Diagnostic value of dilated intercellular space for non-erosive reflux disease / 中华消化内镜杂志

Objective To explore the diagnostic value of dilated intercellular space detected by light microscope for non-erosive reflux disease (NERD) and erosive esophagitis (RE). Methods A total of 104 subjects were divided into normal control group (n = 20), NERD group (n = 30) and RE group (n = 54).Biopsies were taken at 2-3 cm above the dentate line and were examined by light microscope to calculate the intercellular space and compared between different groups. Results The mean values of intercellular space in RE ( 1.40 ±0. 17 μm) and NERD ( 1.11 ± 0. 14 μm) were significantly higher than that in control group (0.66±0. 18 μm, x2 = 154. 170, P =0.000). But no significant difference was noted between RE and NERD groups ( t = 0. 044, P = 0. 834). The cut-off value of mean intercellular space with light microscope was 0. 89 μm, with sensitivity and specificity at 95.2% and 95.0%, respectively. Conclusion Dilated intercellular space under light microscope can be a sensitive, specific and objective indicator of NERD.

Microscopic measurement of intercellular space of squamous epithelium in lower part esophagus of patients with gastro-esophageal reflux disease / 中华消化内镜杂志

Objective To evaluate the clinical significance of intercellular space diameters (ISD)of squamous epithelium by light microscopy (LM) in lower esophagus of erosive reflux esophagitis (ERD),non-erosive reflux disease ( NERD), Barrett esophagus (BE) and healthy controls. Methods A total of 21 ERD and 21 NERD patients with reflux symptoms and confirmed with 24-hour esophageal pH monitoring, 13 BE patients diagnosed by gastroscopy and biopsy, and 20 other healthy controls were enrolled in the study.Samples of ERD, NERD and control group were collected at 2 cm above dentate line, and made HE slides in the conventional way. Images for measurement of ICS were acquired with oil lens ( × 1000). ICS of squamous epithelium was quantitatively measured by computer-assisted morphometry. Ten cells were taken for each sample, 10 consecutive ISD for each cell, i.e. 100 ISD for each subject. Mean ISD was calculated.Results Mean ISDs by LM in control, BE, ERD, and NERD groups were 0. 59, 0. 99, 1.29 and 1.06 μm, respectively. The mean ISDs in BE, ERD, and NERD group were much greater than that in control (P＜0. 05). The mean, maximal and minimal ISDs of group ERD were greater than those of NERD and BE (P = 0. 000). However, the ISDs of NERD and BE are of no significant difference ( P ＞ 0. 05 ). The cut-off value of mean ISD for diagnosis of gastro-esophageal reflux disease (GERD) was 0. 85 μm. Diagnostic sensitivity and specificity for ERD, NERD and BE were 89. 1% and 100. 0%, with reference to clinical symptoms, endoscopy and ISDs above the cut-off value. Conclusion Larger ISDs in lower esophagus by using LM will be found in all subgroups of GERD, including ERD, NERD and BE. Increased ISDs may be one of the markers for diagnosis of ERD, NERD and BE.

ß (1,3) glucanaaas de los espacios intercelulars de hojas de tomate lycopersiccon esculentum ceraciforme después de infección con phytophthora infestans / ß (1,3)glucanasee from intercelluar space of tomato leaves (lycopersicon esculentum ceraciforme) after infección with phytophthora infestans / ß (1,3) gluanasaas dos espacios intercelulaes de folhas de tomate lycoperssicon esculentum ceraciforme depois de infeccao com phytophthora infestans

En este trabajo se analizó el líquido de lavado intercelular de Lycopersicon escu-lentum ceraciforme L1568 perteneciente al germoplasma nacional con resistencia en campo a P. infestans el cual mostró la presencia de cuatro ß (1,3) glucanasas después de infección con el patógeno. Para tal efecto las plantas se inocularon con el patógeno con el fin de inducir aumento en la concentración de las proteínas PR y otras sustancias relacionadas con defensa; 15 días después de la inoculación con el parásito, se extrajo el líquido de lavado intercelular de las hojas de tomate utilizando agua. Las proteínas con actividad de ß (1,3) glucanasa extraídas se liofilizaron, se resuspendieron en búfer, se purificaron utilizando cromatografía de exclusión, intercambio iónico y electroforesis preparativa. Los resultados de caracterización parcial, mostraronlaexistenciadeuna ß (1,3) glucanasa básica con peso molecular de 36,8 kDa y pI 9,2 y tres ß (1,3) glucanasas ácidas de 35,4; 30,1 y 7,2 kDa con puntos isoeléctricos de 3,8; 3,6 y 5,0 respectivamente. Adicionalmente se determinaron las propiedades cinéticas de cada enzima, KM, Vmáx,Eo yK3 encontrando, de acuerdo con los resultados obtenidos, que la proteína básica no ha sido reportada en la literatura relacionada con tomate.

In this paper was analysed the intercellular washing fluid of Lycopersicon esculentum ceraciforme L1568 belonging to the national germplasm with resistance to P. infestans in field which showed the presence of four ß (1,3) glucanases after infection with the pathogen. For this purpose the plants were inoculated with the pathogen to induce a concentration increase of PR proteins and other defence-related substances, 15 days after inoculation with the parasite was extracted the intercellular washing fluid of tomato leaves using water. The proteins with ß(1,3) glucanase activity were extracted, freeze-dried and re-suspended in buffer solution and were purified using exclusion chromatography, ion exchange chromatography and preparative electrophoresis. The results of partial characterization showed the presence of a basic ß (1,3) glucanase with a molecular weight of 36.8 kDa and pi 9.2 and three ß (1,3) glu-canase acid 35.4, 30.1 and 7.2 kDa with isoelectric points of 3.8, 3.6 and 5.0 respectively. Additionally it was determined the kinetic properties of each enzyme, KM,Vmax, and Eo,K3, according to the results, the basic protein has not been reported in the literature related to tomato.

Este trabalho analisou o líquido de lavado intercelular de Lycopersicon esculentum ceraciforme L1568 pertencente ao ger-moplasma nacional com resistência em campo a P. infestans, mostrau a presença de quatro ß (1,3) glucanase despois a infecção com o agente patogênico. Para este efeito as plantas foram inoculadas com o agente patogénico para induzir um aumento na concentração das proteínas PR y outras substâncias relacionadas com defesa, e 15 dias mais tarde se extraiu o líquido de lavado intercelular de folhas de tomate utilizando água. As proteínas extraídas foram liofilizadas, resuspendidas em solução tampão, purificadas utilizando cromatografia de exclusão, de intercambio iónico e electroforesepreparativa,. Os resultados da caracterização paracial revelou a existência de uma ß(1,3) glucanasa basica com peso molecular de 36,8 kDa e PI 9,2, e três ß(1,3) glucanasas ácidas de 35.4, 30.1 e 7.2 kDa com pontos isoeléctricos de 3.8, 3.6y 5.0, respectivamente. Adicionalmente, foi possível determinar as propriedades cinéticas KM,Vmax,Eo yK3 de cada enzima, encontrando de acordo com os resultados que a proteína básica não havia sido reportada na literatura.relacionada com tomate.

A study on the light microscopic measurement of intercellular space of squamous epithelium in lower-esophagus to diagnose gastroesophageal reflux disease / 中华内科杂志

Objective To evaluate the significance of dilated intercellular space (DIS) of squamous epithelium in lower-esophagus which was measured at light microscopy (LM) in the diagnosis of gastroesophageal reflux disease (GERD). Methods This study was divided into two parts. The first part consists of 133 GERD patients with typical symptoms, including 75 erosive esophagitis (EE) and 58 non-erosive reflux disease (NERD);the second part consists of 25 healthy volunteers as control; ambulatory 24-hr esophageal pH monitoring, endoscopy and biopsies which were taken in lower esophagus were performed in all individuals. NERD patients with negative ambulatory 24-hr esophageal pH monitoring were undertaken PPI test. We selected 43 individuals randomly (4 control, 11 NERD and 28 EE) to measure the intercellular space diameter with LM and transmission electronic microscopy (TEM) simultaneously. All the samples were observed at immersion objective, taken pictures and make the scale at the same time; all the pictures were measured by the computer-assisted morphometry and in the samples for one ease should be measured 100 points and then calculate the mean intercellular space diameter, while the TEM has the same procedures. Results A total of 158 individuals (90 male, 68 female) were enrolled in this study. The mean intercellular space diameter with LM of the controls, the NERD and the EE patients were (0.61±0.10)μm, (1.12±0.61)μm and (1.30±0.19) μm, respectively, with significant differences between the control and NERD group, the control and EE group, the EE and NERD group (P <0.05); the mean intercellular space diameter with LM of EE subgroup of LA-A, LA-B, LA-C, LA-D were (1.31±0. 23)μm, (1.27±0.17)μm, (1.31±0.14)μm, (1.33±0.11)μm, respectively, without significant differences among every two subgroups (P > 0.05);NERD patients with positive and negative 24-hr ambulatory pH monitoring were 40 and 17 cases, and the mean intercellular space diameter with LM were (1.12±0.16) μm, (1.11±0.31) μm, respectively, without significant differences between them (P >0.05) ;the cut-off value of mean intercellular space diameter with LM was 0. 85 μm, if combination the clinical GERD symptom, endoscopy and 24-hr ambulatory pH monitoring as gold standard, its sensitivity was 95.5% and specificity was 100%;the intercellular space diameter of the controls, the NERD patients and the EE patients with TEM were (0. 30±0.14)μm, (1.33±0.28)μm, (1.40±0. 22)μm, respectively,with significant differences between the control and NERD group, the control and EE group (P < 0.05). It was significant correlation in intercellular space diameter between LM and TEM (r=0.737, P=0.000).Conclusions The intercellular space diameter of squamous epithelium in lower esophagus of GERD patients can make quantitative study with LM. It is significant correlation with TEM. DIS with LM is one of sensitive, specific and objective marker of GERD.

Variation of the intercellular space in the esophageal epithelium in response to hydrochloridric acid infusion in patients with erosive esophagitis

The purpose of this study was to compare esophageal infusion with 0.1 N hydrochloridric acid (HCl) to esophageal infusion with saline in patients presenting with typical gastroesophageal reflux symptoms and erosive esophagitis. METHODS: Upper gastrointestinal endoscopy was performed on 44 prospective subjects, 29 of whom were included in the study. Eighteen patients presented with normal esophagi (Control Group "C"), nine of whom were infused with HCl and nine with saline. Eleven patients presented with erosive esophagitis (Lesion Group "L"), five of whom were infused with HCl and six with saline. Biopsies of the esophageal mucosa were collected before and after infusions. RESULTS: No statistically significant difference was found between the two types of infusions in terms of the dilation of the intercellular space of the esophageal epithelium, regardless of the status of the patient. CONCLUSIONS: Response to HCl infusion cannot be used as a marker for gastroesophageal reflux disease.

PROTEÍNAS ANTIFÚNGICAS CONTRA P. INFESTANS EN LOS ESPACIOS INTERCELULARES DE HOJAS DE TOMATE (Lycopersicon esculentum cerasiforme) POSIBLES DEFENSINAS DE PLANTAS / ANTIFUNGAL PROTEINS AGAINST P. INFESTANS FROM INTRACELLULAR SPACE OF TOMATO LEAVES (Lycopersicon esculentum cerasiforme) POSSIBLE DEFENSINS OF PLANTS

Se aislaron y caracterizaron parcialmente proteínas antifúngicas de los espacios intercelulares de hojas de tomate Lycopersicon esculentum cerasiforme, variedad que ha mostrado resistencia en campo a Phytophthora Infestans; se observó que después de inoculación con el patógeno dichas proteínas se acumularon sistémicamente en la planta. Las proteínas identificadas mostraron características de Defensinas de plantas, una nueva familia de proteínas con bajo peso molecular, carga positiva a pH fisiológico y actividad antifúngica evaluada in vitro contra P. infestans. Los análisis electroforéticos en geles de poliacrilamida con SDS-Tricina en condiciones reductoras y no reductoras, sugirieron que están asociadas en trímeros y tetrámeros y poseen pesos moleculares de 5,2 kDa.

Antifungal Proteins from intercellular space of Tomato leaves Lycopersicon esculentum cerasiforme were isolated and partially characterized; this variety had shown resistance against Phytophthora infestans, after inoculating plants with the pathogen it was observed that these proteins were accumulated systemically. The isolated proteins be haved as plant defensins, a novel protein family, with low molecular weight, cationics at physiological pH and Antifungal Activity in vitro evaluated against P. infestans. Electrophoretic analyses in SDS Tricine-PAGE, under reducing and non-reducing conditions suggested that, they were associated as trimers and tetramers; and have molecular weights of 5,2 kDa.

Morphological Characteristics and Intercellular Connections of Corneal Keratocytes

PURPOSE: To investigate the morphological characteristics of keratocytes and the interconnection of keratocytes with adjacent keratocytes using the flat preparation method and scanning electron microscopy with a frontal section of the human corneal stroma. METHODS: The thin, corneal collagen lamellae were carefully dissected from the cornea (n=7), which had been stained by the flat preparation method. The remaining tissue was fixed in 3% glutaraldehyde and observed by transmission electron microscopy following the frontal section. RESULTS: The flat preparation revealed the corneal fibroblasts between the lamellae of the collagen fibers and showed that the ramifying cellular processes of the keratocytes were in contact with the cytoplasmic processes or cell bodies of neighboring fibroblasts. Two types of discrete subpopulations of keratocytes were identified: a smaller, cellular type of keratocyte with spindle-shaped nucleus with heterochromatin, and a larger, cellular type with a large indented nucleus with relatively scanty cytoplasm. Collagen fibers ran parallel to each other toward the fenestration of the cytoplasmic wall of the keratocyte. CONCLUSIONS: These flat preparation method results showed that the keratocytes within the corneal stroma are interconnected with the adjacent keratocytes, which indicates the presence of a functional communicating network through the keratocyte circuits within the stroma. A smaller, cellular type of keratocyte with spindle-shaped nucleus was morphologically differentiated from a larger, cellular type with a large, indented nucleus by flat preparation and transmission electron microscopy.