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Introduction: We have previously shown that Environmental Enrichment (EE), a multi-modal psychosocial intervention consisting of increased social interaction, novelty, and open spaces, improved disease presentation, anxiety, and immune-related disturbances in the rat model of endometriosis. However, there is a knowledge gap regarding the effects of EE interventions in patients with this painful, inflammatory chronic disease. Aim: To adapt and test the efficacy of an EE intervention on pelvic pain, mental health, perceived stress, quality of life, and systemic inflammation in endometriosis patients through a randomized clinical trial (RCT). Materials and methods: A multidisciplinary team with expertise in physiology, neuroscience, psychology, and women's health adapted and implemented a two-arm RCT comparing an EE intervention with a wait-list control group. Six EE modules administered on alternate weeks were provided to patients in the intervention (N = 29); controls received education only. Survey data and biospecimens were collected at baseline, end-of-study, and 3-months post-intervention to assess pain (Brief Pain Inventory, BPI), endometriosis-related quality of life-QoL (Endometriosis Health Profile-30, EHP30), anxiety (Generalized Anxiety Disorder 7, GAD7), depression (Patient Health Questionnaire for Depression 8, PHQ8), pain catastrophizing (Pain Catastrophizing Score, PCS), stress (Perceived Stress Scale-14, PSS14), and saliva cortisol levels (AM, PM). Results: Compared to the wait-list controls, participants in the EE intervention showed significantly decreased GAD-7 scores at the end of the intervention and 3-month follow-up. Depression, perceived stress, and QoL improved at the 3-month follow-up compared to baseline. While pain levels did not improve, they significantly correlated with anxiety, depression, QoL and pain catastrophizing scores. Conclusion: This pilot RCT demonstrated significant improvements in anxiety and depressive symptoms, QoL, and perceived stress, supporting enriched environments as an integrative psychosocial intervention to be used as adjuvant to the standard of care for endometriosis pain.
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Biomedical graduate students receive intensive training in their scientific area of interest yet need additional skills for successful scientific careers. Our aim was to promote team building, improve collaborations and enhance communication skills. An off-site yearly retreat was organized for the graduate students in our NIH-funded Research Initiative for Scientific Enhancement (RISE) graduate training program. Retreat themes were addressed through short presentations, case studies, live podcasts, webinars, focus groups, role-play, and breakout sessions with various team building exercises to practice communication skills and identify abilities, knowledge, values, and behaviors. Trainees gave short presentations and served as discussion leaders on topics related to the central theme. Expert guest speakers participated in discussion sessions with the trainees. Trainees evaluated the retreats at the end. A total of 48 trainees, 12 RISE Program faculty and staff, and 26 external speakers from industry, academia, media/journalism, the arts, psychology, and holistic medical fields participated over 9 years. The overall average benefit of the in-person retreats was rated 4.80 on a Likert scale of 1-5 by trainees. Trainees particularly enjoyed the informal interactions with program faculty, staff, and fellow trainees. They appreciated the opportunity to learn soft skills, such as interpersonal communication, conflict resolution, and leadership. Two additional retreats conducted virtually because of the COVID-19 pandemic were perceived as less beneficial. We conclude that off-site interactive retreats are a valuable tool for enhancing soft skills and a sense of team identity in a biomedical sciences graduate program, while covering important issues related to scientific careers.NEW & NOTEWORTHY Off-site interactive science-related retreats are a valuable tool for enhancing soft skills and sense of team identity in a biomedical sciences graduate program, while covering important issues related to pursuing a career in science. There are many perceived benefits, so we encourage other training programs to include a similar type of regular activity in students' training with the goal of improving trainee well-being and supporting their academic and research productivity.
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Docentes , Pandemias , Humanos , Educación de Postgrado en Medicina , Estudiantes , ComunicaciónRESUMEN
High-fat diet (HFD) is associated with gut microbiome dysfunction and mental disorders. However, the time-dependence as to when this occurs is unclear. We hypothesized that a short-term HFD causes colonic tissue integrity changes resulting in behavioral changes. Rats were fed HFD or low-fat diet (LFD) for a month and gut microbiome, colon, and behavior were evaluated. Behavioral despair was found in the HFD group. Although obesity was absent, the HFD group showed increased percent weight gain, epididymal fat tissue, and leptin expression. Moreover, the HFD group had increased colonic damage, decreased expression of the tight junction proteins, and higher lipopolysaccharides (LPS) in serum. Metagenomic analysis revealed that the HFD group had more Bacteroides and less S24-7 which correlated with the decreased claudin-5. Finally, HFD group showed an increase of microglia percent area, increased astrocytic projections, and decreased phospho-mTOR. In conclusion, HFD consumption in a short period is still sufficient to disrupt gut integrity resulting in LPS infiltration, alterations in the brain, and behavioral despair even in the absence of obesity.
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Introduction: We have previously shown that Environmental Enrichment (EE)-consisting of social support, novelty, and open spaces-decreased disease progression and anxiety in a rat model of endometriosis. We developed a novel EE intervention to be tested in a pilot randomized clinical trial (RCT) in patients with endometriosis, a painful, stressful disease. Objective: To translate and evaluate the feasibility and acceptability of an adapted EE intervention as an adjuvant to standard-of-care for endometriosis patients. Methods: Feasibility was assessed through recruitment, enrollment, and adherence rates. Acceptability was evaluated through a post-intervention survey and focus group discussion 3-months after the end of the intervention. Results: Of the 103 subjects recruited, 64 were randomized to the intervention group and 39 to the control group. At the start of the intervention, the study groups consisted of 29 (intervention) and 27 (control) subjects. Enrollment rates were 45.3% and 69.2%, and adherence rates were 41.4% and 100% for the intervention and control groups, respectively. Delays resulting from natural events (earthquakes, the COVID-19 pandemic) impacted enrollment and adherence rates. The most common reasons for missing an intervention were period pain (39.1%) and work-study (34.8%). There was high acceptability (>80%) of the intervention's logistics. The majority (82.4%) of subjects would continue participating in support groups regularly, and 95.7% would recommend the intervention to other patients. Conclusions: We showed that EE could be translated into an acceptable integrative multi-modal therapy perceived as valuable among participants who completed the intervention. High attrition/low adherence indicates that additional refinements would be needed to improve feasibility. Acceptability data indicate that EE has the potential to be integrated into the clinical management of patients with endometriosis and other inflammatory, painful disorders. Studies are ongoing to assess the efficacy of EE in improving pain symptoms, mental health, and quality of life (QoL).
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Endometriosis is a painful gynecological disease with no cure and limited therapeutic options. It has been hypothesized that epigenetic drugs can be used as a nonhormonal treatment for endometriosis. This study was conducted to study the efficacy of an inhibitor of the histone methyltransferase EZH2 using an established rat model of endometriosis. We hypothesized that treatment will block or reduce the number of endometriotic vesicles in this model. We conducted a preclinical drug study in female rats with experimental endometriosis (uterine tissue transplanted next to the intestinal mesentery) or control sham (sutures only). Rats with endometriosis or sham surgery received either treatment with EZH2 inhibitor (5 mg/kg or 10 mg/kg) or vehicle (0.1%, 67% DMSO) every other day during 4 weeks. After treatment completion, the number, area, volume, and weight of vesicles were evaluated. RT [2] Profiler Arrays for neuropathic and inflammation, epithelial to mesenchymal transition, inflammatory response, and autoimmunity pathways were used to examine gene expression changes in the vesicles that developed. Treatment with EZH2 inhibitor (10 mg/kg) suppressed the development of vesicles, by significantly decreasing the total vesicle number, area, volume, and weight. In addition, EZH2 inhibition significantly increased the expression of CACNA1B and FKBP1A genes, involved in pain and proliferation, respectively. EZH2 inhibition suppresses the growth of vesicles without apparent detrimental effects to other organs. Treatment with this epigenetic inhibitor leads to upregulation of a limited number of genes related to endometriosis-relevant pathways. In conclusion, these data support follow-up studies to evaluate its potential as a therapeutic approach for endometriosis.
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Endometriosis/metabolismo , Endometrio/metabolismo , Proteína Potenciadora del Homólogo Zeste 2/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , Animales , Modelos Animales de Enfermedad , Endometriosis/genética , Endometriosis/patología , Endometrio/efectos de los fármacos , Endometrio/patología , Transición Epitelial-Mesenquimal/efectos de los fármacos , Transición Epitelial-Mesenquimal/fisiología , Femenino , Indazoles/farmacología , Piridonas/farmacología , Ratas , Ratas Sprague-DawleyRESUMEN
PURPOSE: We have previously shown that stress prior to induction worsens clinical presentation and inflammatory parameters in a rat model of endometriosis. This study was designed to examine whether stress during the development of endometriosis can affect the growth of endometriotic implants through nerve growth and immune alterations. METHODS: Endometriosis was surgically induced in female Sprague-Dawley rats by suturing uterine horn implants onto the small intestine mesentery. Two weeks later, one group of rats (endo-stress) was subjected to a 10-day swim stress protocol. Controls had no stress (endo-no stress) or sutures only and stress (sham-stress). On day 60, all rats were killed and examined for the presence of endometriotic vesicles. The size of each vesicle was measured. The uterus and colon were removed and assessed for damage, cell infiltration, and expression of nerve growth factor (NGF), its receptors (p75 and Tropomyosin receptor kinase A (Trk-A)/pTrk-A), and calcitonin gene-related peptide, a sensory fiber marker. A differential analysis of peritoneal fluid white blood cell count was performed. RESULTS: Stress significantly increased endometriotic vesicle size but not colonic damage and increased infiltration of mast cells. Significantly increased expression of NGF and its receptors was found in the uterus of animals with endometriosis receiving stress. CONCLUSIONS: Stress stimulates the development of ectopic endometrial vesicles in an animal model of endometriosis and increases inflammatory cell recruitment to the peritoneum. In addition, stress promotes nerve fiber growth in the uterus.
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Endometriosis/metabolismo , Factor de Crecimiento Nervioso/metabolismo , Neurogénesis/fisiología , Receptor trkA/metabolismo , Receptores de Factor de Crecimiento Nervioso/metabolismo , Estrés Fisiológico/fisiología , Estrés Psicológico/metabolismo , Animales , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Endometriosis/patología , Femenino , Proteínas del Tejido Nervioso , Fosforilación , Ratas , Ratas Sprague-Dawley , Receptores de Factores de Crecimiento , Estrés Psicológico/patologíaRESUMEN
Treatments for endometriosis include pharmacological or surgical procedures that produce significant side effects. We aimed to determine how environmental enrichment (EE) could impact the progression of endometriosis using the autotransplantation rat model. Female rats were exposed to EE (endo-EE: toys and nesting materials, 4 rats per cage, larger area enclosure) or no enrichment (endo-NE: 2 rats per cage) starting on postnatal day 21. After 8 weeks, sham surgery or surgical endometriosis was induced by suturing uterine horn tissue next to the intestinal mesentery, then allowed to progress for 60 days during which EE or NE continued. At the time of killing, we measured anxiety behaviors, collected endometriotic vesicles and uterus, and processed for quantitative real-time polymerase chain reaction for corticotropin-releasing hormone (CRH), urocortin-1, CRH receptors type 1 and type 2, and glucocorticoid receptor (GR). Endometriosis did not affect anxiety-like behaviors, yet rats in enriched conditions showed lower basal anxiety behaviors than the nonenriched group. Importantly, the endo-EE group showed a 28% reduction in the number of endometriosis vesicles and the vesicles were significantly smaller compared to the endo-NE group. Endometriosis increased CRH and GR only in the vesicles of endo-NE, and this increase was dampened in the endo-EE. However, urocortin 1 was increased in the vesicles of the endo-EE group, suggesting different pathways of activation of CRH receptors in this group. Our results suggest that the use of multimodal complementary therapies that reduce stress in endometriosis could be an effective and safe treatment alternative, with minimal side effects.
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Conducta Animal/fisiología , Endometriosis/terapia , Ambiente , Vivienda para Animales , Útero/metabolismo , Animales , Ansiedad/metabolismo , Ansiedad/psicología , Hormona Liberadora de Corticotropina/metabolismo , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Endometriosis/metabolismo , Endometriosis/psicología , Femenino , Ratas , Ratas Sprague-Dawley , Receptores de Hormona Liberadora de Corticotropina/metabolismo , Receptores de Glucocorticoides/metabolismo , Estrés Psicológico/metabolismo , Estrés Psicológico/psicología , Urocortinas/metabolismoRESUMEN
AIM: To determine serum vitamin D levels and colonic vitamin D receptor (VDR) expression in inflammatory bowel disease (IBD) and non-IBD patients and correlate these with histopathology. METHODS: Puerto Rican IBD (n = 10) and non-IBD (n = 10) patients ≥ 21 years old scheduled for colonoscopy were recruited. Each patient completed a questionnaire and provided a serum sample and a colonic biopsy of normal-appearing mucosa. For IBD patients, an additional biopsy was collected from visually diseased mucosa. Serum vitamin D levels were measured by ultra-performance liquid chromatography and mass spectrometry. Hematoxylin and eosin stained tissue sections from colonic biopsies were classified histologically as normal or colitis (active/inactive), and scored for the degree of inflammation present (0-3, inactive/absent to severe). Tissue sections from colonic biopsies were also stained by immunohistochemistry for VDR, for which representative diagnostic areas were photographed and scored for staining intensity using a 4-point scale. RESULTS: The IBD cohort was significantly younger (40.40 ± 5.27, P < 0.05) than the non-IBD cohort (56.70 ± 1.64) with a higher prevalence of vitamin D deficiency (40% vs 20%, respectively) and insufficiency (70% vs 50%, respectively). Histologic inflammation was significantly higher in visually diseased mucosa from IBD patients (1.95 ± 0.25) than in normal-appearing mucosa from control patients (0.25 ± 0.08, P < 0.01) and from IBD patients (0.65 ± 0.36, P < 0.05) and correlated inversely with VDR expression in visually diseased colonic tissue from IBD patients (r = -0.44, P < 0.05) and from IBD patients with Crohn's disease (r = -0.69, P < 0.05), but not in normal-appearing colonic tissue from control patients or IBD patients. Control and IBD patient serum vitamin D levels correlated positively with VDR expression in normal colon from control and IBD patients (r = 0.38, P < 0.05) and with patient age (r = 0.54, P < 0.01). CONCLUSION: Levels of serum vitamin D correlate positively with colonic VDR expression in visually normal mucosa whereas inflammation correlates negatively with colonic VDR expression in visually diseased mucosa in Puerto Rican patients.
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Colitis Ulcerosa/sangre , Colon/química , Enfermedad de Crohn/sangre , Mucosa Intestinal/química , Receptores de Calcitriol/análisis , Deficiencia de Vitamina D/sangre , Vitamina D/análogos & derivados , Adulto , Anciano , Biomarcadores/sangre , Biopsia , Estudios de Casos y Controles , Cromatografía Liquida , Colitis Ulcerosa/diagnóstico , Colitis Ulcerosa/epidemiología , Colon/patología , Enfermedad de Crohn/diagnóstico , Enfermedad de Crohn/epidemiología , Femenino , Humanos , Inmunohistoquímica , Mucosa Intestinal/patología , Masculino , Espectrometría de Masas , Persona de Mediana Edad , Prevalencia , Puerto Rico/epidemiología , Vitamina D/sangre , Deficiencia de Vitamina D/diagnóstico , Deficiencia de Vitamina D/epidemiología , Adulto JovenRESUMEN
The use and validity of the Graduate Record Examination General Test (GRE) to predict the success of graduate school applicants is heavily debated, especially for its possible impact on the selection of underrepresented minorities into science, technology, engineering, and math fields. To better identify candidates who would succeed in our program with less reliance on the GRE and grade point average (GPA), we developed and tested a composite score (CS) that incorporates additional measurable predictors of success to evaluate incoming applicants. Uniform numerical values were assigned to GPA, GRE, research experience, advanced course work or degrees, presentations, and publications. We compared the CS of our students with their achievement of program goals and graduate school outcomes. The average CS was significantly higher in those students completing the graduate program versus dropouts (p < 0.002) and correlated with success in competing for fellowships and a shorter time to thesis defense. In contrast, these outcomes were not predicted by GPA, science GPA, or GRE. Recent implementation of an impromptu writing assessment during the interview suggests the CS can be improved further. We conclude that the CS provides a broader quantitative measure that better predicts success of students in our program and allows improved evaluation and selection of the most promising candidates.
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Investigación Biomédica/educación , Evaluación Educacional , Estudiantes , Adulto , Demografía , Becas , Femenino , Humanos , Masculino , Persona de Mediana Edad , Puerto Rico , Criterios de Admisión Escolar , Adulto JovenRESUMEN
AIM: To determine the expression of neurokinin-1 receptor (NK-1R), phosphorylated epidermal growth factor receptor (pEGFR), cyclooxygenase-2 (Cox-2), and vitamin D receptor (VDR) in normal, inflammatory bowel disease (IBD), and colorectal neoplasia tissues from Puerto Ricans. METHODS: Tissues from patients with IBD, colitis-associated colorectal cancer (CAC), sporadic dysplasia, and sporadic colorectal cancer (CRC), as well as normal controls, were identified at several centers in Puerto Rico. Archival formalin-fixed, paraffin-embedded tissues were de-identified and processed by immunohistochemistry for NK-1R, pEGFR, Cox-2, and VDR. Pictures of representative areas of each tissues diagnosis were taken and scored by three observers using a 4-point scale that assessed intensity of staining. Tissues with CAC were further analyzed by photographing representative areas of IBD and the different grades of dysplasia, in addition to the areas of cancer, within each tissue. Differences in the average age between the five patient groups were assessed with one-way analysis of variance and Tukey-Kramer multiple comparisons test. The mean scores for normal tissues and tissues with IBD, dysplasia, CRC, and CAC were calculated and statistically compared using one-way analysis of variance and Dunnett's multiple comparisons test. Correlations between protein expression patterns were analyzed with the Pearson's product-moment correlation coefficient. Data are presented as mean ± SE. RESULTS: On average, patients with IBD were younger (34.60 ± 5.81) than normal (63.20 ± 6.13, P < 0.01), sporadic dysplasia (68.80 ± 4.42, P < 0.01), sporadic cancer (74.80 ± 4.91, P < 0.001), and CAC (57.50 ± 5.11, P < 0.05) patients. NK-1R in cancer tissue (sporadic CRC, 1.73 ± 0.34; CAC, 1.57 ± 0.53) and sporadic dysplasia (2.00 ± 0.45) were higher than in normal tissues (0.73 ± 0.19). pEGFR was significantly increased in sporadic CRC (1.53 ± 0.43) and CAC (2.25 ± 0.47) when compared to normal tissue (0.07 ± 0.25, P < 0.05, P < 0.001, respectively). Cox-2 was significantly increased in sporadic colorectal cancer (2.20 ± 0.23 vs 0.80 ± 0.37 for normal tissues, P < 0.05). In comparison to normal (2.80 ± 0.13) and CAC (2.50 ± 0.33) tissues, VDR was significantly decreased in sporadic dysplasia (0.00 ± 0.00, P < 0.001 vs normal, P < 0.001 vs CAC) and sporadic CRC (0.47 ± 0.23, P < 0.001 vs normal, P < 0.001 vs CAC). VDR levels negatively correlated with NK-1R (r = -0.48) and pEGFR (r = -0.56) in normal, IBD, sporadic dysplasia and sporadic CRC tissue, but not in CAC. CONCLUSION: Immunohistochemical NK-1R and pEGFR positivity with VDR negativity can be used to identify areas of sporadic colorectal neoplasia. VDR immunoreactivity can distinguish CAC from sporadic cancer.
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Biomarcadores de Tumor/análisis , Neoplasias Colorrectales/química , Receptores ErbB/análisis , Inmunohistoquímica , Enfermedades Inflamatorias del Intestino/metabolismo , Receptores de Calcitriol/análisis , Receptores de Neuroquinina-1/análisis , Adulto , Anciano , Biopsia , Estudios de Casos y Controles , Neoplasias Colorrectales/etnología , Neoplasias Colorrectales/patología , Ciclooxigenasa 2/análisis , Femenino , Hispánicos o Latinos , Humanos , Enfermedades Inflamatorias del Intestino/etnología , Enfermedades Inflamatorias del Intestino/patología , Masculino , Persona de Mediana Edad , Clasificación del Tumor , Fosforilación , Puerto Rico/epidemiologíaRESUMEN
Barriers persist in the development and delivery of effective cancer therapies to under-represented minority populations. In Puerto Rico, cancer is the second leading cause of death, yet cancer research awareness and training opportunities remain somewhat limited on the island. These limitations hinder progress toward decreasing the cancer health disparities that exist within the Puerto Rican population. The predominantly Hispanic population of Puerto Rico is the focus of a partnership between the Ponce Health Sciences University-Medical School and Ponce Research Institute (PHSU) in Ponce, Puerto Rico and the H. Lee Moffitt Cancer Center in Tampa, Florida. The Partnership goals are to reduce these barriers through an integrated, multipronged approach of training and education alongside outreach and research components. This report describes the approaches, successes and challenges of enhancing clinical cancer research capacity on the island and the unique challenges of a partnership between two institutes physically separated by long distances. Once fully developed this model may be exportable to other Latin American countries where the need is even greater.
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Investigación Biomédica/estadística & datos numéricos , Educación Médica/métodos , Hispánicos o Latinos , Oncología Médica/educación , Neoplasias/etnología , Humanos , Neoplasias/prevención & control , Puerto Rico/etnología , Estados Unidos/epidemiologíaRESUMEN
UNLABELLED: Little is known about the effects of the pro-inflammatory hormone Angiotensin II (Ang II) in inflammatory bowel disease. The aim of this study was to evaluate the effect of valsartan (Diovan), an Ang II receptor antagonist, in two models of colitis. METHODS: Colitis was induced in Sprague-Dawley rats by administration of trinitrobenzene sulfonic acid (TNBS; 30 mg in 50% ETOH i.c.) or 5% Dextran Sulphate Sodium (DSS) in drinking water ad libitum for 5 days. Valsartan was administered orally in drinking water (160 mg/L) during thirty days prior to the induction of the colitis, and for 5 days after. All animals were evaluated for weight change, diarrhea, myeloperoxidase activity, macroscopic and microscopic damage. Cytokine levels in the colon were measured by ELISA, real-time RT-PCR and immunohistochemistry. RESULTS: In the TNBS model, valsartan reduced the macroscopic damage score, significantly decreased the microscopic damage (p<0.01), and accelerated weight gain after colitis. In the DSS-colitis model, valsartan-treated animals had less diarrhea and microscopic damage. Valsartan reduced the protein levels of TGFbeta (p<0.05), and IL-18 in the TNBS model, and led to over expression of IL-10 mRNA in the DSS model. CONCLUSION: These data demonstrate a possible anti-inflammatory effect for valsartan in colitis.
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Antagonistas de Receptores de Angiotensina , Antihipertensivos/uso terapéutico , Colitis/tratamiento farmacológico , Enfermedades Inflamatorias del Intestino/tratamiento farmacológico , Tetrazoles/uso terapéutico , Valina/análogos & derivados , Animales , Colitis/inducido químicamente , Citocinas/metabolismo , Modelos Animales de Enfermedad , Humanos , Inmunohistoquímica , Enfermedades Inflamatorias del Intestino/inmunología , Masculino , Ratas , Ratas Sprague-Dawley , Valina/uso terapéutico , ValsartánRESUMEN
Endometriosis commonly presents with symptoms that mimic chronic gastrointestinal disorders. The authors used the autotransplantion model of endometriosis in rats to investigate the possible underlying mechanisms. After the rats were killed, the presence of endometriotic vesicles, colonic inflammation, and white blood cell (WBC) numbers in the peritoneal fluid was determined. Sections of colon and of jejunum were collected for measurement of myeloperoxidase (MPO) activity and bacterial counts, and isometric recording in response to acetylcholine was measured in segments of longitudinal and circular smooth muscle. Experimental animals had significantly more colonic damage, MPO activity, and WBC numbers than controls did. There was no significant difference in the total bacterial load; however, experimental animals demonstrated an increased tension in the longitudinal muscle, which correlated with WBC numbers and colonic damage. In summary, this study presents evidence for a significant effect of peritoneal endometriosis on colonic function and integrity, which may help explain the gastrointestinal symptoms associated with this disease.
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Endometritis/fisiopatología , Motilidad Gastrointestinal/fisiología , Lactobacillus/aislamiento & purificación , Animales , Colitis/enzimología , Colitis/inmunología , Colitis/microbiología , Recuento de Colonia Microbiana , Modelos Animales de Enfermedad , Endometritis/enzimología , Endometritis/microbiología , Femenino , Técnicas In Vitro , Contracción Muscular/fisiología , Peroxidasa/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
Inflammatory bowel disease (IBD) is a gastrointestinal disorder of unknown etiology or cure. One complication of IBD is an increased risk for development of colon cancer. The aims of this study were to use a previously established rat model of colitis to develop a new model of colitis-associated colon cancer and ascertain the involvement of three cancer-related genes: K-ras, adenomatous polyposis coli (APC), and p53. Four groups of rats were used: reactivated 1,2-dimethylhydrazine [DMH; trinitrobenzene sulfonic acid (TNBS) was used to induce colitis followed by a weekly s.c. dose of DMH], prolonged reactivation (inflammation was induced with TNBS, then maintained twice a week), saline-DMH (animals received saline instead of TNBS followed by a weekly dose of DMH), and normal (received no treatment). Animals were sacrificed at 5, 10, or 15 weeks, and colon samples were taken for pathologic analysis and gene mutation detection. No dysplasia was found in the normal group. The highest incidences of dysplasia were as follows: prolonged reactivation group at 5 weeks (60%), reactivated DMH group at 10 weeks (83%), and saline-DMH group at 15 weeks (67%). Carcinoma was found in both the prolonged reactivation and saline-DMH groups. No mutations were found in the K-ras oncogene; however 62% of the APC samples (exon 15 at nucleotide 2778) and 76% of p53 (exon 6 at nucleotide 1327) showed substitutions. The prolonged reactivation group may be considered a new model of colitis-associated colon cancer, offering the potential to study cancer prevention strategies for patients with IBD.
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Neoplasias del Colon/inmunología , Neoplasias del Colon/patología , Inflamación/patología , 1,2-Dimetilhidrazina/farmacología , Proteína de la Poliposis Adenomatosa del Colon/metabolismo , Animales , Carcinoma/genética , Carcinoma/metabolismo , Enfermedad Crónica , Colitis/inducido químicamente , Colitis/metabolismo , Colitis/patología , Neoplasias del Colon/genética , Exones , Enfermedades Inflamatorias del Intestino/metabolismo , Enfermedades Inflamatorias del Intestino/patología , Mutación , Ratas , Factores de Tiempo , Ácido Trinitrobencenosulfónico/farmacología , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
BACKGROUND: Inflammatory bowel disease (IBD) is a disease of unknown etiology characterized by acute and chronic relapsing inflammation. The most suitable animal model for studying this disease is still under debate. Microarray transcript profiling has the potential to illuminate the molecular processes that are involved in both the human condition and animal models. AIM: To identify differentially expressed genes in the 2,4,6-trinitrobenzene sulfonic acid (TNBS) model of experimental colitis and compare gene expression profiles with that reported in patients. METHODS: Colitis was induced by TNBS administration (30 mg in 50% ethanol) in female Sprague-Dawley rats. Controls received the vehicle. Seventy-two hours later, the animals were killed, the colons were removed and scored for damage, and total RNA was isolated. Gene expression levels were analyzed after hybridizing experimental and control cDNA to PIQOR Toxicology Rat Microarrays containing 1,252 genes. Genes with 2-fold or more higher or 0.5-fold or less lower expression levels were selected as significantly differentially expressed. Results were validated using real-time reverse-transcription polymerase chain reaction (RT-PCR). RESULTS: We observed increased expression of genes that have previously been shown to be up-regulated in IBD patients, including chemokines/cytokines, extracellular matrix/remodeling genes, transcription factors and tumor necrosis factor family members. Using real-time RT-PCR, we validated 9 of 10 critical genes identified by DNA microarray. Fibulin 2 and lysyl oxidase are among some of the novel genes not previously associated with IBD that could potentially be related to the pathogenesis of this condition. CONCLUSION: We provide evidence supporting the TNBS colitis model as an appropriate tool to study the pathology of IBD and identify molecular targets with clinical relevance.
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Colitis/genética , Perfilación de la Expresión Génica/métodos , Mediadores de Inflamación/metabolismo , Animales , Biomarcadores/metabolismo , Colitis/inducido químicamente , Colitis/metabolismo , Modelos Animales de Enfermedad , Procesamiento Automatizado de Datos , Femenino , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Ácido TrinitrobencenosulfónicoRESUMEN
It is known that the muscularis mucosae and mucosa are not pharmacologically homogeneous throughout the rat colon. The aim of this study was to simultaneously characterize all three neurokinin (NK) receptors in the muscularis mucosae and mucosa along the length of the rat colon. Strips of proximal, mid, and distal colonic muscularis mucosae were prepared for isometric recording or sheets of muscle-free mucosa were mounted in Ussing chambers for measurement of short-circuit current. In both muscularis mucosae and mucosa the greatest responses to substance P were found in the proximal region. Use of selective agonists revealed the presence of all three NK receptors in both structures, however, selective antagonism suggests that only NK2 receptors in the muscularis mucosae and NK1 receptors in the mucosa are physiologically relevant. In conclusion, substance P-induced responses in the rat colon are region-specific and not mediated by a single NK receptor subtype common to both structures.
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Acetilcolina/farmacología , Colon/efectos de los fármacos , Motilidad Gastrointestinal/efectos de los fármacos , Mucosa Intestinal/efectos de los fármacos , Receptores de Taquicininas/metabolismo , Sustancia P/farmacología , Animales , Colon/patología , Modelos Animales de Enfermedad , Femenino , Motilidad Gastrointestinal/fisiología , Mucosa Intestinal/patología , Masculino , Músculo Liso/efectos de los fármacos , Músculo Liso/metabolismo , Ratas , Ratas Sprague-Dawley , Receptores de Neuroquinina-1/efectos de los fármacos , Receptores de Neuroquinina-1/metabolismo , Receptores de Neuroquinina-2/efectos de los fármacos , Receptores de Neuroquinina-2/metabolismo , Receptores de Taquicininas/efectos de los fármacos , Sensibilidad y Especificidad , Técnicas de Cultivo de TejidosRESUMEN
Endometriosis is commonly associated with symptoms similar to those of gastrointestinal diseases, such as inflammatory bowel disease (IBD), leading to erroneous diagnosis and inappropriate management. The role of tumor necrosis factor alpha (TNF) in IBD is well established, but its role in endometriosis--also characterized by the activation of inflammatory mechanisms--is still under study. Furthermore, little is known about the involvement of TNF receptors. Intestinal endometriosis was surgically induced in female Sprague-Dawley rats (n = 10). Control rats (n = 10) received sutures with no implants. Samples of tissue and fluids were collected 60 days after surgery. Endometriotic implants were classified in grades, and the gastrointestinal tract was examined for damage. A significant increase was observed in protein levels of TNF and soluble TNFRSF1B in the peritoneal fluid of experimental rats compared to controls. Expression of Tnf mRNA was significantly increased both in peritoneal leukocytes and in intestinal segments associated with implants in experimental animals. Bioactivity of TNF in tissues was confirmed by overexpression of Icam1, Sele, Vegfa, Flt1 and Kdr. Gene expression of Tnfrsf1a and Tnfrsf1b was downregulated in colon and small intestine of experimental animals, possibly as a mechanism of protection against TNF cytotoxicity. Significant overexpression of genes encoding TNF receptor-associated factors that have been linked to activation of antiapoptotic pathways also was observed. Overexpression of TNF and target genes, underexpression of TNF-receptor genes, and increased shedding of TNFRSF1B in this animal model provide further evidence for involvement of the TNF system in the pathogenesis of endometriosis.
Asunto(s)
Endometriosis/genética , Enfermedades Intestinales/genética , Receptores Tipo II del Factor de Necrosis Tumoral/genética , Receptores Tipo II del Factor de Necrosis Tumoral/metabolismo , Receptores Tipo I de Factores de Necrosis Tumoral/genética , Animales , Modelos Animales de Enfermedad , Regulación hacia Abajo , Selectina E/genética , Endometriosis/metabolismo , Endometriosis/patología , Femenino , Regulación de la Expresión Génica , Molécula 1 de Adhesión Intercelular , Enfermedades Intestinales/metabolismo , Enfermedades Intestinales/patología , Recuento de Leucocitos , Peritoneo/citología , Peritoneo/patología , Ratas , Ratas Sprague-Dawley , Receptores Tipo I de Factores de Necrosis Tumoral/metabolismo , Solubilidad , Factor 1 Asociado a Receptor de TNF/genética , Factor 2 Asociado a Receptor de TNF/genética , Factor de Necrosis Tumoral alfa/metabolismo , Factor A de Crecimiento Endotelial Vascular/genética , Receptor 1 de Factores de Crecimiento Endotelial Vascular/genética , Receptor 2 de Factores de Crecimiento Endotelial Vascular/genéticaRESUMEN
BACKGROUND: Previous epidemiological studies have suggested that the incidence of inflammatory bowel disease (IBD) is lower in Latin American populations. The aim of this study was to estimate the incidence of IBD in Puerto Rico, a predominantly Hispanic population. METHODS: A nonconcurrent prospective study was conducted in collaboration with private gastroenterologists in southwest Puerto Rico. Basic medical history and demographics were extracted from the medical records of patients for which a new diagnosis of IBD (Crohn's disease, CD; ulcerative colitis, UC; or nonspecified IBD) was made during each of the years 1996-2000. RESULTS: A total of 202 eligible cases of IBD were identified (95 male, 107 female). Forty-eight patients were diagnosed with CD, 102 with UC, and 52 with nonspecified IBD. The total incidence of IBD increased significantly between 1996 and 2000 (3.07/100,000 to 7.74/100,000; p < 0.001), being significantly higher for CD (four-fold increase, p < 0.01) and nonspecified IBD (fourfold increase, p < 0.005), but not UC (1.7-fold increase). The prevalence of CD was higher in males with an earlier age of onset (p < 0.05). CONCLUSIONS: This study demonstrates that the incidence of IBD within the Puerto Rican population is increasing and may be higher than previously reported for other Latin American populations.
Asunto(s)
Colitis Ulcerosa/diagnóstico , Colitis Ulcerosa/epidemiología , Enfermedad de Crohn/diagnóstico , Enfermedad de Crohn/epidemiología , Adolescente , Adulto , Distribución por Edad , Edad de Inicio , Anciano , Anciano de 80 o más Años , Análisis de Varianza , Estudios de Cohortes , Femenino , Humanos , Enfermedades Inflamatorias del Intestino/diagnóstico , Enfermedades Inflamatorias del Intestino/epidemiología , Masculino , Persona de Mediana Edad , Prevalencia , Probabilidad , Pronóstico , Estudios Prospectivos , Puerto Rico/epidemiología , Medición de Riesgo , Índice de Severidad de la Enfermedad , Distribución por SexoRESUMEN
BACKGROUND: The involvement of enteropathogenic microorganisms in the pathogenesis of inflammatory bowel disease (IBD) and their importance in the different phases of inflammation are still unknown. AIM: To quantify the aerobic bacterial load in models of acute and chronic 'reactivated' colitis, and correlate this with damage. METHODS: Acute colitis was induced in rats by administration of trinitrobenzene sulfonic acid (TNBS) or dextran sulfate sodium (DSS). Colitis was 'reactivated' 6 weeks later by intravenous administration of TNBS. The distal colon was removed and scored macroscopically before inoculating samples. RESULTS: Bacterial load in rats with acute colitis (72 h) and chronic 'reactivated' colitis or their controls was significantly higher than untreated (p < 0.05); however, there were significantly more bacteria in acute colitis than in chronic 'reactivated' or their controls (p < 0.05). Both acute and chronic 'reactivated' colitis had significantly higher damage scores than untreated animals (p < 0.05). Bacterial load and damage score were significantly correlated only with acute colitis. CONCLUSIONS: The role of enteric microflora in the pathogenesis of IBD is greater during the acute phase of colitis. The correlation between bacterial load and tissue damage suggests that damage contributes to bacterial multiplication and exacerbation of colitis. Normal colonic flora may contribute to the relapse of the disease.