Search

The study utilizes Grice's (1975) conversational maxims (quality, quantity, relation, and manner) which make up the cooperative principle to investigate how and why Jordanians flout the maxims in their everyday conversations. There are many studies that examine speakers' non-observance of the conversational maxims in everyday conversations. However, this study aims to advance pragmatic theory by illuminating the new pragmatic strategies of Jordanian speakers' non-observance of maxims (flouting) and classifying them in accordance with these Grice's maxims for the purpose of developing a framework that can help researchers classify data into pragmatic strategies and linguistic forms based on flouting of these maxims and how this flouting functions. This study also aims to show how nonobservance constitutes an important factor in helping the addressee realize the intended meaning and maintain stronger relationships between Jordanian speakers as well. Furthermore, this study identifies the most prominent maxims that are flouted in such everyday conversations. Among the main findings is that Jordanian speakers flout maxims as an indication or hint for an implicature that the speaker makes for positive purposes that aim to maintain and strengthen social intimacy. Additionally, this study offers more in-depth explanations of the relationships between flouting's dimensions, drawing on micro, meso and macro concepts as one way to depict how implicature is made by the speaker and grasped by the hearer for securing social goals in Jordanian Arabic.

Background: Hyperglycemic metabolic disorders such as diabetes can impair gastrointestinal (GI) physiological functions leading to multiple digestive manifestations. Oxidative stress which is an imbalance between reactive oxygen species production and antioxidants is contributed to GI complications that occur during hyperglycemia. Aim: To investigate the effect of high-glucose concentration on the antioxidant enzymes in the GI tract. Methods: Small intestine and colon tissues extracted from rats were incubated in a high-glucose medium for 3 hours. Following tissue homogenization, antioxidant enzyme activity and expression were evaluated. Results: Catalase (CAT) activity was increased in the small intestine (1742 ± 113.1–2265 ± 242.4 [mU/mL]) and decreased in the colon (3791 ± 516.2–1532 ± 292.9). Total antioxidant capacity was decreased in the small intestine (10.1 ± 1.83–9.048 ± 0.441 [nmole/μl]) and increased in the colon (8.114 ± 0.9–11.01 ± 0.99). Messenger RNA (mRNA) expression of antioxidant enzymes in the small intestine was increased (CAT: 0.03941 ± 0.0041–0.1917 ± 0.0165, glutathione peroxidase 1 [GPx1]: 1.156 ± 0.0855–16.24 ± 1.618, glutathione reductase [GR]: 0.0413 ± 0.0014-0.1549 ± 0.0145, superoxide dismutase 1 [SOD1]: 1.03 ± 0.1095–8.52 ± 0.471, SOD2: 0.00106 ± 3.559e-005–0.0028 ± 0.00052, SOD3: 0.0352 ± 0.0044–0.0493 ± 0.0223). mRNA expression of antioxidant enzymes in the colon was decreased (CAT: 0.02148 ± 0.0032–0.01057 ± 0.0014, GPx1: 0.48 ± 0.146–0.1090 ± 0.0209, GPx4: 0.2391 ± 0.063–0.01671 ± 0.0019, GR: 0.0393 ± 0.0031–0.0093 ± 0.0014, SOD1: 0.389 ± 0.1159–0.088 ± 0.0251, SOD2: 0.000934 ± 0.00020–0.000233 ± 2.39023e-05, SOD3: 0.0114 ± 0.00107–0.0017 ± 0.000176). Conclusion: Most of the results indicate a state of oxidative stress in the GI tract mediated by the exposure to high glucose level. Diabetic GI complications could be reversed using specific modalities that act to increase the antioxidant capacity. [ABSTRACT FROM AUTHOR]

Background and objectives: Anemia and chronic kidney disease (CKD) are common findings in diabetic patients. Lack of glycemic control is associated with increased risk of diabetic complications. This study aimed to determine the effect of poor glycemic control on the prevalence and determinants of anemia and CKD among type 2 diabetes mellitus (T2DM) patients in Jordan. Methods: A cross-sectional study design was used in this research. T2DM patients with controlled diabetes (HbA1c ≤7.0%, n = 120) and age-, gender- and body mass index–matched uncontrolled diabetic patients (HbA1c >7.0%, n = 120) were recruited. Blood sample for HbA1c and serum insulin measurement were obtained. Complete blood count and kidney function test results were obtained from the patient's medical records. Anemia was determined according to World Health Organization criteria. A binomial logistic regression was performed to ascertain the effects of age, gender, CKD and glycemic control on the likelihood that participants have anemia. Results: The prevalence of anemia was significantly higher in the uncontrolled T2DM compared to controlled T2DM patients (40% vs 27.5%, OR: 2.14, 95% CI: 1.23, 3.71, P = 0.006). Female patients with uncontrolled T2DM had significantly greater prevalence of anemia compared to male patients with uncontrolled T2DM. The binomial logistic regression analysis showed that age, female gender, and CKD were positively associated with anemia in the multivariate model, while in the univariate model, lack of glycemic control increases the odds of anemia by 1.74 (95% CI: 1.01, 2.99, P = 0.046). Conclusion: Anemia is commonly present among T2DM patients in Jordan and is associated with poor glycemic control especially in females. These results emphasize the necessity of including anemia screening in standard diabetes care to enable early detection and treatment of anemia and to enhance the overall care of diabetic patients. [ABSTRACT FROM AUTHOR]

This study elucidates how speakers of different cultures, namely, Saudi Arabia and United States employ the speech act of requesting, drawing upon various politeness features. It conducted a qualitative analysis to offer explanations for different strategies of requests used by both groups and how they employ politeness features. The present study also conducted small statistics (frequencies and percentages) to explore the most and least common strategies that were used by participants in this study. The sample of this study included 20 Saudis and 20 Americans who were students at Southern Illinois University. The findings revealed that while Saudi participants used more direct strategies than their American counterparts, these direct strategies were characterized as positive politeness. That is, Saudi participants relied on directness feature to minimize the social distance with the addressee. However, American students were more formal than their Saudi counterparts, suggesting that they were less direct when they made requests. But this type of directness is negative as it doesn’t aim at removing social borders between interlocutors. This study indicates that Saudis tend to be more informal (and hence less indirect) than Americans when they make requests. The present study demonstrates that culture is a key social factor to determine how this speech act is employed by speakers of different cultures. It can thus be argued that directness has social implications as it plays an important role in eliminating social borders and minimizing social distance between the interlocutors. [ABSTRACT FROM AUTHOR]

The study investigates the impact of teaching vocabulary through semantic relations (i.e. including hyponymy, synonymy, antonymy, polysemy, homonymy and metonyms) on the breadth and depth of EFL learners’ vocabulary knowledge. A convenient sample of 50 freshman EFL learners from a state university in Jordan was recruited. The study followed an experimental approach; data were collected through pre- and post-tests over an academic term. An interview was also conducted to elicit students’ attitudes toward this vocabulary-learning strategy. The results of the ANOVA test have revealed that Jordanian EFL learners find certain difficulties concerning the identification and production of all semantic relations. They scored low in identifying semantic fields properly though they were more aware of synonyms and antonyms than others. The T-test analysis of the data revealed that the degree of awareness of the major types of semantic relations significantly differed between both tests. This implies that using this strategy has improved students' semantic awareness. Besides, students have shown positive attitudes towards using this technique asserting that it contributes to the development of their mental lexicon. The study has many pedagogical implications and recommendations for further research. [ABSTRACT FROM AUTHOR]

This study attempts to investigate the disagreement strategies that are used by American speakers of Arabic with a view to identifying which disagreement strategies they use in equal and non-equal status situations. In addition, it aims to see whether variables like gender and social status affect the linguistic choices and disagreement strategies that they use. The subjects of the study are 28 (14 male and 14 female) American speakers of Arabic who were learning Arabic and were residing in Jordan at the time of data collection. The researchers analyze their interactional recorded responses to a set of stimuli included in an oral (recorded) discourse completion task (ODCT) prepared for this purpose. The ODCT comprises six scenarios in which the respondent is requested to disagree with two peers, two higher-status interlocutors, and two lower-status interlocutors. The findings of the study show that the American speakers of Arabic use two main disagreement strategies, non-confrontational and confrontational disagreements, which are in turn divided into sub-strategies. Further, they employ the non-confrontational strategies slightly more than the confrontational ones, as the percentage for the former is 51% while for the latter is 49%. Interestingly, the study suggests that the topic of discussion significantly influences the choice of strategy, sometimes resulting in women being more confrontational than men, which contrasts with common perceptions reported in the literature about gender-based communication styles. [ABSTRACT FROM AUTHOR]

The use of language in digital discourse for marketing has rapidly developed through mass media. This paper elucidates how advertisers employ various pragmatic strategies to persuade the recipient to act (behavior) by purchasing specific products. This study utilized different theoretical and conceptual frameworks (Theory of Reasoned Action and Aristotle's Models of Persuasion) to address the shortcomings of the social cognitive approach in studying persuasion, to investigate how language of advertisements can influence the recipient's thinking of a product from a psychological perspective. Guided by the principles of TRA, the present study argues that persuasion in advertisements is structured by three dimensions: attraction (through language features and appeals), evaluation (through beliefs, attitudes, and intention), and behavior (social acceptance or reluctance). This study revealed eight persuasion techniques employed by advertisers including demonstrating distinction, honoring commitment, expressing authority, hyperbolizing, glorification, providing proofs, expressing solidarity, and proving success. Showing distinction and Honoring commitment were the most frequently used strategies. Additionally, the study found that strategies of persuasion involved ethical, logical, and emotional appeals for their large effect on the recipient as they contribute to the recipient's positive evaluations. Appealing to reasoning (logic) is the most common one in slogans. [ABSTRACT FROM AUTHOR]

This research examined how private speech affects the speaking ability competency in young Jordanian EFL (English as a Foreign Language) students. The mixed-methods research gathered quantitative and qualitative data from 50 Jordanian EFL students aged 10-12. Pre- and post-tests of speaking skill mastery provided quantitative data, and observations and semi-structured interviews were also conducted to provide qualitative data. Private speech exercises significantly improved participants' speaking skills. Private speech improved participants' fluency, accuracy, and complexity. The qualitative data also showed that private speech helped individuals speak English with confidence and less nervousness. The research reveals that private speech exercises might help young Jordanian EFL students improve their speaking skills. EFL instructors and curriculum designers in Jordan may want to include private speech exercises to improve students' speaking and language abilities. Private speech activities should be studied in additional circumstances and age ranges. [ABSTRACT FROM AUTHOR]

This paper investigates the speech act of making threats among native speakers of Jordanian Arabic (JA) and American English (AE). It explores new threat strategies used by Jordanian and American speakers and their pragmatic functions to construct an analytical framework for analyzing this act across cultures. The data for this study were collected using an open-ended questionnaire. This questionnaire consists of ten imaginary situations drawn from real life. The data were analyzed using chi-square tests (value <0.05) to determine whether the difference between the two groups for each threat strategy was statistically significant. The subjects of Jordanian Arabic included 40 male participants and 40 female participants from three universities in Irbid district while the American subjects included 15 male participants and 15 female participants from the University of Illinois in the United States. Five threat strategies were identified. Four of which were shared between the two groups: Telling Authority, Committing Harm, Introducing Options and Warning. However, Promise of Vague Consequence was confined to JA speakers. The study also found that JA speakers tended to be less direct than their AE counterparts. [ABSTRACT FROM AUTHOR]

The prevalence of type 2 diabetes mellitus (T2DM) is steadily increasing worldwide in an alarming fashion. Importantly, poor glycemic control is associated with development of various health sequalae's due to glucolipotoxicity, oxidative stress and increased inflammatory cytokines. The aim of the present study was to examine the effect of glycemic control on the relative abundance of inflammatory markers in patients with controlled and uncontrolled T2DM, and to test their association with the glycemic status in diabetic patients in Jordan. An observational cross-sectional study design was used. Patients with T2DM with controlled diabetes [glycated hemoglobin (HbA1c) ≤7.0%, n=110] and age-, sex- and body mass index (BMI)-matched uncontrolled diabetic patients (HbA1c >7.0%, n=105) were recruited. An antibody membrane array was used to examine the relative abundance of inflammatory cytokines and growth factors in the sera of the study subjects, followed by enzyme-linked immunosorbent assay (ELISA) to confirm the results. Fasting blood glucose, serum insulin, triglyceride and homeostatic model assessment for insulin resistance (HOMA-IR) score were significantly elevated in the uncontrolled T2DM group (P<0.05). Antibody membrane array showed that serum epidermal growth factor (EGF) is significantly decreased in the uncontrolled T2DM group, and this was confirmed by ELISA (158.77±111.7 vs. 95.9±82.7 pg/ml, P=0.002). The binary logistic model was used to predict the likelihood of being uncontrolled diabetic based on EGF levels. After controlling for age, sex and BMI, EGF was statistically associated with diabetes control, where lower EGF levels predicted uncontrolled diabetes. Additionally, Pearson's product-moment correlation showed a statistically significant negative correlation between EGF and HbA1c (r=-0.25, P<0.0001), and a positive correlation between HOMA-IR and HbA1c, (r=0.32, P<0.0001). The current data identify a novel link between serum EGF levels and the status of HbA1c indicative of diabetic control. [ABSTRACT FROM AUTHOR]

Transglutaminase (TG) isoforms control diverse normal and pathophysiologic processes through their capacity to cross-link extracellular matrix (ECM) proteins. Their functional and signalling roles in cardiac fibrosis remain poorly understood, despite some evidence of TG2 involvement in abnormal ECM remodelling in heart diseases. In this study, we investigated the role of TG1 and TG2 in mediating fibrotic signalling, collagen cross-linking, and cell proliferation in healthy fibroblasts by siRNA-mediated knockdown. siRNA for TG1, TG2 or negative control was transfected into cultured neonatal rat ventricular fibroblasts and cardiomyocytes. mRNA expression of TGs and profibrotic, proliferation and apoptotic markers was assessed by qPCR. Cell proliferation and soluble and insoluble collagen were determined by ELISA and LC-MS/MS, respectively. TG1 and TG2 were both expressed in neonatal rat cardiomyocytes and fibroblasts before transfection. Other TGs were not detected before and after transfection. TG2 was predominantly expressed and more effectively silenced than TG1. Knocking down TG1 or TG2 significantly modified profibrotic markers mRNA expression in fibroblasts, decreasing connective tissue growth factor (CTGF) and increasing transforming growth factor-β1 compared to the negative siRNA control. Reduced expression of collagen 3A1 was found upon TG1 knockdown, while TG2 knockdown raised α-smooth muscle actin expression. TG2 knockdown further increased fibroblast proliferation and the expression of proliferation marker cyclin D1. Lower insoluble collagen content and collagen cross-linking were evidenced upon silencing TG1 or TG2. Transcript levels of collagen 1A1, fibronectin 1, matrix metalloproteinase-2, cyclin E2, and BCL-2-associated X protein/B-cell lymphoma 2 ratio were strongly correlated with TG1 mRNA expression, whereas TG2 expression correlated strongly with CTGF mRNA abundance. These findings support a functional and signalling role for TG1 and TG2 from fibroblasts in regulating key processes underlying myocardial ECM homeostasis and dysregulation, suggesting that these isoforms could be potential and promising targets for the development of cardiac fibrosis therapies. [ABSTRACT FROM AUTHOR]

When comparing public and private schools, it is clear there are significant differences in teaching methods, and this may be used to help pupils become writers that are more creative. The goal of this research is to examine the concept of creative writing by analyzing comments made by EFL educators in their instruction. Fifteen EFL teachers in Jordan were interviewed and observed. The researcher interviewed fifteen teachers to get their opinions on how they think feedback may be used to foster creativity in writing among EFL students. The interview results revealed that thirteen instructors utilized student comments to encourage English as foreign language writers to be more creative. Moreover, observations found that ten instructors employed feedback in their courses, including peer feedback. This research supports the use of a pre- and post-test in an experimental evaluation of the impact of interventions (such as feedback) on students' writing creativity. Why some instructors oppose using creative writing exercises (feedback) is a subject that requires deeper exploration. [ABSTRACT FROM AUTHOR]

Oxytocin produces an excitatory effect on gastric muscle through the activation of receptors present on stomach smooth muscle cells. However, the intracellular mechanisms that mediate oxytocin excitatory effects are still largely unknown. Therefore, we aimed to investigate the signaling pathways involved in oxytocin-induced contractions in gastric smooth muscle, shedding light on phospholipase C (PLC)-β1 signaling and its downstream molecules, including inositol 1,4,5- trisphosphate (IP3) and myosin light chain kinase (MLCK). The contractions of gastric smooth muscle from male rats were measured in an organ bath set up in response to exogenous oxytocin 10−7 M, in the presence and absence of inhibitors of the indicated signaling molecules. Oxytocin (10−9–10−5 M) induced dose-dependent stomach smooth muscle contraction. Pre-incubation with atosiban, an oxytocin receptor inhibitor, abolished the oxytocin-induced contraction. Moreover, PLC β1 inhibitor (U73122) and IP3 inhibitor Xestospongin C inhibited oxytocin-induced muscle contraction to various degrees. Verapamil, a calcium channel blocker, inhibited oxytocin-induced contraction, and pre-incubation of the strips, with both verapamil and Xestospongin C, further inhibited the excitatory effect of oxytocin. Chelation of intracellular calcium with BAPT-AM (1,2-bis-(o-aminophenoxy) ethane-N,N,N′,N′-tetraacetic acid) significantly inhibited the effect of oxytocin on muscle contraction. Finally, pre-incubation of the strips with the Ca2+/calmodulin-dependent protein kinase selective inhibitor STO-609 significantly inhibited the contraction induced by oxytocin. These results suggest that oxytocin directly stimulates its cell surface receptor to activate PLC β1, which in turn liberates IP3, which eventually elevates intracellular calcium, the prerequisite for smooth muscle contraction. [ABSTRACT FROM AUTHOR]

This research paper investigates students' perceptions of whether English or the indigenous language (Arabic) should be used as a medium of instruction in English learning centers at the preintermediate level. This argument which is of both sides (Arabic or English should be used as a medium of instruction) has been investigated through students' responses. While there has been a great deal of research into students' perception of EMI in education, this study offers deeper explanations, from a psychological perspective, of why students prefer EMI despite the difficulty, which would negatively affect their attitudes toward English. In this study, the researcher employed a questionnaire survey to elicit some data from the participants studying in the EFL centers to present substantial evidence, represented by this study's findings, consulted, and supported by psychology-based explanations. Among the main findings is that although the participants of this study hold positive attitudes toward their native language (Arabic), they prefer to use English to Arabic as a medium of instruction. This study also found that awareness associated with business plays a key role in this preference. [ABSTRACT FROM AUTHOR]

Progesterone is a steroidal hormone that is produced from the corpus luteum of the ovaries and from the placenta. The main function of progesterone is to promote the secretory differentiation in the endometrium of the uterus and to maintain pregnancy by inhibiting uterine contractions throughout pregnancy. Progesterone performs its actions by activating the classical progesterone nuclear receptors that affect gene transcription and by the non-classical activation of cell surface membrane receptors that accounts for the rapid actions of progesterone. Besides the reproductive roles of progesterone, it exerts functions in many tissues and systems such as the nervous system, the bone, the vascular system, and the gastrointestinal (GI) tract. This review will summarize the recent literature that investigated the role of progesterone in GI tract motility. Most literature indicates that progesterone exerts an inhibitory role on gut smooth muscle cells in part by elevating nitric oxide synthesis, which induces relaxation in smooth muscle. Moreover, progesterone inhibits the signaling pathways that lead to contraction such as Rho kinase inhibition. These data serve as a quick resource for the future directions of progesterone research that could lead to better understanding and more effective treatment of gender-related GI tract motility disorders. [ABSTRACT FROM AUTHOR]

The production of pro-inflammatory cytokines and chemokines is increased during inflammatory bowel disease (IBD). Previously, it was demonstrated that brain derived neurotrophic factor (BDNF) expression is increased in experimental models of colitis. BDNF is partially responsible for the structural and functional changes that take place during IBD. However, the exact mechanisms underlying the upregulation of BDNF during gut inflammation are unknown. The aim of the present study was to determine the effects of direct treatment of smooth muscle cells with inflammatory cytokines on the synthesis and secretion of BDNF. BDNF expression and secretion levels were measured using ELISA kits on tissue lysates and on incubation media used to culture the rat colon smooth muscle tissues treated for 24 h with either tumor necrosis factor (TNF)-α or interleukin (IL)-1β. Compared with the control tissue samples, treatment with TNF-α and IL-1β resulted in a significant increase in the protein expression levels of BDNF in the incubated smooth muscle tissue. TNF-α and IL-1β also stimulated the secretion of BDNF. Chelation of intracellular Ca2+ with BABTA-AM prevented the TNF-α and IL-1β-induced increase in BDNF protein expression and secretion levels. Furthermore, inhibition of protein kinase A (PKA) significantly reduced BDNF expression levels when treated with cytokines but not secretion. In conclusion, proinflammatory cytokines that are upregulated during IBD, directly stimulated BDNF expression and secretion in a Ca2+ dependent manner. Considering the ability of BDNF to enhance smooth muscle contraction and pain sensation, this autocrine loop may partially explain the characteristic hypercontractility and hypersensitivity associated with IBD. [ABSTRACT FROM AUTHOR]

Background. Insomnia is a problem that is common in all societies and age groups. However, its importance is increasing between students especially with the highly competitive and demanding environment surrounding them even after their graduation. In spite of the deep understanding of its health and social consequences, the frequency of insomnia among medical students in Jordan was not determined. Aim. To determine the prevalence of sleep disturbances among college students and to look for any association between sleep disturbances and students' academic achievement. Methods. This is a cross-sectional self-administered questionnaire-based study. The participants were college students of the medical and paramedical specialties. Insomnia Severity Index (ISI) was used and the academic performance was assessed using students' Cumulative Grade Point Average (CGPA). Results. There were 977 responses. Prevalence of clinical insomnia was 26.0%. Students who self-reported good sleep quality had significantly lower ISI scores compared with those who self-reported bad quality of sleep. Students who slept >7 hours had significantly less ISI scores than students who slept <6 hours. Students who had a CGPA more than or equal to 3 had significantly lower ISI scores compared with those who had a CGPA less than 2.5. Self-reported sleep quality was associated with the CGPA. Conclusion. A high prevalence of insomnia was found in this group of students. Academic performance was significantly associated with ISI scores and self-reported sleep quality. These results might be useful for future research into the development of interventional strategies to help students get enough sleep quality and quantity. [ABSTRACT FROM AUTHOR]

Previous research has shown that smooth muscle of the stomach undergoes developmental changes in the intracellular regulatory mechanism responsible for the contractile process. Whether these developmental changes relate to differences in the expression and/or activity of the key enzymes regulating smooth muscle contraction has not been previously evaluated. Therefore, we aimed to examine the expression and activation of the small monomeric G protein "RhoA" and Rho kinase (ROCK) as well as their correlation with the contraction of gastric smooth muscle cells (GSMCs) in newborn vs. adult rats. Freshly isolated single GSMCs from Sprague-Dawley rats at 1 week (newborn) and 3 months (adult) of age were used in the study. Protein and mRNA expression levels of both ROCK2 and total RhoA were higher in adult compared to newborn rats. Moreover, acetylcholine (ACh)- induced contractions of GSMCs in adult rats were significantly higher than that in newborn animals. Meanwhile, ROCK and Rho activation was higher in adult stomach cells compared to newborn ones. Pretreatment of GSMCs with Y-27632, the ROCK inhibitor, significantly reduced ACh-induced contraction in both groups of cells and greatly abolished contractile differences. In conclusion, our results indicate that RhoA/ROCK pathway and contraction of stomach muscle cells are under developmental regulation. [ABSTRACT FROM AUTHOR]

It is well known that pregnancy is associated with frequent gastrointestinal (GI) disorders and symptoms. Moreover, previous reports have shown that estrogen, which changes in levels during pregnancy, participates in the regulation of GI motility and is involved in the pathogenesis of various functional disorders in the stomach. The aim of the current study was to explore the changes in the expression of estrogen receptors (ERs) and examine the effect of estrogen on nitric oxide- (NO-) cyclic guanosine monophosphate (cGMP) pathway and thus relaxation in gastric smooth muscle cells (GSMC) during pregnancy. Single GSMC from early-pregnant and late-pregnant Sprague-Dawley rats were used. Protein and mRNA expression levels of ERs were measured via specifically designed enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR), respectively. NO and cGMP levels were measured via specifically designed ELISA kits. Effect of estrogen on acetylcholine- (ACh-) induced contraction of single GSMC was measured via scanning micrometry in the presence or absence of the NO synthase inhibitor, N-nitro-L-arginine (L-NNA), or guanylyl cyclase inhibitor, 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ). Estrogen increased both NO and cGMP levels and their levels were greater in early compared to late pregnancy. Expression of ERs was greater in early compared to late pregnancy. ACh induced greater contraction of GSMC in late pregnancy compared to early pregnancy. Estrogen inhibited ACh-induced contraction in both periods of pregnancy. Importantly, pretreatment of GSMC with either L-NNA or ODQ abolished estrogen inhibitory action on muscle contraction. In conclusion, GSMC contractile behavior undergoes drastic changes in response to estrogen during pregnancy and this might explain some of the pregnancy-associated gastric disorders. [ABSTRACT FROM AUTHOR]

Previous studies have shown that progesterone could inhibit muscle contraction in various sites of the gastrointestinal tract. The underlying mechanisms responsible for these inhibitory effects of progesterone are not fully known. The aim of the current study was to investigate the effect of progesterone on the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) pathway and muscle contraction in the stomach. Single gastric smooth muscle cells from female Sprague-Dawley rats were used. The expression of progesterone receptor (PR) mRNA was analyzed by reverse transcription polymerase chain reaction. NO and cGMP levels were measured via specific ELISAs. Acetylcholine (ACh)-induced contraction of single gastric muscle cells preincubated with progesterone was measured via scanning micrometry in the presence or absence of the NO synthase inhibitor, Nω-Nitro-L-arginine (L-NNA), or guanylyl cyclase inhibitor, 1H-[1,2,4]Oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), and expressed as percent shortening from resting cell length. PR expression was detected in the stomach muscle cells. Progesterone inhibited ACh-induced gastric muscle cell contraction. Furthermore, progesterone increased NO and cGMP levels in single gastric muscle cells. Most notably, pre-incubation of muscle cells with either L-NNA or ODQ abolished the inhibitory action of progesterone on muscle contraction. These present observations suggest that progesterone promotes muscle cell relaxation in the stomach potentially via the NO/cGMP pathway. [ABSTRACT FROM AUTHOR]

Peroxynitrite is a reactive nitrogen species produced in the intravascular compartment from superoxide anion and nitric oxide. Peroxynitrite destroys blood plasma proteins and membranes of red blood cells and of platelets. This explains why excessive production of peroxynitrite contributes to diseases and to ageing. Therapeutics that antagonize peroxynitrite may delay ageing and the progression of disease. We developed an in vitro assay that allows the investigation of the oxidative damage caused by peroxynitrite in the intravascular compartment. This assay correlates the damage with the rate of formation of protein carbonyl groups, 3-nitrotyrosine (3-NT) and thiobarbituric acid reactive substances. Using this assay, we evaluated the ability of phenelzine, a scavenger of reactive aldehydes, to antagonize the effects of peroxynitrite. Herein, we showed that phenelzine significantly decreased the lipid peroxidative damage caused by peroxynitirite in blood plasma and platelets. Moreover, it inhibited carbonyl group and 3-NT formation in blood plasma and platelet proteins. [ABSTRACT FROM AUTHOR]

Peroxynitrite can be produced in the vasculature from a superoxide anion reaction with nitric oxide. A surplus of peroxynitrite in the intravascular compartment is a common feature of several chronic diseases. The development of pharmacological modalities that interfere with the formation of peroxynitrite or inhibit its oxidative damage may be of utility for the prevention and/or treatment of several pathologies. Our previous investigations showed that catalytically inactivating peroxynitrite-derived free radicals with tempol or scavenging reactive aldehyde species with phenelzine protects the blood plasma and platelets from the oxidative damage of peroxynitrite. However, the degree of inhibition of the cytotoxic effects of peroxynitrite using tempol or phenelzine was modest. In the present study, the aim was to examine if scavenging lipid peroxyl radicals with U-83836E can achieve superior protection from peroxynitrite. This was assessed by treating blood plasma or platelets with 100 µM peroxynitrite alone or in combination with U-83836E, and then measuring the levels of thiobarbituric acid reactive substances (TBARS) and protein carbonyl formation as indices of lipid peroxidation and protein oxidation, respectively. It was observed that scavenging lipid peroxyl radicals with 75–100 µM U-83836E increasingly reversed protein carbonylation induced by peroxynitrite in blood plasma and platelets, in addition to TBARS formation in blood plasma. These findings are further discussed in the context of the mechanisms by which U-83836E may protect against the cell-damaging effects of peroxynitrite. [ABSTRACT FROM AUTHOR]

Various gastrointestinal (GI) disorders have a higher prevalence in women than in men. In addition, estrogen has been demonstrated to have an inhibitory effect on the contractility of GI smooth muscle. Although increased plasma estrogen levels have been implicated in GI disorders, the role of gastric estrogen receptor (ER) in these sex-specific differences remains to be fully elucidated. The present study was designed to investigate the sex-associated differences in the expression of the two ER isoforms, ERα and ERβ, and the effect of estrogen on gastric muscle contraction via the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) pathway. Experiments were performed on single gastric smooth muscle cells (GSMCs) isolated from male and female Sprague Dawley rats. The effect of acetylcholine (ACh), a muscarinic agonist, on the contraction of GSMCs was measured via scanning micrometry in the presence or absence of 1 µM 17β-estradiol (E2), an agonist to the majority of ERs, 1,3,5-tris(4-hydroxyphenyl)-4-propyl-1H-pyrazole (PPT), an ERa agonist, or diarylpropionitrile (DPN), an ERβ agonist. The protein expression levels of ER subtypes in GSMCs were measured using a specifically designed ELISA. GSMCs from female rats had a higher expression of ERα and ERβ protein compared with GSMCs from males. ACh induced less contraction in female that in male GSMCs. Pre-treatment of GSMCs with E2 reduced the contraction of GSMCs from both sexes, but to a greater extent in those from females. PPT and DPN inhibited ACh-induced contraction in GSMCs from females. Furthermore, E2 increased NO and cGMP levels in GSMCs from males and females; however, higher levels were measured in females. Of note, pre-incubation of female GSMCs with Nω-nitro-L-arginine, a NO synthase inhibitor, or 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, a guanylyl cyclase inhibitor, reduced the inhibitory effect of estrogen on GSMC contraction. In conclusion, estrogen relaxes GSMCs via an NO/cGMP-dependent mechanism, and the reduced contraction in GSMCs from females by estrogen may be associated with the sex-associated increased expression of ERα and ERß, and greater production of NO and cGMP, compared with that in GSMCs from males. [ABSTRACT FROM AUTHOR]

Evidence of sex-related differences in gastrointestinal (GI) functions has been reported in the literature. In addition, various GI disorders have disproportionate prevalence between the sexes. An essential step in the initiation of smooth muscle contraction is the phosphorylation of the 20-kDa regulatory myosin light chain (MLC20) by the Ca2+/calmodulin-dependent myosin light chain kinase (MLCK). However, whether male stomach smooth muscle inherits different contractile signaling mechanisms for the regulation of MLC20 phosphorylation from that in females has not been established. The present study was designed to investigate sex-associated differences in the regulation of MLC20 phosphorylation and thus muscle contraction in gastric smooth muscle cells (GSMCs). Experiments were performed on GSMCs freshly isolated from male and female rats. Contraction of the GSMCs in response to acetylcholine (ACh), a muscarinic agonist, was measured via scanning micrometry in the presence or absence of the MLCK inhibitor, ML-7. Additionally, the protein levels of MLC20, MLCK and phosphorylated MLC20 were measured by ELISA. The protein levels of MLC20 and MLCK were indifferent between the sexes. ACh induced greater contraction (P<0.05) as well as greater MLC20 phosphorylation (P<0.05) in male GSMCs compared with female. Pretreatment of GSMCs with ML-7 significantly reduced the ACh-induced contraction (P<0.05) and MLC20 phosphorylation (P<0.05) in the male and female cells, and notably, abolished the contractile differences between the sexes. In conclusion, MLC20 phosphorylation and thus muscle contraction may be activated to a greater extent in male rat stomach compared with that in females. [ABSTRACT FROM AUTHOR]

Background: Intestinal smooth muscle cells (SMCs) undergo substantial morphological, phenotypic, and contractile changes during inflammatory bowel disease (IBD). SMCs act as a source and target for different inflammatory mediators, however their role in IBD pathogenesis is usually overlooked. Glucagon-like peptide-1 (GLP-1) is an incretin hormone reported to exert multiple anti-inflammatory effects in different tissues including the gastrointestinal tract through various mechanisms. Aim: The aim of this research is to explore the effect of GLP-1 analog exendin-4 on the expression and secretion of inflammatory markers from mouse colon smooth muscle cells (CSMCs) after stimulation with lipopolysaccharide (LPS). Materials and methods: Freshly isolated CSMCs from male BALB/c mice were cultured in DMEM and treated with vehicle, LPS (1 µg/mL), LPS+exendin-4 (50 nM), or LPS+exendin-4 (100 nM) for 24 h. Expression of inflammatory cytokines was then evaluated by antibody array membrane. Results: CSMCs showed basal expression of several cytokines which was enhanced with the induction of inflammation by LPS. However, exendin-4 (50 and 100 nM) significantly (p<0.05) reduced the expression of multiple cytokines including tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α), T cell activation gene-3 (TCA-3), stromal cell-derived factor-1 (SDF-1), and macrophage colony stimulating factor (M-CSF). To confirm these results, expression of these cytokines was further assessed by enzyme-linked immunosorbent assay and real-time polymerase chain reaction and similar results were also observed. Moreover, secretion of TNF-α and IL1-α into the conditioned media was significantly downregulated by exendin-4 when compared to LPS-treated cells. Furthermore, LPS increased NF-κB phosphorylation, while exendin-4 significantly reduced levels of NF-κB phosphorylation. Conclusion: These data indicate that GLP-1 analogs can exert significant anti-inflammatory effects on CSMCs and can potentially be used as an adjunct treatment for inflammatory bowel conditions. [ABSTRACT FROM AUTHOR]

Type 2 diabetes mellitus (T2DM) is a leading cause of death. The prevalence of T2DM in countries of the Middle East and North Africa (MENA) region, including Jordan, is among the highest worldwide. The reason(s) behind the epidemic nature of T2DM in Jordan are unknown but warrant further exploration. Studies have indicated that T2DM could be influenced by diet and/or genetic background. Evidence suggests that numerous patients with T2DM are deficient in vitamin D. The activity of vitamin D on its target tissues may be influenced by single nucleotide polymorphisms (SNPs) in the vitamin D receptor (VDR) gene. It was therefore hypothesized that SNPs in VDR could modify the risk of T2DM. To test this hypothesis, 125 patients with T2DM were recruited along with 125 controls. The study subjects were genotyped for variations in rs2228570, rs1544410, rs7975232, and rs731236 SNPs in the VDR. The levels of 25-hydroxyvitamin D [25(OH)D] were measured from the serum. The analysis revealed that reduced 25(OH)D and age were associated with the risk of T2DM (P<0.05). Moreover, under a dominant inheritance model, the GG genotype of rs2228570 was revealed to increase the risk of T2DM in univariate and multivariate analysis (P<0.05). Additionally, a chromosomal block containing the GAAG haplotype of VDR SNPs increased the risk of T2DM (OR=1.909; CI: 1.260-2.891; P=0.0021). Collectively, the present study revealed that low levels of serum 25(OH)D and rs2228570 of the VDR gene are associated with the risk of T2DM. [ABSTRACT FROM AUTHOR]

Recent studies have shown that both Rho kinase signaling and oxidative stress are involved in the pathogenesis of a number of human diseases, such as diabetes mellitus, hypertension, and atherosclerosis. However, very little is known about the effect of oxidative stress on the gastrointestinal (GI) smooth muscle Rho kinase pathway. The aim of the current study was to investigate the effect of oxidative stress on Rho kinase II and muscle contraction in rat stomach. The peroxynitrite donor 3-morpholinosydnonimine (SIN-1), hydrogen peroxide (H2O2), and peroxynitrite were used to induce oxidative stress. Rho kinase II expression and ACh-induced activity were measured in control and oxidant-treated cells via specifically designed enzyme-linked immunosorbent assay (ELISA) and activity assay kits, respectively. Single smooth muscle cell contraction was measured via scanning micrometry in the presence or absence of the Rho kinase blocker, Y-27632 dihydrochloride. All oxidant agents significantly increased ACh-induced Rho kinase II activity without affecting its expression level. Most important, oxidative stress induced by all three agents augmented ACh-stimulated muscle cell contraction, which was significantly inhibited by Y-27632. In conclusion, oxidative stress activates Rho kinase II and enhances contraction in rat gastric muscle, suggesting an important role in GI motility disorders associated with oxidative stress. [ABSTRACT FROM AUTHOR]

The signaling pathways mediating sustained contraction of mouse colonic longitudinal smooth muscle and the mechanisms involved in hypercontractility of this muscle layer in response to cytokines and TNBS-induced colitis have not been fully explored. In control longitudinal smooth muscle cells, ACh acting via m3 receptors activated sequentially Gα12, RhoGEF (LARG), and the RhoA/Rho kinase pathway. There was abundant expression of MYPT1, minimal expression of CPI-17, and a notable absence of a PKC/CPI-17 pathway. LARG expression was increased in longitudinal muscle cells isolated from muscle strips cultured for 24 h with IL-1β or TNF-α or obtained from the colon of TNBS-treated mice. The increase in LARG expression was accompanied by a significant increase in ACh-stimulated Rho kinase and ZIP kinase activities, and sustained muscle contraction. The increase in LARG expression, Rho kinase and ZIP kinase activities, and sustained muscle contraction was abolished in cells pretreated with the Jun kinase inhibitor, SP600125. Expression of the MLCP activator, telokin, and MLCP activity were also decreased in longitudinal muscle cells from TNBS-treated mice or from strips treated with IL-1β or TNF-α. In contrast, previous studies had shown that sustained contraction in circular smooth muscle is mediated by sequential activation of Gα13, p115RhoGEF, and dual RhoA-dependent pathways involving phosphorylation of MYPT1 and CPI-17. In colonic circular smooth muscle cells isolated from TNBS-treated mice or from strips treated with IL-1β or TNF-α, CPI-17 expression and sustained muscle contraction were decreased. The disparate changes in the two muscle layers contribute to intestinal dysmotility during inflammation. [ABSTRACT FROM AUTHOR]

Previous studies have identified differences in the expression of proteins that regulate myosin light chain phosphorylation and contraction in tonic and phasic smooth muscle cGMP plays a critical role in smooth muscle relaxation and is important for optimal function of phasic and tonic smooth muscle. The intracellular cGMP levels are regulated by its hydrolysis via phosphodiesterase 5 (PDE5) and efflux via novel multidrug resistance protein 5 (MRP5). In the present study we tested the hypothesis that the differences in the phasic and tonic behavior of smooth muscles may be related to differences in mechanisms that terminate cGMP signaling. Expression of PDE5 and MRP5 was significantly (more than 2-fold) higher in fundus compared with antrum. The NO donor S-nitrosoglutathione (GSNO) caused an increase in PDE5 activity and intra- and extracellular cGMP levels in both fundus and antrum. Stimulation of PDE5 activity and increase in extracellular cGMP were significantly higher in fundus, whereas increase in intracellular cGMP was significantly higher in antrum. GSNO-induced increase in extracellular cGMP was blocked in dispersed cells by the cyclic nucleotide export blocker probenecid and in cultured muscle cells by depletion of ATP or suppression of MRP5 by siRNA, providing evidence that cGMP efflux was mediated by ATPdependent export via MRP5. Consistent with the higher expression and activity levels of PDE5 and MRP5, GSNO-induced PKG activity and muscle relaxation were significantly lower in muscle cells from fundus compared with antrum. Thus higher expression of PDE5 and MRP5 in muscle cells from fundus correlates with tonic phenotype of muscle. [ABSTRACT FROM AUTHOR]

Caveolae act as scaffolding proteins for several G protein-coupled receptor signaling molecules to regulate their activity. Caveolin-1, the predominant isoform in smooth muscle, drives the formation of caveolae. The precise role of caveolin-1 and caveolae as scaffolds for G protein-coupled receptor signaling and contraction in gastrointestinal muscle is unclear. Thus the aim of this study was to examine the role of caveolin-1 in the regulation of Gq- and Gi-coupled receptor signaling. RT-PCR, Western blot, and radioligandbinding studies demonstrated the selective expression of M2 and M3 receptors in gastric smooth muscle cells. Carbachol (CCh) stimulated phosphatidylinositol (PI) hydrolysis, Rho kinase and zipper-interacting protein (ZIP) kinase activity, induced myosin phosphatase 1 (MYPT1) phosphorylation (at Thr696) and 20-kDa myosin light chain (MLC20) phosphorylation (at Ser19) and muscle contraction, and inhibited cAMP formation. Stimulation of PI hydrolysis, Rho kinase, and ZIP kinase activity, phosphorylation of MYPT1 and MLC20, and muscle contraction in response to CCh were attenuated by methyl β-cyclodextrin (MβCD) or caveolin-1 small interfering RNA (siRNA). Similar inhibition of PI hydrolysis, Rho kinase, and ZIP kinase activity and muscle contraction in response to CCh and gastric emptying in vivo was obtained in caveolin-1-knockout mice compared with wild-type mice. Agonist-induced internalization of M2, but not M3, receptors was blocked by MβCD or caveolin-1 siRNA. Stimulation of PI hydrolysis, Rho kinase, and ZIP kinase activities in response to other Gq-coupled receptor agonists such as histamine and substance P was also attenuated by MβCD or caveolin-1 siRNA. Taken together, these results suggest that caveolin-1 facilitates signaling by Gq-coupled receptors and contributes to enhanced smooth muscle function. [ABSTRACT FROM AUTHOR]

We examined expression of protease-activated receptors 2 (PAR2) and characterized their signaling pathways in rabbit gastric muscle cells. The PAR2 activating peptide SLIGRL (PAR2-AP) stimulated Gq, G13, Gi1, PI hydrolysis, and Rho kinase activity, and inhibited cAMP formation. Stimulation of PI hydrolysis was partly inhibited in cells expressing PAR2 siRNA, Gaq or Gai minigene and in cells treated with pertussis toxin, and augmented by expression of dominant negative regulator of G protein signaling (RGS4(N88S)). Stimulation of Rho kinase activity was abolished by PAR-2 or Ga13 siRNA, and by Ga13 minigene. PAR2-AP induced a biphasic contraction; initial contraction was selectively blocked by the inhibitor of PI hydrolysis (U73122) or MLC kinase (ML-9), whereas sustained contraction was selectively blocked by the Rho kinase inhibitor (Y27632). PAR2-AP induced phosphorylation of MLC20, MYPT1 but not CPI-17. PAR2-AP also caused a decrease in the association of NF-kB and PKA catalytic subunit: the effect of PAR2-AP was blocked by PAR2 siRNA or phosphorylation-deficient RhoA (RhoA(S188A)). PAR2-AP-induced degradation of IkBa and activation of NF-kB were abolished by the blockade of RhoA activity by Clostridium botulinum C3 exoenzyme suggesting RhoA-dependent activation of NF-kB. PAR2-AP-stimulated Rho kinase activity was significantly augmented by the inhibitors of PKA (myristoylated PKI), IKK2 (IKKIV) or NF-kB (MG132), and in cells expressing dominant negative mutants of IKK (IKK(K44A), IkBa (IkBa (S32A/S36A)) or RhoA(S188A), suggesting feedback inhibition of Rho kinase activity via PKA derived from NF-kB pathway. PAR2-AP induced phosphorylation of RhoA and the phosphorylation was attenuated in cells expressing phosphorylation-deficient RhoA(S188A). Our results identified signaling pathways activated by PAR2 to mediate smooth muscle contraction and a novel pathway for feedback inhibition of PAR2-stimulated RhoA. The pathway involves activation of the NF-kB to release catalytic subunit of PKA from its binding to IkBa and phosphorylation of RhoA at Ser188. [ABSTRACT FROM AUTHOR]

Gastrointestinal (GI) motility function and its regulation is a complex process involving collaboration and communication of multiple cell types such as enteric neurons, interstitial cells of Cajal (ICC), and smooth muscle cells. Recent advances in GI research made a better understanding of ICC function and their role in the GI tract, and studies based on different types of techniques have shown that ICC, as an integral part of the GI neuromuscular apparatus, transduce inputs from enteric motor neurons, generate intrinsic electrical rhythmicity in phasic smooth muscles, and have a mechanical sensation ability. Absence or improper function of these cells has been linked to some GI tract disorders. This paper provides a general overview of ICC; their discovery, subtypes, function, locations in the GI tract, and some disorders associated with their loss or disease, and highlights some controversial issues with regard to the importance of ICC in the GI tract. [ABSTRACT FROM AUTHOR]

Diabetes Mellitus (DM) currently ranks as the most common endocrine disorder worldwide. Current opinion views DM as a group of heterogeneous metabolic diseases characterized by hyperglycemia triggered by defects in the ability of the body to produce or use insulin in type 1 and 2 DM, respectively. Brain-derived neurotrophic factor (BDNF), one of the neurotrophin family of growth factors, has been linked to the pathogenesis of DM and insulin resistance. Moreover, vitamin D has been associated with insulin resistance and DM. Recently, the interactions between vitamin D and BDNF have been investigated in diabetic rats. However, this correlation has never been investigated in humans. Thus, the aim of the present study was to assess the alterations in serum BDNF and vitamin D levels in T2DM patients in Jordan, prior to and following vitamin D supplementation. A combination of non-experimental case-control and experimental designed studies were utilized to assess the relationship between serum BDNF and vitamin D levels in T2DM patients. The levels of BDNF and vitamin D were measured using commercially available ELISA kits, and fasting blood glucose (FBG) and HbA1c levels were measured in medical labs. The results showed that diabetic patients had lower levels of serum vitamin D and higher levels of BDNF compared with the healthy controls. Moreover, linear regression analysis indicated that BDNF levels were inversely correlated with serum vitamin D levels. Furthermore, vitamin D supplementation significantly increased vitamin D serum levels and decreased BDNF serum levels in diabetic patients. Intriguingly, FBG and HbA1c levels were significantly improved post vitamin D supplementation. These data demonstrate a positive effect of vitamin D supplementation in diabetic patients suggesting the implementation of vitamin D as part of future T2DM treatment plans. However, additional studies are needed to investigate the direct link between vitamin D, BDNF, and T2DM. [ABSTRACT FROM AUTHOR]

Type 2 diabetes mellitus (T2DM) is a worldwide health problem caused by resistance to insulin action. This chronic debilitating diseaseis preceded by a stage, known as prediabetes, in which a healthy lifestyle can delay the disease. The discovery of biochemical changes in prediabetes is important to identify individuals at risk of developing T2DM and in explaining disease pathogenesis. Adiponectin is secreted by fat cells and is linked with insulin resistance. Adiponectin levels are dysregulated in prediabetic subjects. This relationship had not been tested in Jordan. We recruited 130 subjects with prediabetes and 130 control subjects. We measured serum levels of adiponectin and genotyped subjects for three single nucleotide polymorphisms (SNPs) in the ADIPOQ gene; rs266729, rs1501299 and rs2241766. In multivariate analysis, we found that serum adiponectin lowers the risk of prediabetes (p = 0.002; odds ratio (OR), 0.764; 95% confidence interval (CI), 0.646–0.905). The rs1501299 SNP of the ADIPOQ gene was associated with prediabetes in our population (p = 0.041). Specifically, in multivariate analysis, the GT genotype of rs1501299 increased the risk of prediabetes (p = 0.010; OR, 2.350; 95% CI, 1.231–4.486) as well as the TT genotype (p = 0.006; OR, 4.774; 95% CI, 1.551–14.693). Our findings indicate that serum adiponectin and SNPs in the ADIPOQ gene are associated with prediabetes in Jordan. [ABSTRACT FROM AUTHOR]

Royal jelly (RJ) is a multifunctional bee product with a unique composition and wide-ranging biological properties, including antioxidant, anti-inflammatory and antiproliferative activities. Still, little is known about the possible myocardial protective properties of RJ. Considering that sonication could enhance RJ bioactivity, this study aimed to assess the effects of non-sonicated (NS) and sonicated (S) RJ on fibrotic signaling, cell proliferation, and collagen production in cardiac fibroblasts. S-RJ was produced by ultrasonication at 20 kHz. Ventricular fibroblasts isolated from neonatal rats were cultured and treated with different concentrations of NS-RJ or S-RJ (0, 50, 100, 150, 200, and 250 µg/well). S-RJ significantly depressed the expression levels of transglutaminase 2 (TG2) mRNA across all the concentrations tested and was inversely associated with the expression of this profibrotic marker. S-RJ and NS-RJ displayed distinct dose-dependent effects on mRNA expression of several other profibrotic, proliferation, and apoptotic markers. Unlike NS-RJ, S-RJ elicited strong negative dose-dependent relationships with the expression of profibrotic markers (TG2, COL1A1, COL3A1, FN1, CTGF, MMP-2, α-SMA, TGF-β1, CX43, periostin), as well as proliferation (CCND1) and apoptotic (BAX, BAX/BCL-2) markers, indicating that RJ dose-response effects were significantly modified by sonification. NS-RJ and S-RJ increased the content of soluble collagen, while decreasing collagen cross-linking. Collectively, these findings show that S-RJ has a greater range of action than NS-RJ for downregulating the expression of biomarkers associated with cardiac fibrosis. Reduced biomarker expression and collagen cross-linkages upon cardiac fibroblast treatment with specific concentrations of S-RJ or NS-RJ suggests putative roles and mechanisms by which RJ may confer some protection against cardiac fibrosis. • Royal jelly (RJ) displayed antifibrogenic activity in cultured ventricular fibroblasts. • RJ administration reduced collagen cross-linking in fibroblasts. • RJ altered the expression of key profibrotic, proliferation, and apoptotic markers. • Sonication enhanced RJ bioactivity in downregulating cardiac fibrosis signaling. • Sonicated RJ also showed superior potential for reducing fibroblast proliferation. [ABSTRACT FROM AUTHOR]