In vitro Quantification of Collagen and Snail1 Gene Expression in Experimentally Induced Fibrosis by Arecoline and Commercial Smokeless Tobacco Products.

INTRODUCTION: Extracellular matrix component derangement is the major event in pathogenesis of Oral submucous fibrosis. Many studies have elaborated the alteration of the matrix components at a cellular and genetic level. However elaborate quantification of the components with varying concentrations of Areca nut extract  and commercial tobacco products have not been done so far. MATERIALS AND METHODS: Primary culture of tissues sourced during crown lengthening procedures were used for establishment of fibroblast monoculture and fibroblast / keratinocyte co-culture. Extracts of areca nut, commercial smokeless tobacco products (gutkha and haans) and control CCl4 were tested at concentrations  ranging from 20 µL, 40 µL, 80 µL, 160 µL, 320 µL and time intervals of 12, 24, 48, 72 hours. Collagen quantification by spectrophotometry and SNAI1 gene expression study were done. RESULTS: Extract of areca nut was found to show increased collagen production than commercial tobacco products and closely similar values to CCL4. Kruskal Wallis test was used to analyse the difference in collagen obtained. The mean values of collagen obtained in co-culture were lesser than those obtained in the fibroblast monoculture. SNAI1 gene expression was negative in both the culture experiments. CONCLUSION: Areca nut extract was found to be more potent as an individual agent. Commercial smokeless tobacco products Gutka and Hans exhibited increased collagen production at higher concentration. These findings further steps up the persuasive ill effects of  tobacco products. Negative SNAI1 gene expression was corroborated to  lack of extracellular environment in the co coculture experiment.

The Dynamic Effects of Isosteviol on Insulin Secretion and Its Inability to Counteract the Impaired ß-Cell Function during Gluco-, Lipo-, and Aminoacidotoxicity: Studies In Vitro.

Isosteviol (ISV), a diterpene molecule, is an isomer of the backbone structure of a group of substances with proven antidiabetic capabilities. The aim of this study was to investigate if ISV elicits dynamic insulin release from pancreatic islets and concomitantly is able to ameliorate gluco-, lipo-, and aminoacidotoxicity in clonal ß-cell line (INS-1E) in relation to cell viability and insulin secretion. Isolated mice islets placed into perifusion chambers were perifused with 3.3 mM and 16.7 mM glucose with/without 10-7 M ISV. INS-1E cells were incubated for 72 h with either 30 mM glucose, 1 mM palmitate or 10 mM leucine with or without 10-7 M ISV. Cell viability was evaluated with a Cytotoxic Fluoro-test and insulin secretion was measured in Krebs-Ringer Buffer at 3.3 mM and 16.7 mM glucose. In the presence of 3.3 mM glucose, 10-7 M ISV did not change basal insulin secretion from perifused islets. However, at a high glucose level of 16.7 mM, 10-7 M ISV elicited a 2.5-fold increase (-ISV: 109.92 ± 18.64 ng/mL vs. +ISV: 280.15 ± 34.97 ng/mL; p < 0.01). After 72 h gluco-, lipo-, or aminoacidotoxicity in INS-1E cells, ISV treatment did not significantly affect cell viability (glucotoxicity, -ISV: 19.23 ± 0.83%, +ISV: 18.41 ± 0.90%; lipotoxicity, -ISV: 70.46 ± 3.15%, +ISV: 65.38 ± 2.81%; aminoacidotoxicity: -ISV: 8.12 ± 0.63%; +ISV: 7.75 ± 0.38%, all nonsignificant). ISV did not improve impaired insulin secretion (glucotoxicity, -ISV: 52.22 ± 2.90 ng/mL, +ISV: 47.24 ± 3.61 ng/mL; lipotoxicity, -ISV: 19.94 ± 4.10 ng/mL, +ISV: 22.12 ± 3.94 ng/mL; aminoacidotoxicity: -ISV: 32.13 ± 1.00 ng/mL; +ISV: 30.61 ± 1.54 ng/mL, all nonsignificant). In conclusion, ISV acutely stimulates insulin secretion at high but not at low glucose concentrations. However, ISV did not counteract cell viability or cell dysfunction during gluco-, lipo-, or aminoacidotoxicity in INS-1E cells.

Arecoline ameliorates hyperthyroid condition in mice under cold stress.

Betel nut of Areca catechu is chewed by millions of people for increased capacity to work and stress reduction, but it contains arecoline that causes hypothyroidism. The aim is to investigate the role of arecoline on thyroid activity in cold stress in mice. Arecoline treatment (10 mg/kg body wt/day, for 7 d) caused a reduction in thyroid weight and ultrastructural degeneration of thyro-follicular cells with depletion of T3 and T4 levels compared with the control mice. Cold stress (4 °C for 2 h, twice daily, for 7 d) stimulated thyroid activity ultrastructurally with an elevation of T3 and T4 levels. Arecoline treatment in cold stress suppressed thyroid activity by showing reversed changes to those of cold stress. In contrast, TSH concentrations were consistently increased under all experimental conditions. The findings suggest that cold stress causes hyperthyroidism which arecoline can ameliorate in mice.

Is Pilocarpine Effective in Preventing Radiation-Induced Xerostomia? A Systematic Review and Meta-analysis.

PURPOSE: To evaluate the efficacy of concomitant administration of pilocarpine on radiation-induced xerostomia in patients with head and neck cancers. METHODS AND MATERIALS: The PubMed, Web of Science, Cochrane Library, and ClinicalTrials were searched to identify randomized, controlled trials studying the effect of concomitant administration of pilocarpine for radiation-induced xerostomia. Included trials were systematically reviewed, and quantifiable outcomes were pooled for meta-analysis. Outcomes of interest included salivary flow, clinician-rated xerostomia grade, patient-reported xerostomia scoring, quality of life, and adverse effects. RESULTS: Six prospective, randomized, controlled trials in 8 articles were included in this systematic review. The total number of patients was 369 in the pilocarpine group and 367 in the control group. Concomitant administration of pilocarpine during radiation could increase the unstimulated salivary flow rate in a period of 3 to 6 months after treatment, and also reduce the clinician-rated xerostomia grade. Patient-reported xerostomia was not significantly impacted by pilocarpine in the initial 3 months but was superior at 6 months. No significant difference of stimulated salivary flow rate could be confirmed between the 2 arms. Adverse effects of pilocarpine were mild and tolerable. CONCLUSIONS: The concomitant administration of pilocarpine during radiation increases unstimulated salivary flow rate and reduces clinician-rated xerostomia grade after radiation. It also relieves patients' xerostomia at 6 months and possibly at 12 months. However, pilocarpine has no effect on stimulated salivary flow rate.

Arecoline induces TNF-alpha production and Zonula Occludens-1 redistribution in mouse Sertoli TM4 cells.

BACKGROUND: Arecoline, a major alkaloid in Areca nut has the ability to induce oxidative stress. The effect of Areca nut, arecoline on reducing sperm quality and quantity were documented previously using several animal models. Junction disruption by down-regulation of the junction-adhesive protein via oxidative stress is an important route mediating abnormal spermatogenesis. Therefore, in this present study, we investigated the functional role of arecoline on junctional proteins. RESULTS: To analyze direct effects of arecoline on testis cells, confluent mouse testicular Sertoli cell line TM4 was exposed to arecoline. Arecoline decreased insoluble zonula occludens-1 (ZO-1) protein expression in TM4 cells, however, arecoline treatment increased TNF-alpha production in both TM4 and monocytic THP1 cells. In addition, ERK1/2 inhibitor PD98059 reversed arecoline effects on TNF-alpha and ZO-1. CONCLUSIONS: Arecoline increases the production of TNF-alpha and induces protein redistribution of ZO-1. All these results explain the role of arecoline in male reproductive dysfunction, besides its cytotoxic induction.

Síndrome coronario agudo: una complicación grave e infrecuente en la deshabituación tabáquica con vareniclina / Acute coronary syndrome: An uncommon serious complication of smoking cessation with varenicline

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Tratamento da síndrome da bexiga hiperativa idiopática refratária / Treatment of idiopathic refractory overactive bladder syndrome

A síndrome da bexiga hiperativa é um distúrbio caracterizado pela presença de urgência miccional, podendo ou não haver incontinência urinária associada. Afeta homens e mulheres em iguais proporções, mas tem maior impacto sobre a população feminina, já que a incidência de incontinência é maior. O tratamento de primeira linha consiste em medidas comportamentais, treinamento vesical e fisioterapia, podendo-se associar tratamento farmacológico em casos persistentes. As drogas mais frequentemente utilizadas são os anticolinérgicos, medicações eficazes, porém associadas a efeitos adversos incômodos que frequentemente limitam a aderência. Uma proporção considerável de mulheres não obtém êxito com a combinação de medidas conservadoras e medicações anticolinérgicas, seja por eficácia limitada, seja por efeitos colaterais intoleráveis. Para essa população, modalidades de tratamento de segunda e terceira linha podem trazer alívio sintomático e melhora da qualidade de vida. Além disso, foi recentemente aprovada uma nova classe de drogas para o tratamento da bexiga hiperativa, os agonistas de receptores ?3-adrenérgicos, que prometem eficácia equivalente aos anticolinérgicos sem os efeitos adversos que os limitam...

Overactive bladder syndrome is a condition characterized by urgency, with or without urinary incontinence. It affects men and women equally, but has a greater impact on the female population, given the higher prevalence of urgency-incontinence. First line treatment consists on lifestyle interventions, bladder drills and physical therapy. Pharmacological treatment may be associated in persistent cases. The most commonly used medications are anticholinergics, which are efficacious, but limited by a variety of bothersome adverse effects that impair treatment compliance. A significant proportion of women don?t experience a successful outcome with the combination of conservative measures and treatment with anticholinergics, owing both to limited efficacy and to intolerable adverse effects that lead to treatment discontinuation. For this population, second and third line therapies may provide symptomatic relief, with great improvement in health related quality of life. Also, a new class of drugs, the ?3-adrenergic receptor agonists, has recently been approved for the treatment of overactive bladder, and may provide similar efficacy to currently used anticholinergic drugs without the associated adverse effects...

Lack of correlation between vasodilatation and pharmacologically induced immediate headache in healthy subjects.

BACKGROUND: The causal relationship between experimental headache and vasodilatation has not been fully clarified. In the present study, we combined headache and vascular data from eight experimental studies and conducted detailed statistical analyses. Given that substances used in all these experiments were vasodilators we examined a possible correlation between headache scores and increases in arterial diameter. METHODS: We identified nine studies and retrieved raw data in 89 healthy subjects (46 females, 43 males), mean age 27 years (range 18-59 years). The following variables were collected: maximal median headache intensity scores on a verbal rating scale (VRS) during immediate headache (0-120 minutes); the mean velocity of blood flow in the middle cerebral artery (V(meanMCA)); and the diameter of the frontal branch of the superficial temporal artery (STA) during the maximal median headache intensity. RESULTS: The scatter plots show no relationship between maximal headache score and the relative changes in V(meanMCA) and diameter of the STA. The main analyses of covariance showed a significant effect only of heart rate on headache (p = .014). The interaction tests were insignificant for all variables. CONCLUSIONS: The major outcome is a finding of no linear relationship between experimental immediate headache and dilatation of the MCA or STA.

Carbachol induces headache, but not migraine-like attacks, in patients with migraine without aura.

Carbachol induces headache in healthy subjects, but the migraine eliciting effect of carbachol has not previously been studied. We hypothesized that the cholinomimetic agonist carbachol would induce headache and migraine-like attacks in migraineurs. Carbachol (3 µg/kg) or placebo was randomly infused into 18 patients with migraine without aura in a double-blind crossover study. Headache was scored on a verbal rating scale from 0 to 10. Velocity in the middle cerebral artery (V(MCA)) and diameter of the superficial temporal artery (STA) were recorded. Fifteen patients experienced headache after carbachol compared with eight after placebo (P = 0.039). There was no difference in incidence of migraine-like attacks after carbachol (n = 8) compared with placebo (n = 6) (P = 0.687). Carbachol caused a decrease in V(MCA) (P = 0.044), but no change in STA (P = 0.089) compared with placebo. The study demonstrated that carbachol provocation is not a good model for experimental migraine.

Augmented heme oxygenase-1 expression in areca quid chewing-associated oral submucous fibrosis.

OBJECTIVES: Heme oxygenase-1 (HO-1) is known as a stress-inducible protein and functions as an antioxidant enzyme. HO-1 is consistently and dramatically upregulated in a variety of fibrotic diseases. The aim of this study was to compare HO-1 expression in normal human buccal mucosa and oral submucous fibrosis (OSF) specimens and further explore the potential mechanism that may lead to induce HO-1 expression. METHODS: Twenty OSF specimens and 10 normal buccal mucosa were examined by immunohistochemistry. The mRNA levels of HO-1 from fibroblasts cultured from OSF and normal buccal mucosa fibroblasts (BMFs) were evaluated by reverse transcription polymerase chain reaction. The effect of arecoline, the major areca nut alkaloid, was added to explore the potential mechanism that may lead to induce HO-1 expression. RESULTS: Heme oxygenase-1 expression was significantly higher in OSF specimens (P < 0.05) and expressed mainly by fibroblasts, endothelial cells, and inflammatory cells. OSF demonstrated significantly higher HO-1 mRNA expression than BMFs (P < 0.05). Arecoline was also found to elevate HO-1 mRNA and protein expression in a dose-dependent manner (P < 0.05). CONCLUSIONS: Taken together, the data presented here demonstrated that HO-1 expression is significantly upregulated in OSF from areca quid chewers, and arecoline may be responsible for the enhanced HO-1 expression in vivo.

Periodontal health status in pan chewers with or without the use of tobacco.

BACKGROUND: Betel nut and tobacco chewing is a common practice in south-east Asia. In India, betel nut is commonly chewed in the form of pan, with or without tobacco. Numerous studies have shown the carcinogenic potential of betel nut and tobacco. Betel nut and tobacco are also known to have deleterious effects on the oral tissues. PURPOSE: The aim of our study was to evaluate and compare the periodontal effects of pan chewing with or without the use of tobacco as an ingredient. MATERIALS AND METHODS: The periodontal status of 300 subjects (150 subjects were pan chewers with tobacco and 150 subjects were pan chewers without tobacco) was evaluated using the community periodontal index (CPI). The subjects were selected by the stratified random sampling method. The oral hygiene status of the subjects was evaluated using the simplified oral hygiene index. RESULTS: CPI code-4, with a probing depth of 6 mm or more, was seen in 30% of pan chewers with tobacco compared with 7.3% of pan chewers without tobacco. It was found that pan chewers with tobacco had 4.7 times more risk of having pockets than pan chewers without tobacco. The higher codes of loss of attachment were seen in pan chewers with tobacco compared with pan chewers without tobacco. It was found that pan chewers with tobacco had 7 times more risk of having loss of attachment when compared with the pan chewers without tobacco. CONCLUSIONS: The results show higher incidence of periodontal diseases in pan chewers who use tobacco compared with pan chewers who do not use tobacco. Based on the results, it was concluded that, although betel nut has deleterious effects on the periodontium, the addition of tobacco leads to a synergistic effect between betel nut and tobacco on the periodontal tissues.

The upregulation of cystatin C in oral submucous fibrosis.

Cystatin C is a 13kDa non-glycosylated basic protein belonging to cystatin family. It is consistently and dramatically upregulated in a variety of fibrotic diseases. The aim of this study was to compare cystatin C expression in normal human buccal mucosa and oral submucous fibrosis (OSF) specimens and further explore the potential mechanism that may lead to induction of cystatin C expression. Twenty-five OSF specimens and six of normal buccal mucosa were examined by immunohistochemistry. The activity of cystatin C from fibroblasts cultured from OSF and normal buccal mucosa were evaluated by using reverse-transcriptase polymerase chain reaction and enzyme-linked immunosorbent assay. Furthermore, the effect of arecoline, the major areca nut alkaloid, was explored. Cystatin C expression was significantly higher in OSF specimens (p<0.05) and expressed mainly by fibroblasts, endothelial cells, and inflammatory cells. OSF demonstrated significantly higher cystatin C expression than normal buccal mucosa fibroblasts both in mRNA and protein levels (p<0.05). In addition, arecoline was also found to elevate cystatin C mRNA and protein expression in a dose-dependent manner (p<0.05). Taken together, the data demonstrate that cystatin C expression is significantly upregulated in OSF from areca quid chewers and arecoline may be responsible for the enhanced cystatin C expression in vivo.

Prevención de los efectos adversos anticolinérgicos inducidos por los antipsicóticos en la 3ª edad: descripción de un caso en tratamiento con clozapina / Prevention of anticholinergic adverse effect induced by antipsychotics in aged: report of a case in treatment with clozapine

La seguridad y eficacia del uso de Clozapina en ancianos es actualmente objeto de estudio. Los pacientes geriátricos parecen presentar una mayor predisposición a sufrir hipotensión ortoestática y una mayor reactividad a los efectos anticolinérgicos, como también son más sensibles a los efectos adversos sobre el SNC. Se describe en el presente estudio el caso de una paciente portadora de un cuadro esquizofrénico catatónico en tratamiento con Clozapina, que presentó marcados síntomas anticolinérgicos y que mejoró de sus síntomas tras la suspensión del fármaco y el cambio a otro antipsicótico atípico (Risperidona). El estudio de niveles plasmáticos de Clozapina mostró que aún con 50 mg/día alcanzaba niveles plasmáticos de 200 ng/dl. Se analiza en el presente reporte la necesidad de una adecuada evaluación clínica y la importancia del control de Niveles Plasmáticos.

Glaucoma medications: use and safety in the elderly population.

As with many diseases, glaucoma increases in frequency in older populations, and is very often encountered in patients taking multiple medications. While the exact mechanism of glaucomatous optic neuropathy (GON) is not known, intraocular pressure (IOP) is thought to be central to the process, and reducing IOP is the only known effective treatment. The newer definition of glaucoma is an IOP-sensitive optic neuropathy. While large, controlled studies have indicated that reducing IOP will slow the progression of disease, the contributions of other conditions and medications have not been adequately studied. As the adverse effect profiles of medical therapies for glaucoma have improved, use of these agents has increased greatly. This has resulted in a large number of older patients taking glaucoma medications. Since topical medications can easily be overlooked in a medical history, and are for the most part well tolerated, systemic complications from these agents can be missed. In addition to being a common disease requiring treatment, glaucoma is also a model system for other degenerative diseases, and many of the concepts originally developed in relation to neurodegenerative diseases such as Alzheimer's disease are under investigation for glaucoma. These include approaches targeted towards neuroprotection and excitotoxicity.

Management of glaucoma: focus on pharmacological therapy.

Glaucoma represents a major cause of vision loss throughout the world. Primary open-angle glaucoma, the most common form of glaucoma, is a chronic, progressive disease often, though not always, accompanied by elevated intraocular pressure (IOP). In this disorder, retinal ganglion cell loss and excavation of the optic nerve head produce characteristic peripheral visual field deficits. Patients with normal-tension glaucoma present with typical visual field and optic nerve head changes, without a documented history of elevated IOP. A variety of secondary causes, such as pigment dispersion syndrome and ocular trauma, can result in glaucoma as well. Treatment of all forms of glaucoma consists of reducing IOP. With proper treatment, progression of this disease can often be delayed or prevented. Treatment options for glaucoma include medications, laser therapy and incisional surgery. Laser techniques for the reduction of IOP include argon laser trabeculoplasty and selective laser trabeculoplasty. Both techniques work by increasing outflow of aqueous humour through the trabecular meshwork. Surgical options for glaucoma treatment include trabeculectomy, glaucoma drainage tube implantation and ciliary body cyclodestruction. While each of these types of procedures is effective at lowering IOP, therapy usually begins with medications. Medications lower IOP either by reducing the production or by increasing the rate of outflow of aqueous humour within the eye. Currently, there are five major classes of drugs used for the treatment of glaucoma: (i) cholinergics (acetylcholine receptor agonists); (ii) adrenoceptor agonists; (iii) carbonic anhydrase inhibitors (CAIs); (iv) beta-adrenoceptor antagonists; and (v) prostaglandin analogues (PGAs). Treatment typically begins with the selection of an agent for IOP reduction. Although beta-adrenoceptor antagonists are still commonly used by many clinicians, the PGAs are playing an increasingly important role in the first-line therapy of glaucoma. Adjunctive agents, such as alpha-adrenoceptor agonists and CAIs are often effective at providing additional reduction in IOP for patients not controlled on monotherapy. As with any chronic disease, effective treatment depends on minimising the adverse effects of therapy and maximising patient compliance. The introduction of a variety of well tolerated and potent medications over the past few years now allows the clinician to choose a treatment regimen on an individual patient basis and thereby treat this disorder more effectively.

Cardiac arrhythmia monitoring during bronchial provocation test with methacholine.

STUDY OBJECTIVES: During a bronchial provocation test (BPT), the performance of maximal inspiratory-expiratory maneuvers, causing abrupt and marked shifts in intrathoracic pressure, may increase the risk of cardiac arrhythmias. Moreover, the inhalation of methacholine (MCh), a cholinergic agonist agent, could favor the development of unwelcome cardiovascular events, namely, cardiac arrhythmias. SUBJECTS AND METHODS: We studied the number and severity of cardiac arrhythmias by ECG-Holter monitoring before, during, and after BPTs with MCh challenge in a group of 46 consecutive nonselected subjects (28 men and 18 women) with clinical indications for BPT, without preexisting cardiovascular diseases, and not receiving arrhythmogenic drugs. The subjects performed a routine pulmonary function test (PFT), followed by BPT, during ECG-Holter monitoring. Determination of the serum potassium concentration, a baseline arterial blood gas analysis, and monitoring of oxyhemoglobin saturation also were performed. RESULTS: We found no significant increase in the number of supraventricular and ventricular arrhythmias during the performance of PFTs and of BPTs with MCh in the subjects, either with or without bronchial hyperresponsiveness (BHR). However, during the performance of BPTs, we observed a significant reduction in mean heart rate. CONCLUSIONS: Our results indicate that the performance of PFTs and BPTs with MCh does not increase the cardiac arrhythmogenic risk in subjects without cardiovascular diseases, as well as in those with BHR, suggesting that these tests are safe to perform in most subjects.

A pilot controlled clinical trial of ABT-418, a cholinergic agonist, in the treatment of adults with attention deficit hyperactivity disorder.

OBJECTIVE: Despite the increasing recognition of attention deficit hyperactivity disorder (ADHD) in adults, there is a paucity of controlled pharmacological trials. Recent reports have suggested the potential usefulness of cholinergic agents for ADHD. To this end, the authors completed a controlled study of ABT-418, a novel cholinergic activating agent, for the treatment of adults with ADHD. METHOD: This was a double-blind, placebo-controlled, randomized, crossover trial that compared a transdermal patch of ABT-418 (75 mg/day) to placebo in adults who met DSM-IV criteria for ADHD. There were two 3-week treatment periods separated by 1 week of washout. RESULTS: Of the 32 subjects enrolled in the study (88% were men; mean age = 40 years, SD = 9), 29 completed the study. At the endpoint of each active arm (last observation carried forward), a significantly higher proportion of subjects was considered improved while receiving ABT-418 than while receiving placebo (40% versus 13%). Similarly, at endpoint there was a significantly greater reduction in ADHD symptom checklist scores (28% versus 15%). Symptoms reflective of attention, and subjects with less severe ADHD, responded more robustly to ABT-418. Treatment with ABT-418 was relatively well tolerated; dizziness and nausea were the most frequently reported adverse effects. CONCLUSIONS: The results of this investigation indicate that ABT-418, a nicotinic analog, may be a potentially useful agent for the treatment of ADHD.