early menopause (ANM 40– 44). Within the menopause women (n=242), the percentage of women who had premature menopause was 4.5%, 13.6% with early menopause, and 21.1% with late menopause (ANM > 52). Smoking was the major risk factor for premature/early menopausal age among Jordanian women with an OR of 2.46 (95% CI: 1.08– 5.59, p< 0.05). On the other hand, women with occasional arthritis symptoms and diseases such as hypertension, diabetes, dyslipidemia, and their combination were associated with average (45– 52 years) or late menopause (> 52 years).Conclusion: Smoking is the main contributor of premature/early menopause in Jordanian women. Increased awareness and public health policy about the adverse effects of smoking on women’s reproductive health are needed.Keywords: Jordanian women, menopause age, smoking, diseases, socioeconomic variables

During the cyclization reaction of benzyl alcohol containing amides, using NaN3 and SiCl4, additional unique chlorination development was observed to yield the novel azoles 1-[4-chloromethylphenyl]-5-[4-(methylsulfonyl)benzyl]-1H-tetrazole (3a) and 1-[4-chloromethylphenyl]-5-[4-(aminosulfonyl)phenyl]-1H-tetrazole (3b). Control experiments showed that the SiCl4 or SiCl3N3 has the major role for such functional group transformation in such a clean reaction and quantitative yield. Their molecular structures have been ascertained using the X-ray crystallography technique in addition to the spectroscopic analyses. Both compounds 3a and 3b crystallize in the monoclinic space group P21/c. The cell parameters of azole 3a are a = 22.3827 (8), A, b = 5.1602 (2) A, c = 13.4994 (5) A3, β = 95.2352 (14)°, V = 1552.67 (10) A3, and Z = 4. While the cell parameters of azole 3b are a = 20.582 (2), A, b = 5.8947 (7) A, c = 13.0796 (16) A3, β = 104.376 (4)°, V = 1537.2 (3) A3, and Z = 4. The central tetrazole ring of both compounds is planar and bears (4-chloromethylphenyl) at position one (N-1) of the central moiety. However, the substituents 4-(methylsulfonyl)phenyl and 4-(aminosulfonyl)phenyl of azoles 3a and 3b, respectively, are attached to the C-5 of the same central unit. The phenyl rings at N-1, (C2 C7) and C9 C14 in (3a); (C2 C7) and (C8 C14) in (3b) are inclined compared to the tetrazole ring with dihedral angles of 21.74° and 83.94° in 3a and 25.85° and 65.13° in 3b. The two phenyl rings, at N-1 and C-5, are rotated against each other by 87.73° (in 3a) and 72.21° (in 3b). In the crystal, intermolecular interactions between molecules of 3a,b are dominated by C H⋯O and C H⋯N hydrogen bonds. Additional Cl…π interactions add extra supramolecularity. All intermolecular interaction motifs consolidate a three dimensional network lattice. The molecular docking studies were carried out to understand the interaction of compounds 3a and 3b within the active site of the cyclooxygenase-2 enzyme, followed by a comparison study with the celecoxib drug as a reference compound. The in vitro bioassay screenings of azoles 3a and 3b showed that both compounds have poor selectivity and weak inhibition potency toward cyclooxygenase-2 enzyme.

Propranolol (PROP) undergoes extensive first-pass metabolism by the liver resulting in a relatively low bioavailability (13-23%); thus, multiple oral doses are required to achieve therapeutic effect. Since some studies have reported that glucosamine (GlcN) can increase the bioavailability of some drugs, therefore, it is aimed to study whether GlcN can change the pharmacokinetic parameters of PROP, thus modulating its bioavailability. When PROP was orally co-administered with GlcN (200mg/kg) to rats, PROP area under curve (AUC) and maximum concentration (Cmax) were significantly decreased by 43% (p

To date, the success of conventional chemotherapy, radiotherapy, and targeted biological therapies in cancer treatment is not satisfactory. The main reasons for such outcomes rely on low target selectivity, primarily in chemo- and radiotherapy, ineffectiveness to metastatic disease, drug resistance, and severe side effects. Although immune checkpoint inhibitors may offer better clinical promise, success is still limited. Since cancer is a complex systemic disease, the need for new therapeutic modalities that can target or block several steps of cancer cell characteristics, modulate or repolarize immune cells, and are less toxic to healthy tissues is essential. Of these promising therapeutic modalities are pleiotropic natural products in which scorpion venom (SV) is an excellent example. SV consists of complex bioactive peptides that are disulfide-rich of different peptides’ length, potent, stable, and exerts various multi-pharmacological actions. SV peptides also contain ion channel inhibitors. These ion channels are dysregulated and overexpressed in cancer cells, and play essential roles in cancer development and invasion, as well as depolarizing immune cells. Furthermore, SV has been found to induce cancer cell apoptosis, and inhibit cancer cells proliferation, invasion, metastasis, and angiogenesis. In the current review, we are presenting data that show the pleiotropic effect of SV against different types of human cancer as well as revealing one potential anticancer agent, Rhopalurus princeps venom. Furthermore, we are addressing what is needed to be done to translate these potential cancer therapeutics to the clinic.

1-[4-(2-Chloroethoxy)phenyl]-5-[4-(methylsulfonyl)phenyl]-1H-tetrazole (2) and 2-(4-(5-(4-(methylsulfonyl)phenyl)-1H-tetrazol-1-yl)phenoxy)ethyl nitrate (3) have been easily synthesized and their molecular structure have been determined using the X-ray crystallography technique. Both tetrazoles 2 and 3 crystallize in the monoclinic space group P21/c. The cell parameters determined for tetrazole 2 are a = 25.067 (2) A, b = 5.2236 (5) A, c = 13.5324 (13) A3, β = 102.3881 (13)°, V = 1730 (3) A3, and Z = 4. Its structure refines to R1 = 0.0654 (for 2945 observed reflections [Fo2 ≥ 2 σ(Fo2)]) and wR2 = 0.1557 (for all 4278 unique reflections). The cell dimensions for tetrazole 3 are a = 26.4012 (10) A, b = 5.14915 (18) A, c = 13.2950 (5) A3, β = 92.6313 (16)°, V = 1805.47 (12) A3, and Z = 4. Its structure refines to R1 = 0.0329 (for 3469 observed reflections [Fo2 ≥ 2 σ(Fo2)]) and wR2 = 0.0871 (for all 3657 unique reflections). For both structures, the aryl rings at the 1- and 5-positions of the tetrazole rings show no conjugation to the tetrazole group, and intermolecular interactions between adjacent molecules are dominated by weak (2.3–2.7 A) C–H⋯O and C–H⋯N contacts. The molecular docking studies were carried out to understand the interaction of compounds 2 and 3 within the active site of the cyclooxygenase-2 enzyme, followed by a comparison study with the celecoxib drug as a reference compound. The in vitro bioassay studies of tetrazoles 2 and 3 toward cyclooxygenase-1 and cyclooxygenase-2 enzymes showed that both compounds have no inhibition potency for either enzyme.

Two new proanthocyanidin trimers have been isolated from Cistus incanus herb; gallocatechin-(4α→6)-gallocatechin-(4α→8)-gallocatechin (compound 1) and epigallocatechin-3-O-gallate-(4ß→8)-epigallocatechin-3-O-gallate-(4ß→8)-gallocatechin (compound 2). The structures were determined on the basis of 1D- and 2D-NMR (HSQC, HMBC) of their peracetylated derivatives, MALDI-TOF-MS and by acid-catalysed degradation with phloroglucinol. A more abundant proanthocyanidin oligomer was also isolated, purified and its chemical constitution studied by (13)C-NMR and phloroglucinol degradation. The mean molecular weight of the polymer was estimated to be about 7 to 8 flavan-3-ol-units with a ratio of procyanidin : prodelphinidin units at 1:5, some of which are derivatised by gallic acid. Water extract and higher oligomeric proanthocyanidin fractions of C. incanus significantly inhibited TPA-induced oedema when applied topically at doses of 0.5 and 1 mg/ear in mice. Furthermore, the extracts and the pure compounds inhibited COX-1 and COX-2 activities. In addition, compound 2 exhibited an IC50 of 4.5 μM against COX-2 indicating its high selectivity towards COX-2.

Aminoacetylenic isoindoline-1,3-dione derivatives were synthesized from the reaction of potassium phthalimide with propargyl bromide to generate 2-(prop-2-yn-1-yl)isoindoline-1,3-dione (ZM1). Treatment of 2-(prop-2-yn-1-yl)isoindoline-1,3-dione with appropriate cyclic amines through Mannich reaction yielded five desired aminoacetylenic isoindoline-1,3-diones called, ZM2–ZM6. The IR, NMR and elemental analysis were consistent with the assigned structures. These synthetic compounds, except ZM6, produced significant (p ZM5 > ZM4 > ZM2. These percent inhibitions for ZM3 and ZM5 were not significantly different than those of induced by Ibuprofen, Diclofenac and Celecoxib. At 20 mg/kg dose, ZM4 produced a statistically significant reduction of inflammation (p

Plants have been used since antiquity to treat and prevent diseases. Plantain (Plantago lanceolata L.) is traditionally used for the treatment of the common cold and associated symptoms such as cough. This study was designed to evaluate the oral toxicity of plantain leaf extract-containing syrup. In preparation of the toxicological examination and to ensure the quality of the herbal preparation, analytical methods were developed and validated, and stability testing was performed. Physicochemical and microbial quality, thin layer chromatography patterns and high performance liquid chromatography fingerprints complied with the specifications during the entire period of stability testing. The marker substance, acteoside, remained within the stability-defining limits of 90%–110% for quantitative determinations. No hint of toxicity emerged from 14-day repeat dose toxicity testing in rats. The animals were given doses of 3, 6, or 12 mL of syrup per kg body weight by gavage twice daily. All animals showed normal appearance and behavior. Body and organ weights at the end of the study were similar to those in the control group. Overall, P. lanceolata syrup was found to be stable and non-toxic under the test conditions.

A novel class of modified 1,5-disubstituted tetrazoles was designed and synthesized, their biological activity as cyclooxygenases inhibitors was screened, and their molecular docking studies were performed. The structural modifications of the first category included the 4-methylsulfonyl phenyl at C-1 of the central moiety and the linkers (–OH, –CH2OH, –CH2CH2OH) with different lengths at the para position of the N-1 phenyl group. For the second category, the 4-methylsulfonyl phenyl group at C-1 was replaced with 4-aminosulfonyl phenyl. While for the third category, a methylene unit was inserted between the C-1 of the tetrazole central ring and the 4-(methylsulfonyl)phenyl group, keeping the same linkers of various extensions at the para position of the N-1 phenyl group. Among the screened compounds, tetrazole 4i showed the best inhibition potency and selectivity values for both COX-2 enzyme (IC50 = 3 μM, SI > 67) and COX-1 isoenzyme (IC50 > 200 μM). Compounds 4e, 4h, and 4i, which have the highest inhibition potency toward COX-2 were selected for the molecular docking studies to verify their inhibition and selectivity for COX-2 over COX-1 with their modified structure. The obtained theoretical studies are in agreement with the in vitro bioassay screening results, which supports the importance of the structural modifications for our studied compounds.

Eriobotrya japonica (Thunb.) Lindl. (Loquat) (EJ) has been used as a medicinal plant to treat chronic bronchitis, coughs, phlegm, high fever and gastro-enteric disorders. Since the traditional use of EJ is related to modulating inflammation processes, our earlier studies on EJ leaves were performed on the water extract to investigate specific cytokines’ modulation. These earlier studies, however, have shown that EJ leaf water extract (WE) and the water phase (WP) induce cytokines’ production in in vitro and in vivo models. Therefore, the aim of this study was to specify the group(s) of compounds in EJ leaves that have this immunomodulatory activity and their mechanism of action. WE was obtained from boiling the leaves followed by butanol extraction, yielding a butanol-water phase (WP). WP was then subjected to methanol:acetone fractionation, yielding upper (MAU) and lower (MAL) phases. For further fractionation, MAU was subjected to column chromatography followed by elution with ethanol:water (EW), methanol:ethanol (ME) and, lastly, acetone:water (AW), respectively, to reveal three sub-fractions; MAU-EW, MAU-ME and MAU-AW. MAU-AW significantly increased IFN-γ production from unstimulated and stimulated mouse spleen cells, as well as CD3+ T cells and natural killer cells. Furthermore, the fold increase of IFN-γ production by MAU-AW was concentration dependent, higher than the parent extract or any of the other sub-fractions, and such an IFN-γ increase was reversed by two JAK-STAT inhibitors. In addition, MALDI-TOF-MS analysis of the extracts and sub-fractions showed compounds with molecular weights of >500 Daltons. The MAU-AW sub-fraction contained more polar compounds, such as flavonol and caffeic glycosides. In conclusion, these polar compounds in the EJ extract are responsible for inducing IFN-γ production. Further chemical elucidation is warranted to lead to a specific IFN-γ inducer and an immunomodulator in polarizing immune cells and balancing immune responses in certain diseases.

Food or drinks may significantly alter the pharmacokinetics and pharmacodynamics of drugs which may lead to adverse events. A drug such as metformin is widely used to regulate plasma glucose level and pomegranate and licorice have been identified to help in type-2 diabetes management. However, the interactions of the latter on metformin pharmacokinetics were not studied. Therefore, we aimed here to study the impact of pomegranate and licorice on metformin pharmacokinetics in rats.Juices were given to rats for two days and half an hour before metformin (20 mg/kg) oral administration. Blood samples, then, were collected at different time intervals, processed and analyzed using validated reliable HPLC method. Plasma profile and pharmacokinetic parameters were calculated for each group.Pre-administration of pomegranate significantly reduced metformin maximum plasma concentration from 1410 to 1031 ng/ml. On the other hand, pre-administration of licorice significantly delayed metformin reaching its maximum plasma concentration. In conclusion, pre-administration of pomegranate may potentially reduce efficacy of metformin while licorice might delay metformin action. Thus, both juices should be cautiously administrated with metformin, the mainstay drug for type-2 diabetes mellitus management.

The reported pharmacological activities of acetylenic and phthalimide groups promoted our interest to synthesize a novel series of N-[4-(t-amino-yl)-but-2-yn-1-yl] isoindoline-1,3-diones as anti-inflammatory compounds. The aim of this research is to investigate the selectivity of two compounds, ZM4 and ZM5, on inhibiting cyclooxygenase (COX) in vitro and in silico as well as reducing carrageenan-induced edema in rats. Oral administration of 5-20 mg/kg ZM4 and ZM5 reduced significantly carrageenan-induced edema in dose-and time dependent manner. Furthermore, the IC50 values induced by ZM4 and ZM5 were in the range of 3.0-3.6 μM for COX1 and COX 2 but were higher than those induced by Diclofenac and Celecoxib, respectively. Docking of ZM4 and ZM5 in both COX enzymes, on the other hand, exhibited the conventional binding modes that are usually adopted by different non-steroidal anti-inflammatory drugs (NSAIDs). Furthermore, ZM4 and ZM5 bind to COX enzymes as strongly as Flurbiprofen and Celecoxib. In conclusion, aminoacetylenic isoindoline 1, 3-dione compounds have shown anti-inflammatory activity by inhibiting COX-1 and COX-2 enzymes. Interestingly, the best hits showed inhibition at low micromolar levels although they are not selective at this stage. Further research will be conducted to improve both selectivity and potency.

A new trimeric proanthocyanidin, epigallocatechin-3-O-gallat-(4β→8)-epigallocatechin- (4β→8)-catechin (1), was isolated together with three known flavan-3-ols, catechin (2), epicatechin (3), and epigallocatechin (4), and three dimeric proanthocyanidins, 5 - 7, from the air-dried leaves of Mangifera indica. Their chemical structures were determined on the basis of 1D- and 2D-NMR spectra (HSQC, HMBC) of their peracetylated derivatives, MALDITOF- mass spectra, and by acid-catalyzed degradation with phloroglucinol. The isolated compounds 1 - 7 were in vitro tested for their inhibitory activities against COX-1 and COX-2. Compound 1 was found to have a potent inhibitory effect on COX-2, while compounds 1 and 5 - 7 exhibited moderate inhibition against COX-1.

Aims of the study Eriobotrya japonica leaves had been used traditionally for the treatment of diabetes mellitus by immersing the dried leaves in a hot water drink. Few studies have shown the hypoglycemic effect of Eriobotrya japonica using crude alcoholic extract and isolated methanolic compounds. These studies proposed that the mechanism of action could be by stimulating the β-islets of Langerhans to secrete insulin, however with no scientific evidence. Methods Eriobotrya japonica water extract (EJWE) and the compounds derived from it: cinchonain Ib, procyanidin B-2, chlorogenic acid and epicatechin, were tested for their effects on insulin secretion from INS-1 cells and following oral administration in rats. Results The present study showed that EJWE increased significantly (p

The aim of this study is to investigate the involvement of an efflux pump in the development of Pseudomonas aeruginosa resistance to zinc pyrithione (ZnPT). In the presence of efflux inhibitor carbonyl cyanide m-chlorophenyl-hydrazone (CCCP), the minimum inhibitory concentration of ZnPT for P. aeruginosa resistant cells is reduced significantly (p < 0.05). In addition, the concentration of ZnPT excluded by the resistant bacteria was reduced significantly (p < 0.01). However, the above reductions did not reach the levels measured for P. aeruginosa PAO1 sensitive strain. Furthermore, such changes in P. aeruginosa resistant cells were correlated with the overexpression of outer membrane proteins, reduced sensitivity toward imipenem (p < 0.01) and increased sensitivity toward sulphatriad and chloramphenicol (p < 0.05). In a continuation to a previous study, we conclude that P. aeruginosa resistance to ZnPT is multifactorial and involves induced efflux systems.

SUMMARY Background and objectives: Most of the pharmacokinetic (PK) parameters for enalapril and enalaprilat were established following determination of the drug and its metabolite, using angiotensin converting enzyme (ACE) inhibition assays. In these methods, enalapril has to be hydrolysed to enalaprilat first and then assayed. The purpose of this study was to re-estimate the PK parameters of enalapril and enalaprilat in healthy volunteers using two specific enzyme immunoassays for enalapril and enalaprilat. Methods: The rate and extent of absorption of enalapril and enalaprilat from a 10-mg dose of two enalapril maleate commercial brands (Renetic and Enalapril) were estimated using a two-way-cross over design with 1-week washout period. Blood pressure was also measured at specified time intervals and correlated to enalaprilat plasma concentrations. Results: For enalapril, the AUCofi¥ values (Mean ± SD) were 450AE0 ± 199AE5 and 479AE 6± 215AE6 ng h/mL, Cmax values were 313AE5 ± 139AE6 and 310AE1 ± 186AE6 ng/mL, Tmax values were 1AE06 ± 0AE30 h and 1AE13 ± 0AE22 h, and t1/2 ranged between 0AE 3t o 6AE 1h( 1AE 6±1 AE5) and 0AE40 to 5AE05 h (1AE 3±1 AE0), for the two brands. For enalaprilat, the AUCofi¥ values were 266AE9 ± 122AE7 and 255AE9 ± 121AE8 ng h/ml, Cmax values were 54AE 8±2 9AE5 and 57AE 2±2 9AE0 ng/mL, Tmax values were 4AE 6±1 AE 6h and 4AE 3±1 AE45 h, and t1/2 ranged between 1AE 1t o 10AE 5h( 4AE 5±2 AE9) and 0AE 6t o 9AE 4h( 3AE 5±2 AE5) for the two brands. Conclusions: Cmax values for enalapril are about 10 times those published in the literature and the rate and extent of absorption of the two brands of enalapril and their deesterification to enalaprilat following the administration of either brand were bioequivalent. Secondly, enalaprilat concentrations at 12‐24 h following a single oral dose of enalapril in healthy volunteers were lower than those reported in the literature. The values reported here correlated with the return of blood pressure to predose level. Thirdly, enzyme immunoassays for enalapril and enalaprilat are better than ACE inhibition assays and can be used in bioequivalence assessment of enalapril and enalaprilat and for therapeutic drug monitoring in a clinical laboratory setting.

Estradiol, progesterone, prolactin and cortisol concentrations are substantially increased during pregnancy. Also, cortisol and prolactin levels are elevated during stress. In the present study, we exposed peripheral blood to estradiol, progesterone, prolactin and cortisol alone or in combination for 24 h before stimulation with T-dependent (phytohemagglutinin, PHA) and independent activators (lipopolysaccharide, LPS) to study their immunomodulatory role in interleukin-12 (IL-12), tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), and IL-10 production in a whole blood model. This should be similar to in vivo exposure conditions such as long-term stress, preovulatory or pregnancy periods. The present study showed that the stress-induced and preovulatory levels of prolactin and estradiol, respectively, increased the production of IFN-gamma and IL-12 levels (and IL-10 in the case of estradiol) in PHA + LPS-stimulated whole blood, and inhibited a hydrocortisone (100 nmol/l) suppressive effect on IFN-gamma, IL-12 and IL-10 productions. In LPS-stimulated whole blood, however, prolactin enhanced only IL-10 production levels in a non-concentration-dependent manner. Higher prolactin levels as in pregnancy did not modulate any of the cytokines, but pregnancy estradiol concentrations only induced higher IL-10 levels in PHA + LPS-stimulated whole blood. All progesterone levels tested revealed no effect on any of the cytokines following whole blood stimulation. Our results indicate that (1) a long exposure time of prolactin and estradiol to whole blood modulates the production of cytokines in a concentration- and stimulus-dependent manner; (2) stress-induced levels of prolactin and preovulatory estradiol concentrations can regulate cortisol-induced cytokine suppression, and (3) even though the cytokine pattern is different, pregnancy estradiol and cortisol levels decreased the IFN-gamma/IL-10 ratio, thereby keeping the anti-inflammatory IL-10 levels favored during pregnancy, which could be useful in regulating inflammatory-mediated autoimmune diseases.

Prolactin, an anterior pituitary hormone, has been shown to have a role in immunomodulation. Some reports have shown the importance of prolactin in activating lymphocytes and macrophages, while in hyperprolactinemia patients, prolactin was found to decrease lymphocyte activation and natural killer function. In the present work, at physiological (15ng/ml) and stress-induced levels (30ng/ml) of prolactin, interferon-gamma (IFN-gamma) and interleukin (IL)-12 p70 levels, but not of IL-10 and tumor necrosis factor-alpha (TNF-alpha), increased significantly (p

We have developed two enzyme linked immunosorbent assay (ELISA) methods for determining enalapril and enalaprilat in plasma. In this study, 48 healthy subjects received an oral dose of either 10 or 20 mg of enalapril and plasma concentrations of enalapril and enalaprilat were determined by their specific ELISA methods. These plasma concentrations and blood pressure measurements were applied to evaluate the pharmacokinetic (PK) and pharmacodynamic (PD) parameters of both enalapril and enalaprilat. The enalapril values for the area under the curve (AUC(0)--> infinity ) were 480 +/- 216 and 832 +/- 325 ngh/mL, maximum plasma concentrations (C(max)) were 310 +/- 187 and 481 +/- 185 ng/mL, and times required to reach the maximum concentration t(max) were 1.13 +/- 0.22 and 1.09 +/- 0.33 h for 10 and 20 mg doses, respectively. The enalaprilat values for AUC(0)--> infinity were 256 +/- 122 and 383 +/- 158 ngh/mL, C(max) values were 57 +/- 29 and 72.9 +/- 33.6 ng/mL and t(max) values were 4.28 +/- 1.45 and 4.05 +/- 01.22 h for 10 and 20 mg doses, respectively. The C(max) values of enalapril were approximately 10 times higher than those in the literature, which were determined by angiotensin converting enzyme (ACE) inhibition assays following alkaline hydrolysis, but similar to those of enalaprilat. The PD profiles revealed a significant correlation between enalaprilat concentrations in plasma and the decrease in systolic and diastolic blood pressures (r = -0.95 with P < 0.001 and r = -0.95 with P < 0.001), respectively, following a single oral dose of enalapril. These ELISA methods have the advantage of being simple, accurate, sensitive, and do not depend on enalaprilat binding to ACE. Such methods can be used for analysis and kinetic testing of enalapril and enalaprilat in biological fluids.

Paracetamol has an extensive first-pass metabolism that highly affects its bioavailability (BA); thus, dose may be repeated several times a day in order to have longer efficacy. However, hepatotoxicity may arise because of paracetamol metabolism. Therefore, this project aimed to increase paracetamol BA in rats by glucosamine (GlcN). At GlcN-paracetamol racemic mixture ratio of 4:1 and paracetamol dose of 10 mg/kg, paracetamol area under the curve (AUC) and maximum concentration (Cmax ) were significantly increased by 99% and 66%, respectively (p < 0.05). Furthermore, paracetamol AUC and Cmax levels were increased by 165% and 88% in rats prefed with GlcN for 2 days (p < 0.001). Moreover, GlcN significantly reduced phase Ι and phase I/ΙΙ metabolic reactions in liver homogenate by 48% and 54%, respectively. Furthermore, GlcN molecule was found to possess a good in silico binding mode into the CYP2E1 active site-forming bidentate hydrogen bonding with the Thr303 side chain. Finally, serum ALT and AST levels of rats-administered high doses of paracetamol were significantly reduced when rats were prefed with GlcN (p < 0.01). In conclusion, GlcN can increase the relative BA of paracetamol through reducing its metabolism. This phenomenon is associated with reduction in hepatocytes injury following ingestion of high doses of paracetamol.

This study evaluated the effects of a common stressful event, academic examination, on (1) reactivation of latent Epstein-Barr virus (EBV) and cytomegalovirus (CMV) and (2) levels of cortisol and prolactin in serum. Blood samples were drawn from 2 groups of female students at the beginning of the semester (baseline) and during final examination periods (stress samples) of summer and fall-winter semesters. During summer, fall-winter, and overall, examination stress resulted in significantly increased levels of cortisol at P = .037, .016 and .004, respectively, but not prolactin ( P = .32, .09 and .1, respectively). Examination stress significantly increased EBV viral capsid antigen (VCA) IgG levels from baseline levels ( P = .0028) in the summer group, but not in the fall-winter group. In addition, the difference in frequency of reactivation in EBV-seropositive subjects in the summer vs the fall-winter (64% vs 15%) was significant at P = .005. Furthermore, after the examination, the percent increase of EBV VCA IgG in the summer (56%) was significantly different ( P = .002) from the percent increase (–2%) in the fall-winter. On the other hand, examination stress did not affect the frequency of CMV reactivation or CMV IgG levels in either group. These data suggest that academic stress can reactivate some of the latent herpesviruses to various degrees at different seasons, and that in vivo reactivation of specific latent viral genes might involve multiple neuroendocrine interactions.

To study the effects of academic examination on Cortisol and prolactin, total leukocyte count, neutrophil counts, lymphocytes, and monocytes, blood samples were drawn from 58 female, fasting students at the beginning (baseline samples) and during final examination periods (stress samples) of summer or fall-winter semesters. Differences between baseline and stress samples were nonsignificant for total leukocytes (P = .9), and relative and absolute monocytes (P = .14 and .88, respectively). Stress samples showed significantly higher relative and absolute neutrophil counts. (P = .0035 and .033) and significantly lower relative and absolute lymphocyte counts (P = .007 and .00022). Cortisol levels were higher in the stress samples (P = .017). Overall prolactin level differences were not statistically significant. These data suggest that academic stress can modulate trafficking of lymphocytes and neutrophils but not monocytes. This redistribution may modulate the performance of the effector function of the immune system.

The combination of cytotoxic drugs with immunotherapy should be more effective than monotherapy alone since both therapeutic modalities may target different mechanisms. In addition, combination therapy may reduce adverse events associated with cytotoxic drugs. Eriobotrya japonica hydrophilic butanol-treated extract (EJWR) was found to modulate cytokines by enhancing IL-12, IFN-γ and TNF-α in vitro and in vivo and within tumor microenvironment. This was associated with enhancing survival time of mice bearing intra-peritoneal MCA fibrosarcoma (MCA FS). In the present work, we evaluated the combination of EJWR with doxorubicin (Dox) on MCA FS cytotoxicity using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction assay in the absence and presence of spleen cells or Natural Killer (NK) lymphocytes from tumor bearing mice. The results showed that Dox exhibited mild cytotoxicity to healthy spleen cells and EJWR reversed such cytotoxicity. In addition, increasing concentrations of Dox induced 40% (p

The successful use of herbal combinations in managing diseases or conditions over a single herb has lead us to evaluate the anti-dyslipidemic properties of the combination of the artichoke leaves extract, turmeric extract, prickly pear dried leaves (PPL) and garlic extract versus each one alone in two different hyperlipidemic animal models. A two-week treatment of each of the natural extracts, combination 1 (artichoke, turmeric and PPL) or combination 2 (artichoke, turmeric, PPL and garlic) prior to a single intraperitoneal injection of Pluronic F-127 resulted in decreasing significantly serum LDL levels by garlic and PPL extracts and serum LDL/HDL ratios by turmeric, PPL, combination 1 and 2. In a 10-day high fat diet model, only the combination 1 and 2 lowered serum cholesterol, LDL by 8–12%, decreased significantly triglycerides, LDL/HDL ratio; and increased significantly HDL (P<0.0001). However, a long term treatment of each natural product for 7 weeks resulted in decreasing significantly serum LDL levels and LDL/HDL ratio (P<0.05–0.0001). Furthermore, only artichoke and PPL inhibited significantly HMG-CoA reductase activity (P<0.05). In conclusion, short term, as well as long term, treatment using the combination of artichoke, turmeric, PPL and garlic extract prevents dyslipidemia; partially through inhibiting HMG-CoA reductase.

We recently designed a series of N-[4-(t-amino-yl)-but-2-yn-1-yl] isoindoline-1,3-diones as anti-inflammatory compounds, called ZM compounds. These ZM compounds were categorized according to the nature of the cyclic amino groups into ZM2, ZM3, ZM4, and ZM5 and were shown to reduce carrageenan-induced inflammation, inhibit cyclooxygenase (1 and 2) and have less adverse effects than the common non-steroidal anti-inflammatory drugs. In the present study, we are examining the potential effects of ZM compounds in modulating cytokines production in vivo and in vitro from stimulated spleen cells, CD4+ CD25+ve T regulatory cells and CD4+CD25-ve T helper cells. Six hours following oral administration of 20mg/kg of ZM4 and ZM5 compounds reversed LPS-induced TGF-β suppression whereas ZM2, ZM3, ZM4, and ZM5 reversed LPS-induced TNF-α and IL-12 increase in mice spleen. In addition, increasing concentrations of ZM2, ZM4 and ZM5 increased significantly TGF-β1 production, whereas ZM3, ZM4 and ZM5 suppressed only TNF-α production in LPS and LPS+PMA stimulated spleen cells. Furthermore, only ZM5, enhanced significantly TGF-β1 production from LPS and LPS+PMA stimulated CD4+CD25+ve cells (p

We have developed a series of aminoacetylenic isoindoline-1,3-dione compounds and showed their anti-inflammatory activities by reducing carrageenan-induced rat paw edema and modulating proinflammatory and anti-inflammatory cytokines. In the present study and due to efficacy reasons, we are exploring only two of these compounds, namely, ZM4 and ZM5, to reveal their analgesic activity and toxicity. Following oral administration, both compounds were effective in reducing significantly (P<0.05–0.001) acetic acid-induced writhing behavior, hot plate latency test, and formalin-induced paw licking time as antinociceptive indicators in mice and rats, respectively. Regarding the toxicity, the acute (20, 50, and 150 mg/kg) and repeated oral administration (10, 20, and 50 mg/kg) of these compounds for ten days did not produce any mortality and the compounds were considered well tolerated. However, repeated oral administration of 50 mg/kg of both compounds induced erythropoiesis by means of increasing significantly red blood cells, hemoglobin, and packed cell volume. Moreover, these compounds did not induce gastric lesions in the stomach of experimental animals at the doses that exhibited analgesic and anti-inflammatory activity compared to indomethacin as a positive control. The results indicate that ZM4 and ZM5 possess potential analgesic activity while being preliminarily safe and have minimal ulcerogenic activity.

Background Vitamin D is cutaneously synthesized following sun exposure (vitamin D3) as well as it is derived from dietary intake (vitamin D3 and D2). Vitamin D2 and D3 are metabolized in the liver to 25-hydroxyvitamin D (25(OH)D). This metabolite is considered the functional indicator of vitamin D stores in humans. Since Jordan latitude is 31°N, cutaneous synthesis of vitamin D3 should be sufficient all year round. However, many indications reveal that it is not the case. Thus, this study was conducted to determine the 25(OH)D status among Jordanians. Methods Three hundred healthy volunteers were enrolled in a cross sectional study; 201 females and 99 males. 25(OH)D and calcium concentrations were measured by enzyme linked immunosorbent assay and spectroscopy techniques, respectively. All participants filled a study questionnaire that covered age, sex, height, weight, diet, and dress style for females. Females were divided according to their dress style: Western style, Hijab (all body parts are covered except the face and hands), and Niqab (all body parts are covered including face and hands). Results The average plasma 25(OH)D levels in males and females were 44.5 ± 10.0 nmol/l and 31.1 ± 12.0 nmol/l, respectively. However, when female 25(OH)D levels were categorized according to dress styles, the averages became 40.3, 31.3 and 28.5 nmol/l for the Western style, Hijab and Niqab groups, respectively. These 25(OH)D levels were significantly less than those of males (p < 0.05, 0.001, 0.001, respectively). In addition, the plasma 25(OH)D levels of the Western style group was significantly higher than those of Hijab and Niqab groups (p < 0.001). Furthermore, dairy consumption in males was a positive significant factor in vitamin D status. Even though calcium concentrations were within the reference range, the Hijab and Niqab-dressed females have significantly less plasma calcium levels than males (p < 0.01). Conclusions Very low plasma 25(OH)D levels in females wearing Hijab or Niqab are highly attributed to low sunlight or UVB exposure. In addition, most of males (76%) and Western style dressed females (90%) have 25(OH)D concentrations below the international recommended values (50 nmol/l), suggesting that although sun exposure should be enough, other factors do play a role in these low concentrations. These findings emphasize the importance of vitamin D supplementation especially among conservatively dressed females, and determining if single nucleotide polymorphisms of the genes involved in vitamin D metabolism do exist among Jordanians.

Multiple innate and adaptive immune effector cells and molecules partake in the recognition and destruction of cancer cells to protect against growing tumors, a concept that is known as cancer immunosurveillance. Unfortunately, cancer cells are capable of avoiding this process by immunoselection of poorly immunogenic tumor cells variants along with subversion of the immune system and thus shaping both the tumor and its microenvironment. Cytokines represent part of the complex pattern of the immune response which can assist the development of cancer as well as to eliminate it. Simultaneously, a large number of cytokines may be involved in the complex interactions between host and tumor cells where this dynamic cross-talk, between tumors and the immune system, can either regulate tumor growth or tumor growth, invasion and metastasis take place. In this review, we are stressing on the interface between infiltrated immune cells and tumor cells with the emphasis on the bidirectional activities of specific cytokines: IFN-γ, TGF-β and IL-17 within the tumor microenvironment and their role in shaping it. In addition, the significance of modulating such cytokines in favor of anti-tumor response is discussed and merits the use of mixture of targeted modulators to overcome the network complexity of cytokines in the tumor microenvironment.

Dopamine (DA) administration in sepsis is used to modulate the hypotensive condition and to normalize the blood vessels perfusion. However, whether this administration of DA has an effect on the release of cytokines in vivo deserves investigation.Pre-exposure of DA (1 µg/ml) to whole blood enhanced IL-10 (30%) production level following LPS stimulation. This IL-10 enhancement became statistically significant (p0.001) upon the addition of D2-DA receptor (DAR) antagonists, Clozapine or Haloperidol. Furthermore, systemic administration of DA (0.5-50 mg/kg) in mice suppressed significantly LPS-induced TNF-α levels in blood, liver, spleen, brain, and lungs; IL-10 levels in blood, brain and liver; and IFN-γ levels in blood, liver, brain, and lungs. On the other hand, DA enhanced significantly LPS-induced IL-10 production in the lungs and spleen, and IFN-γ levels in the spleen. Administration of Clozapine (54 mg/kg) or Haloperidol (62 mg/kg) with LPS (1 µg) and DA (5 mg/kg) reversed DA suppressive effects on LPS-induced cytokines in blood, IFN-γ in brain and lungs, and enhanced significantly LPS-induced IL-10 production in blood, spleen, liver, and lungs.These results indicate that DA modulatory effect on LPS-induced blood cytokines-producing cells is mediated mainly by D2-DAR (D2/ D3/D4) through enhancing immune cells migration and extravasation into tissues. Furthermore, DA selectivity on cytokines modulation is tissue specific, mediated by the type of DAR expressed and on the immune cells lodged in each tissue.

We present herein a new herbal combination called Etana that is composed of five herbal extracts including Panax quinquelotius (Ginseng), Eurycoma longifolia (Tongkat Ali), Epimedium grandiflorum (Horny goat weed), Centella asiatica (Gotu Kola) and flower pollen extracts. Most of the above-mentioned extracts have a long historical and traditional use for erectile dysfunction (ED). On the basis of the mechanism of action of each of the above, a combination is introduced to overcome several physiological or induced factors of ED. This study was conducted to show an enhancement of erectile function in male rats. The animals were observed for 3 h after each administration for penile erection, genital grooming and copulation mounting, and the penile erection index (PEI) was calculated. The maximum response was observed at the concentration of 7.5 mg kg(-1) of Etana. At a 7.5 mg kg(-1) single dose, the percentage of responding rats was 53+/-7 with a PEI of 337+/-72 compared with 17+/-6 with a PEI of 30+/-10 for control animals. This PEI was significantly (P

Cytokines induce the balance between inflammatory versus regulatory or antibody mediated reactions. So modulating the release of cytokines or inducing them by immunomodulating agents is an attractive mode for treating or help in treating several diseases such as autoimmune diseases. Eriobotrya japonica is a plant that is traditionally thought to have anti-inflammatory activities. Several compounds were isolated from the plant and showed distinctive biological effects. The purpose of this study was to determine the effects of epicatechin (EC) isolated from Eriobotrya Japonica on IL-6, IL-8, and IL-10 productions in whole blood stimulated with phytohemagglutinin (PHA)+lipopolysaccharide (LPS), and to examine if these cytokines are modulated through NFkappaB pathway.Sixteen healthy males and females volunteered in the study. Blood samples were drawn, diluted, and cultured for 24 h with different concentrations of EC and then PHA+LPS was added for another 24 h. The supernatant, then, was harvested and assayed for cytokines. In addition, mixing studies of EC and hydrocortisone were performed to examine the cytoplasmic and nuclear fractions of NFkB levels in association with cytokine production levels.Increasing concentrations of EC (1-100 microg/ml) in PHA+LPS stimulated whole blood cells culture suppressed significantly (p0.001) the production of IL-6 and IL-8. Moreover, increasing concentrations of EC modulated significantly the production of IL-10, as there was a significant increase in IL-10 level at 0.1, 1.0, and 10 microg/ml (p=0.058-0.004), while a significant decrease at a concentration of 100 microg/ml EC (p=0.037) was observed. In addition, an additive effect between EC and hydrocortisone (HC, 100 nmol/l) was seen in the production of IL-10, as there was a significant increase in IL-10 level (32%) compared with 27% for EC (10 microg/ml) and 19% HC. Furthermore, a significant decrease in cytoplasmic fractions of NFkappaB p65 level was found in samples containing EC 1, 10 microg/ml, but not in 100 microg/ml, when compared with control (p0.03). These latter changes were accompanied with a 29%, 67%, and 98% increase, respectively, of NFkappaB p65 in nuclear fractions compared to 24% reduction of NFkappaB p65 level in HC culture (p0.03).These results indicate that EC suppresses the production of pro-inflammatory cytokines, IL6 and IL-8, enhances the production of anti-inflammatory cytokine, IL-10, and stimulates NFkappaB p65 translocation to nucleus in PHA+LPS stimulated whole blood culture.

Stimulating or modulating the release of cytokines by immunomodulators or immunostimulating agents is an attractive mode for treating several diseases such as viral infections. For instance, patients with viral infections may be in need of increasing or inducing T helper 1 (Th1) or proinflammatory cytokines, which ultimately activate T cytotoxic and Natural killer lymphocytes to kill virally infected cells. Of these agents, we found that Eriobotrya japonica hydrophilic leaf extract (EJHE) can induce and modulate cytokines in dose-dependent manner. Twenty-four hour exposure of increasing concentrations of EJHE increased significantly (p

This study aims to present the in vitro inhibitory effect of Psidium guajava and Juglans regia leaf extracts on the main developer of acne lesions, Propionibacterium acnes (P. acnes), and other organisms that are isolated from acne lesions. Thirty-eight subjects (males and females) who had various types of acne were enrolled in the study. The contents of the acne lesions were cultured and the frequency of P. acnes (alone and with Staphylococci spp.) was 47%, whereas the frequencies for Staphylococcus aureus and Staphylococcus epidermidis were 13% and 24%, respectively. The antimicrobial activities of Psidium guajava and Juglans regia leaf extracts, determined by disk diffusion method (zone of inhibition), were compared to tea tree oil (TTO), doxycycline and clindamycin antibiotics. The zones of inhibition due to the Psidium guajava and Juglans regia leaf extracts ranged from 15.8–17.6 mm against P. acnes, 11.3–15.7 mm against S. aureus and 12.9–15.5 mm against S. epidermidis, respectively. These zones of inhibition were significantly higher than those of TTO and equivalent in case of Staphylococci spp., but less in case of P. acnes, to those obtained from doxycycline or clindamycin. It can be concluded that Psidium guajava and Juglans regia leaf extracts may be beneficial in treating acne especially when they are known to have anti-inflammatory activities.

Determination of protein cytokines in local tissues would help to evaluate their local role in health, sickness behavior and immune-mediated diseases. Therefore, developing a simple quantitative method of protein cytokines in tissues/organs is highly important.Mouse tissues were collected following intraperitoneal administration of endotoxin-free PBS or lipopolysaccharide. A mild detergent, 0.1% Igepal, was added in a buffer to enhance cytokines extraction. The tissues were then disrupted, homogenized, centrifuged and the supernatants were collected and assayed using solid-phase immunoassays.The presence of 0.1% Igepal extracted significantly more TNF-alpha from liver (322%: p0.01), brain (358%: p0.05), lungs (1600%: p0.01), and more IL-10 from liver (220%: p0.001), brain (4650%: p0.001) than PBS alone. On the other hand, using 0.1% Igepal did not increase IFN-gamma extraction from liver, spleen, brain, lungs, skin and kidneys more than PBS alone. Furthermore, i.p. administration of LPS induced a differential milieu of cytokines. LPS increased significantly the production of TNF-alpha, IFN-gamma, and IL-10 from liver (521%, 123%, 72%: p0.01, 0.04, 0.04), brain (470%, 122%, 280%: p0.01, 0.03, 0.01), peritoneal lavage (p0.001) and blood (p0.001). However, the pattern of increase was different for the above cytokines in spleen, skin, lungs and kidneys.The extraction of protein cytokines from tissues was superior with addition of mild detergent. Furthermore, our results showed a differential cytokines response to LPS with respect to tissue and cytokine type. This method should provide an important tool for studying local protein cytokines in behavioral pattern, sickness behavior, and immune-mediated diseases as well as to determine local therapeutic efficacy of immunomodulatory drugs.

Previously, a large number of studies reported that psychological stress and psychiatric illness reduces immune responsiveness. However, it turned out that stress reduces immune responsiveness is an oversimplified statement because the interactions between central nervous system, endocrine system and the immune system are undoubtedly complex. Therefore, this study aims in reviewing mental stress models (e.g. brief and written examination stress as subacute and acute type of stressor) that have been utilized to understand the effect of stress on the neuroendocrine and immune systems.The published findings from human mental stress models on catecholamines, cortisol, prolactin levels and on T helper (Th) 1 and 2-induced cytokines are presented and discussed with respect to the in vitro and in vivo effects of glucocorticoids, catecholamines, and prolactin on the induction of cytokines.This review shows evidence that short-time (minutes) or preparation to a written examination, in those students who are stressed, induces the production of proinflammatory cytokines which may be related to Th1 response. However, longer mental stress (days) causes dysregulation in the immune function by shifting the cytokine response to Th2 response.The outcome from neuroendocrine and immune function prior to, following and after mental stress depends on multiple variables most importantly on the amount of stress, exposure time, coping behavior and adjustment of the individual. A few minutes of stress may improve immune performance but longer times of mental stress have detrimental effects that may lead to loss of immune integrity. Furthermore, studies on stress and common heath problems are necessary to increase our knowledge and understanding of the mechanisms responsible for producing neuroendocrine-induced immune changes in health and common diseases.

The purpose of this study was to determine the effects of estradiol and progesterone on interferon-gamma (IFN-gamma), interleukin (IL)-12, IL-10 and tumor necrosis factor-alpha (TNF-alpha) productions in polyclonal activators (phytohemagglutinin+lipopolysaccharide)-stimulated whole blood cultures.Nineteen healthy males and females volunteered in the study. Blood samples were drawn, diluted, and cultured for 24h with different concentrations of estradiol, progesterone or hydrocortisone and then PHA+LPS was added for another 24 h. The supernatant, then, was harvested and assayed for IL-12 p70, IFN-gamma, IL-10 and TNF-alpha.At preovulatory concentrations, estradiol enhanced significantly IFN-gamma, IL-12 and IL-10, but not TNF-alpha, production levels and reversed the suppressive effect of hydrocortisone in PHA+LPS stimulated whole blood. While IL-10 levels kept increasing at pregnancy estradiol concentrations, IFN-gamma, IL-12 levels and IFN-gamma/IL-10 ratio decreased significantly. No effect of progesterone on IL-12 p70, IFN-gamma, IL-10 and TNF production levels was observed.The present study shows that those pregnancy estradiol concentrations (and higher) enhance the production of IL-10 and reduce IL-12, IFN-gamma levels and IFN-gamma/IL-10 ratio in stimulated whole blood cells. Because of the known IL-10 inhibitory actions on T helper (Th) 1 cells and monocytes/macrophages, these high IL-10 levels keep Th2 cytokines favored during pregnancy and may be useful in shifting Th1-mediated autoimmune diseases towards non-pathogenic Th2 pathway.

Mujtaba M Badr,1,2 Nidal A Qinna,1,2 Fadi Qadan,2 Khalid Z Matalka1,2 1Department of Pharmacology and Biomedical Sciences, Faculty of Pharmacy and Medical Sciences, 2Petra University Pharmaceutical Center, University of Petra, Amman, Jordan Background: Dichloroacetate (DCA) is one of the new, promising anticancer drugs. DCA restores normal mitochondrial function and enables cancer cells to undergo apoptosis. In addition, DCA was found to modulate certain signaling pathways involving some transcription factors. The latter encouraged us to study DCA immunomodulatory activity on cytokines and their association with increasing DCA cancer cell cytotoxicity. Methods and results: Cell viability assay was used to determine the effect of different concentrations of DCA on the survival of 3-methylcholanthrene (MCA) fibrosarcoma cell line. DCA decreased the percent survival of MCA fibrosarcoma in a dose-dependent manner (P

Amlodipine is a calcium channel antagonist of the dihydropyridine group. It is effective for treating hypertension, chronic stable angina, and vasospastic angina. However, it is difficult clinically to pinpoint the maximum dosage for antihypertensive activity of the drug without having parallel data on the plasma drug concentrations. The methods for assaying amlodipine are either gas chromatography with electron capture detector or liquid chromatography coupled with tandem mass spectrometry (or with an electrochemical detector), which needs tedious derivatization, and is expensive and time consuming. Therefore, in this study we developed an enzyme immunoassay for determining amlodipine in plasma. Anti‐amlodipine antibodies were produced following immunization of bovine serum albumin‐amlodipine conjugate. These specific antibodies were used in a competitive biotin–avidin‐based enzyme‐linked immunosorbent assay to measure amlodipine in plasma. Biotin was linked to the antibodies in order to enhance the sensitivity of the assay. The assay was specific for the free form of amlodipine with a detection limit of 0.1 ng/ml and the intra‐ and interassay coefficient of variation ranged from 1.6–10.2%. This immunoassay provides a sensitive, reliable, rapid, and accurate method for determination of amlodipine in plasma, which can be used in therapeutic drug monitoring pharmacokinetic studies and pharmaceutical analysis. J. Clin. Lab. Anal. 15:47–53, 2001. © 2001 Wiley‐Liss, Inc.

Ferula harmonis, which is locally called 'zallouh' in the Middle East, is used as an aphrodisiac as it is reputed to enhance male sexual behavior, however, there is no scientific verification. In this study, the oil extracted from the seeds of Ferula harmonis was tested for its efficacy in enhancing erectile function and toxicity in male rats. The sexual activities assessed by penile erection index were dose dependent. The ED(50) (12.03 mg/kg) was 880 times less than the LD(50) (10.6 g/kg). However, when doses ranging from 0.05, 0.5 to 2 g/kg were given daily for 28 days, acute and subacute toxicity were observed. There was a decrease in total body weight, hepatomegaly, atrophic testis, significant decrease in hemoglobin and red blood cell count. In addition, there was a significant decrease in cholesterol level. All the above indicate that the crude oil from the plant Ferula harmonis can enhance erectile function, however, it becomes toxic if it is used for a long period of time. Further studies are underway to isolate and identify the active ingredients and their exact mechanisms of action.

Ethidium homodimer (EHD) was conjugated to B72.3 monoclonal antibody using a method whereby 85-90% of the conjugated EHD remains available for DNA intercalation. Antibody was thiopropionylated by reaction with N-succinimidyl 3-(2-pyridyldithio)propionate and reduction of pyridyldithio groups with dithiothreitol. EHD was maleimido-functionalized with succinimidyl-4-(N-maleimidoethyl)cyclohexane-1-carboxylate and treated with thiopropionylated antibody to obtain a conjugate containing approximately 3.4 EHD per antibody molecule. For biologic studies, 14C-labeled EHD was synthesized by reductive amination and conjugated as above. In vitro the conjugate maintained chemical integrity and immunoreactivity, while in vivo its targeting of LS174T tumors was reduced compared with that of iodinated antibody. A decrease in isoelectric point of the immunoconjugate was also observed.

Purpose: The present study was carried out to determine the efficacy of Boron Neutron Capture Therapy (BNCT) for intracerebral melanoma using nude rats, the human melanoma cell line MRA 27, and boronophenylalanine as the capture agent. Methods and Materials: Pharmacokinetic and tissue distribution studies: MRA 27 cells (2 X 10 5 ) were implanted intracerebrally, and 30 days later, 120 mg of 10 B-L-BPA were injected intraperitoneally into nude rats. Therapy experiments: Thirty days following implantation, tumor bearing rats were irradiated at the Brookhaven Medical Research Reactor. Results: Pharmacokinetic experiments: Six hours following administration of BPA, tumor, blood, and normal brain boron-10 levels were 23.7, 9.4, and 8.4 μ/g respectively. Therapy experiments: Median survival time of untreated rats was 44 days compared to 76 days and 93 days for those receiving physical doses of 2.73 Gy and 3.64 Gy, respectively. Rats that had received both 10 B-BPA and physical doses of 1.82, 2.73, or 3.64 Gy had median survival times of 170, 182, and 262 days, respectively. Forty percent of rats that had received the highest tumor dose (10.1 Gy) survived for > 300 days and in a replicate experiment 21% of the rats were longterm survivors (> 220 days). Animals that received 12 Gy in a single dose or 18 Gy fractionated (2 Gy X 9) of gamma photons from a 137Cs source had median survival times of 86 and 79 days, respectively, compared to 47 days for untreated animals. Histopathologic examination of the brains of longterm surviving rats, euthanized at 8 or 16 months following BNCT, showed no residual tumor, but dense accumulations of melanin laden macrophages and minimal gliosis were observed. Conclusion: Significant prolongations in median survival time were noted in nude rats with intracerebral human melanoma that had received BNCT thereby suggesting therapeutic efficacy. Large animal studies should be carried out to further assess BNCT of intracerebral melanoma before any human trials are contemplated.

Fifty percent of primary malignancies of the central nervous system are anaplastic astrocytomas and glioblastoma multiforme1. The prognosis of patients with these tumors is dismal despite aggressive surgery followed by radio-and chemotherapy, and characteristically the mean survival time of these patients still is approximately 12 months2,3.

Malignant melanoma is a highly metastatic tumor to the skin, liver, lung and especially the brain. Patients with brain metastases have a poor prognosis despite aggressive chemo-and radiotherapy. Many of these patients have disseminated melanoma but eventually succumb to their cerebral metastatic disease.