The influence of interstitial cells of Cajal density in the outcomes of pyeloplasty in adults: A prospective analysis.

PURPOSE: To evaluate if the density of interstitial cells of Cajal (ICC) in the ureteropelvic junction (UPJ) influences the outcomes of pyeloplasty in adults. METHODS: Twenty-three patients with the diagnosis of ureteropelvic junction obstruction (UPJO) that underwent laparoscopic dismembered pyeloplasty were included. ICC density was measured using immunohistochemistry reaction for c-KIT expression in the resected UPJ segment. Pyeloplasty outcome was evaluated by patient self-report pain, urinary outflow using DTPA renogram and hydronephrosis assessment using ultrasound (US) at 12 months of follow-up. A logistic regression analysis was performed to assess the association of pyeloplasty outcomes and ICC density. RESULTS: Low, moderate, and high ICC density were present in 17.4%, 30.4%, and 52.2% of the patients, respectively. Complete pain resolution was observed in 100%, 85.7%, and 75% of patients with low, moderate and high ICC density, respectively (p = 0.791). DTPA renogram improved in 75%, 85.7%, and 91.7% of patients with low, moderate and high ICC density, respectively (p = 0.739). Hydronephrosis improved in 25%, 85.7%, and 91.7% of patients with low, moderate and high ICC density, respectively (p = 0.032). CONCLUSIONS: Patients with high ICC density have a significant amelioration of hydronephrosis after pyeloplasty. However, ICC density is not associated with functional outcomes.

Luteolin ameliorates loperamide-induced functional constipation in mice

Functional constipation (FC) is one of the most common gastrointestinal disorders characterized by hard stools and infrequent bowel movements, which is associated with dysfunction of the enteric nervous system and intestinal motility. Luteolin, a naturally occurring flavone, was reported to possess potential pharmacological activities on intestinal inflammation and nerve injury. This study aimed to explore the role of luteolin and its functional mechanism in loperamide-induced FC mice. Our results showed that luteolin treatment reversed the reduction in defecation frequency, fecal water content, and intestinal transit ratio, and the elevation in transit time of FC models. Consistently, luteolin increased the thickness of the muscular layer and lessened colonic histopathological injury induced by loperamide. Furthermore, we revealed that luteolin treatment increased the expression of neuronal protein HuC/D and the levels of intestinal motility-related biomarkers, including substance P (SP), vasoactive intestinal polypeptide (VIP), and acetylcholine (ACh), as well as interstitial cells of Cajal (ICC) biomarker KIT proto-oncogene, receptor tyrosine kinase (C-Kit), and anoctamin-1 (ANO1), implying that luteolin mediated enhancement of colonic function and contributed to the anti-intestinal dysmotility against loperamide-induced FC. Additionally, luteolin decreased the upregulation of aquaporin (AQP)-3, AQP-4, and AQP-8 in the colon of FC mice. In summary, our data showed that luteolin might be an attractive option for developing FC-relieving medications.

Gist sincrónico con adenomioma gástrico: Reporte de caso y revisión de la literatura / Synchronous gist with gastric adenomyoma: Case report and literature review

Introducción: La colisión de dos tumores de diferente estirpe celular en un mismo órgano es infrecuente; a pesar de las asociaciones descritas en la literatura, el hallazgo de GIST con adenomioma en sincronismo, llama aún más la atención debido a sus distintos orígenes celulares. Reporte de caso: Presentamos el caso de una paciente mujer de 57 años de edad, quien es sometida a cirugía de resección doble en cuña, y distintos exámenes incluido el anátomo-patológico. Conclusión: Se demuestra la presencia de tumores sincrónicos, GIST gástrico y adenomioma gástrico, a pesar de la infrecuencia de este hallazgo.

Background:The collision of two tumors of different cell lines in the same organ is infrequent; even though, the associations described in the literature, the finding of synchronous GISTwith adenomyoma draws even more attention due to its different cellular origins. We present the case of a 57-year-Case report:old female patient who underwent double wedge resection surgery and various examinations, including pathology. Conclusion:The presence of synchronous tumors, gastric GIST and gastric adenomyoma is demonstrated,despite the infrequency of this finding.

Neural crest derivatives and neuroendocrine cells in the gut of anencephalic and fetuses without congenital defects.

BACKGROUND: The enteric nervous system (ENS), a component of the peripheral nervous system in the intestinal walls, regulates motility, secretion, absorption, and blood flow. Neural crest (NC) migration, fundamental for ENS development, may be altered by central nervous system development alterations, such as neural tube defects (NTD). Intestinal innervation anomalies have been correlated to NTD. We aim to describe the ENS on a fetus with NTD and fetuses without congenital defects (FWCD). CASES: Two male and four female FWCD, 18-20 weeks-gestation (WG), and a 25 WG female anencephalic fetus. Samples from the pancreatoduodenal groove, jejunum, cecum, rectum, and appendix were analyzed by immunohistochemistry. Nervous plexuses were marked with Neuron-specific enolase and S-100; enteric glial cells with CD56; neuroendocrine cells with chromogranin and synaptophysin, and interstitial cells of Cajal (ICC) with CD117. RESULTS AND CONCLUSION: The anencephalic fetus presented a rudimentary brainstem with a cerebellum. Partial frontal, temporal, and occipital bones were found. A large atrial septal defect, an enlarged kidney with a duplex collecting system and a single adrenal gland were found. NSE, S100, and CD56, showed the presence of the myenteric and submucous plexuses of the ENS; scarce interplexus reactivity may indicate inadequate development. Pancreatic and gut neuroendocrine cells, identified with chromogranin and CD56, showed that the enteroendocrine system is present. Findings on FWCD using these markers are consistent with literature descriptions. Vagal NC migration appears to be unaffected despite the presence of anencephaly, although maturation of the ENS may be altered.

Esophagogastric junction outflow obstruction: Characterization of a new entity? Clinical, manometric, and neuroimmunological description.

OBJECTIVE: To determine the differences between clinical, manometric, and neuroimmunological profile of esophagogastric junction outflow obstruction (EGJOO) and achalasia patients. METHODS: Seven EGJOO and 27 achalasia patients were enrolled in a blind cross-sectional study. Peripheral blood (PB) of 10 healthy donors and 10 lower esophageal sphincter (LES) muscle biopsies from organ transplant donors were included as controls. The presence of ganglion cells, cells of Cajal, Th22/Th7/Th2/Th1/Tregs/Bregs/pDCregs in tissue, and PB was assessed by immunohistochemistry and flow cytometry. Serum concentration of IL-22/IL-17A/IL-17F/IL-4/IFN-γ/IL-1ß/IL-6/IL-23/IL-33/TNF-α/IL-10 was determined using bioplex plates. ANAs and antineuronal antibodies were evaluated by immunofluorescence and Western blot. KEY RESULTS: EGJOO and achalasia patients had lower ganglion cells and cells of Cajal percentage vs. controls, while fibrosis was present only in achalasia patients. EGJOO and controls had lower cell percentage of Th22/Th17/Th2 vs. achalasia. EGJOO tissue had lower Th1/Treg cell number vs. achalasia, but higher levels vs. control group. Bregs and pDCregs percentage was higher in EGJOO vs. control group. Percentage of PB subpopulations in EGJOO was not significantly different from control group. Serum cytokine levels were higher for IL-1ß/IL-6/TNF-α, while IL-17A levels were lower in EGJOO vs. achalasia and control group. EGJOO group was negative for ANAs, while in achalasia group, 54% were positive. GAD65 and PNMa/Ta2 antibodies were present in achalasia, whereas Yo and recoverin were positive in EGJOO group. CONCLUSIONS AND INFERENCES: Although EGJOO shares some clinical characteristics with achalasia, the neuroimmunological profile is completely different, suggesting that EGJOO might be a different entity.

Anti- and pro-oxidant effects of quercetin stabilized by microencapsulation on interstitial cells of Cajal, nitrergic neurons and M2-like macrophages in the jejunum of diabetic rats.

Given the well-known antioxidant and neuroprotective properties of quercetin, the aim of this work was to evaluate the effects of quercetin stabilized by microencapsulation at two doses (10 mg kg-1 and 100 mg kg-1) on the oxidative/antioxidant status, number and morphological features of ICC, nitrergic neurons and M2-like macrophages in jejunum of diabetic rats. The rats were randomly distributed into six groups: normoglycemic control (N), diabetic control (D) and either normoglycemic or diabetic groups treated with quercetin-loaded microcapsules at a dose of 10 mg kg-1 (NQ10 and DQ10, respectively) or 100 mg kg-1 (NQ100 and DQ100, respectively). After 60 days, the jejunum was collected. Whole mounts were immunostained for Ano1, nNOS and CD206, and oxidative stress levels and total antioxidant capacity of the jejunum were measured. Diabetes led to a loss of ICC and nitrergic neurons, but increased numbers of M2-like macrophages and elevated levels of oxidative stress were seen in diabetic animals. High-dose administration of quercetin (100 mg kg-1) further aggravated the diabetic condition (DQ100) but this treatment resulted in harmful effects on healthy rats (NQ100), pointing to a pro-oxidant activity. However, low-dose administration of quercetin (10 mg kg-1) gave rise to antioxidant and protective effects on ICC, nNOS, macrophages and oxidative/antioxidant status in DQ100, but NQ100 displayed infrequent negative outcomes in normoglycemic animals. Microencapsulation of the quercetin may become promising alternatives to reduce diabetes-induced oxidative stress but antioxidant therapies should be careful used under healthy status to avoid toxic effects.

Phenotypic changes of interstitial cells of Cajal after intestinal obstruction in rat model

The objective was to study the effect of mechanical intestinal obstruction in rats on the phenotype of interstitial cells of Cajal (ICC). Healthy Wistar rats were randomly divided into sham-operation group (C), one day obstruction group (M1), two days obstruction group (M2), and three days obstruction group (M3), with 10 rats in each group. The expression of SCF mRNA and c-Kit protein in intestinal tissue was investigated by RT-PCR and immunohistochemistry. Compared with the sham-operation group, the relative expression of SCF mRNA and the expression of c-Kit protein in intestinal tissue were significantly decreased in both obstruction groups. Levels decreased gradually with the prolongation of obstruction time, and significantly decreased on the 3rd day after obstruction (P<0.05). Immunohistochemical staining of the small intestine showed that the number of ICC in the sham-operation group was the highest, and they were gradually decreased with the extension of obstruction time in the M1 to M3 groups. There was a significant difference between groups (P<0.05). Intestinal obstruction caused a decrease in the concentrations of SCF mRNA and c-Kit protein in ICC. With the prolongation of intestinal obstruction, the number of ICCs gradually decreased.

TNF-α inhibits SCF, ghrelin, and substance P expressions through the NF-κB pathway activation in interstitial cells of Cajal

Ulcerative colitis is a chronic inflammatory disease of the colon where intestinal motility is disturbed. Interstitial cells of Cajal (ICC) are required to maintain normal intestinal motility. In the present study, we assessed the effect of tumor necrosis factor-alpha (TNF-α) on viability and apoptosis of ICC, as well as on the expression of stem cell factor (SCF), ghrelin, and substance P. ICC were derived from the small intestines of Swiss albino mice. Cell viability and apoptosis were measured using CCK-8 assay and flow cytometry, respectively. ELISA was used to measure the concentrations of IL-1β, IL-6, ghrelin, substance P, and endothelin-1. Quantitative RT-PCR was used to measure the expression of SCF. Western blotting was used to measure the expression of apoptosis-related proteins, interleukins, SCF, and NF-κB signaling pathway proteins. TNF-α induced inflammatory injury in ICC by decreasing cell viability and increasing apoptosis and levels of IL-1β and IL-6. TNF-α decreased the levels of SCF, ghrelin, and substance P, but had no effect on endothelin-1. TNF-α down-regulated expressions of SCF, ghrelin, and substance P by activating the NF-κB pathway in ICC. In conclusion, TNF-α down-regulated the expressions of SCF, ghrelin, and substance P via the activation of the NF-κB pathway in ICC.

The colonic groove of the plains viscacha (Lagostomus maximus): Histochemical evidence of an abrupt change in the glycosylation pattern of goblet cells.

The ascending colon of most rodent species shows a longitudinal colonic groove that works as a retrograde transport pathway for a mixture of bacteria and mucus toward the cecum. We describe the morphology and glycosylation pattern of the colonic groove of Lagostomus maximus to analyze the role of mucins in this anatomical feature. We also studied the distribution pattern of the interstitial cells of Cajal (ICC) to evaluate their regulatory influence on gut motility. The groove originated near the cecocolic junction and extended along the mesenteric side of the ascending colon, limited at both ends by nonpapillated ridges. These ridges divided the lumen of the ascending colon into two compartments: a narrow channel and a large channel, called the groove lumen and the main lumen, respectively. The histochemical analysis showed differences in the glycosylation pattern of the goblet cells inside and outside the groove. Unlike the mucosa lining the main lumen of the colon, the groove was rich in goblet cells that secrete sulfomucins. The PA/Bh/KOH/PAS technique evidenced an abrupt change in the histochemical profile of goblet cells, which presented a negative reaction in the groove and a strongly positive one in the rest of the colonic mucosa. The anti-c-kit immunohistochemical analysis showed different ICC subpopulations in the ascending colon of L. maximus. Of all types identified, the ICC-SM were the only cells located solely within the colonic groove.

Comparison of Cajal-like cells in pelvis and proximal ureter of kidney with and without hydronephrosis

Objectives: To evaluate effects of Cajal-like cells on human renal pelvis and proximal ureter on peristalsis. Materials and Methods: 63 patients submitted to nephrectomy due to atrophic non-functional kidney associated with hydroureteronephrosis were included as study group and 30 cases with nephrectomy due to other reasons were included as control group. Samples from renal pelvis and proximal ureters were obtained and sections of 5μ form paraffin blocks of these samples were prepared; layers of lamina propria and muscularis mucosa were examined by immune-histochemistry using CD117 in order to determine count and distribution of Cajal-like cells. Results: During immune-histochemical examinations of sections, obtained from renal pelvis and proximal ureter of hydronephrotic kidneys by CD117, Cajal-like cells number determined in lamina propria and muscularis propria was statistically significantly lower compared to control group (p<0.001). Distribution of Cajal-like cells in renal pelvis and proximal tubulus was similar under examination by light microscope, and also both groups were not different from each other regarding staining intensity of Cajal-like cells by c-kit. Conclusion: Significantly reduced number of Cajal-like cells in study group compared to control group, shows that these cells may have a key role in regulation of peristalsis at level of renal pelvis and proximal ureter in urinary system.

Tumores estromales extra-gastrointestinales (EGISTs): revisión de 4 casos con análisis inmunohistoquímico / Extra-gastrointestinal stromal tumors (EGISTs): review of four cases with immunohistochemical analysis

Los tumores del estroma gastrointestinal o GISTs son neoplasias pertenecientes a los sarcomas de partes blandas y pueden localizarse en cualquier sitio del organismo. Los más frecuentes ocurren en el tracto gastrointestinal (del esófago al recto), de hecho, más del 50 por ciento se presentan a nivel gástrico. No obstante, se han descrito en otras regiones (mesenterio y omento, retroperitoneo, vesícula biliar, pared vesical y útero). Estos últimos de localización extragastrointestinal han sido publicados con el acrónimo EGIST y representan el 5 por ciento de todos los GISTs. Se presentan 4 casos de EGIST, describimos los hallazgos anatomopatológicos, los aspectos clínicos y el perfil inmunohistoquímico. Se estudiaron 32 casos de GISTs mediante análisis prospectivo y retrospectivo, de los archivos correspondientes al Servicio y IIª Cátedra de Patología de la Facultad de Ciencias Médicas, Hospitales Córdoba y Nuestra Sra. de la Misericordia, de Córdoba (Argentina), entre los años 1987 y 2008. De dicha serie, se extrajeron y analizaron cuatro casos (2 mesentéricos y 2 retroperitoneales). Todos los casos fueron pacientes de sexo femenino entre 42 y 72 años, que consultaron por dolor abdominal, dolor pélvico y pérdida de peso. El tamaño tumoral osciló entre 4 y 28 cm. Todas las masas tumorales eran parcialmente quísticas, exhibiendo hemorragia y necrosis, en grado variable. Microscópicamente, se observaron los 2 patrones: fusocelular y epitelioide, de aspecto laxo, dispuestos en haces entrelazados con bajo índice mitótico. Panel inmunohistoquímico: Vimentina, CD34 y CD117: Positivos. Actina, cromogranina y S-100: Negativos. Ki 67: Positivo del 2 al 10 por ciento. Diagnóstico: Tumor estromal extragastrointestinal de tipo borderline. La principal peculiaridad de los casos fue su localización. Los criterios diagnósticos de certeza deben considerar, en conjunto: los aspectos clínicos, histopatológicos convencionales y el perfil inmunohistoquímico.

Gastrointestinal stromal tumors or GISTs are neoplasms belonging to the soft tissue sarcomas and can be located anywhere in the body. They most frequently occur in the gastrointestinal tract (esophagus to rectum), in fact, over 50 percent occur at the gastric level. However, they have also been described elsewhere (mesentery and omentum, retroperitoneum, gall bladder, bladder wall and uterus). Extragastrointestinal latter location have been published by the acronym EGIST (representing 5 percent of all GISTs. We report four cases of EGIST (2 of mesenteric and retroperitoneal location) describe the histopathological, clinical aspects and immunohistochemical profile. We studied 32 cases of GISTs in a prospective and retrospective analysis of files for the Service and IInd Chair of Pathology (Emergency Municipal Hospital), Hospital Córdoba, and Our Lady of Mercy, Cordoba (Argentina), between 1987 and 2008. In that series were extracted and analyzed four cases (two retroperitoneal and two mesenteric). All cases were female patients whose ages ranged from 42 to 72 years who consulted for abdominal pain, pelvic pain and weight loss. Tumor size ranged from 4 to 28 cm. All were partially cystic mass, showing hemorrhage and necrosis, in varying degrees. Microscopic patterns were observed 2 spindle and epithelioid-looking lax, arranged in bundles intertwined with low mitotic index. Immunohistochemistry panel: Vimentin, CD34 and CD117: Positive. Actin, chromogranin and S-100: Negative. Ki 67: + 2 to 10 percent. Diagnosis: extragastrointestinal stromal tumor, borderline. Complete histopathological and immunohistochemical panels should be considered for a definite diagnosis. The main peculiarity of the present cases is their location. The EGIST are rare (5 percent of GIST).

Depopulation of interstitial cells of cajal in chagasic megacolon: towards tailored surgery? / Depleção de células intersticiais de Cajal no megacólon chagásico: a caminho do tratamento cirúrgico individualizado?

BACKGROUND: The mechanism of constipation in patients with Chagasic megacolon remains partially explained. In these patients, it was recently demonstrated a reduction in the population of interstitial cells of Cajal. AIM: To evaluate density of Cajal cells in the surgically resected colon of Chagasic patients in comparison to control patients, and to verify possible association between preoperative and postoperative bowel function of Chagasic patients and colonic cell count. METHOD: Sixteen patients with Chagasic megacolon were operated on. Clinical pre- and post-operative evaluation using the Cleveland Clinic Constipation Score was undertaken. Resected colons were examined. Cajal cells were identified by immunohistochemistry using anti-CD117 antibody. The mean cell number was compared to resected colons from 16 patients with non-obstructive sigmoid cancer. Association between pre-and post-operative constipation scores and cell count for megacolon patients was evaluated using the Pearson coefficient correlation test (r). RESULTS: A reduced number of Cajal cells [cells per field: 2.84 (0-6.6) vs. 9.68 (4.3-13) - p<0.001] was observed in the resected colon of Chagasic patients when compared to colon cancer patients. No correlation between constipation score before (r=-0.205; p=0.45) or after surgery (r=0,291; p=0.28) and cell count for megacolon patients was observed. CONCLUSIONS: Patients with Chagasic megacolon display marked reduction of interstitial cells of Cajal when compared to non-Chagasic patients. An association of constipation severity and Cajal cells depopulation in Chagasic megacolon could not be demonstrated.

RACIONAL: A fisiopatologia da constipação intestinal nos pacientes portadores de megacólon chagásico permanece parcialmente esclarecida. Recentemente demontrou-se que nesses pacientes, o contingente de células intersticiais de Cajal está reduzido assim como ocorre em outros distúrbios funcionais gastrointestinais. OBJETIVO: Avaliar a densidade de células intersticiais de Cajal no intestino ressecado de pacientes submetidos a tratamento cirúrgico eletivo de megacólon chagásico em comparação com a observada no cólon de pacientes controles, e verificar possível associação entre o grau de constipação intestinal de pacientes com megacólon chagásico no pré e no pós-operatório e o grau de despopulação de células de Cajal. MÉTODO: Dezesseis pacientes com megacólon chagásico foram operados. A avaliação da função intestinal através do escore de constipação da Cleveland Clinic foi registrada antes e após a operação. O espécime cirúrgico foi examinado e as células de Cajal foram identificadas pela técnica imunoistoquímica empregando o anticorpo anti-CD117 (c-kit). O número médio de células de Cajal nos megacólons foi comparado com o de cólons ressecados de 16 pacientes [7 mulheres; média de idade de 62,8 (23-84)] operados de câncer não-obstrutivo do sigmóide. A associação entre o escore de constipação no pré e no pós-operatório e a contagem de células de Cajal foi medida através do coeficente de correlação de Pearson (r). RESULTADOS: Redução significativa no número de células de Cajal (células por campo de 400X: 2,84 (0-6,6) vs. 9,68 (4,3-13) - ] foi observada nos megacólons quando comparados ao espécime de pacientes operados por câncer. Não foi observada associação entre o escore de constipação pré (r=-0.205; p=0.45) ou pós-operatório (r=0,291; p=0.28) a a contagem de células de Cajal para os pacientes com megacólon. CONCLUSÕES: Pacientes com megacólon chagásico exibem acentuada redução no número de células intersticiais de Cajal no intestino grosso...