The effects of midazolam and ketamine on D-tubocurarine-induced twitch depression in septic rat diaphragm

The aim of this study was to elucidate the effects of midazolam and ketamine on neuromuscular blockade induced by non-depolarizing muscle relaxants (NDMRs) under the condition of sepsis induced by panperitonitis. A CLP operation (laparotomy, cecal ligation, and puncture of the cecum; septic group) or sham laparotomy (sham group) was performed on rats under O2-isoflurane anesthesia. At 18 hours after the operation, isometric twitch tensions of rat nerve-hemidiaphragm preparations elicited by indirect or direct stimulation at 0.1 Hz were measured. Midazolam enhanced the dTc (1 microM)-induced twitch depression (p < 0.05) at a high concentration (10 microM) in the septic group but not in the sham group. Ketamine enhanced the dTc (1 microM)-induced twitch depression in the sham group (p < 0.01) but not in the septic group. Midazolam and ketamine had no effect on directly elicited twitch tensions in either group. The results indicate that sepsis facilitates the midazolam-induced enhancement of the neuromuscular blocking effect of dTc but, conversely, inhibits the ketamine-induced enhancement. Sepsis elicits manifold alterations in the influence of intravenous anesthetics and sedatives on NDMR-induced neuromuscular blockade.     

Influence of the parameters of several pulse sequences on the image contrast: comparison between the two major contrast media

We compared the influences of the parameters of several pulse sequences using two major commercially available gadolinium (Gd) contrast media for MR imaging. The phantom of Gd solutions of various concentration (0.1 - 10mmol/L) was prepared, and was scanned with a 1.5T clinical MR unit, using a spin-echo T1-weighted sequence, 2DFLASH, 3DFLASH, and 3DVIBE. The signal intensity was measured and the contrast enhancement ratio (CER) was calculated and plotted as a function of Gd concentration. The results were compared between the pulse sequences, and between the contrast media as well. Both 3DFLASH and 3DVIBE showed higher CER than other two sequences, showing similar CER curve configuration. There was no significant difference both in CER value and CER curve configuration between the two contrast media for each pulse sequence.     

Lokalisierung absteigender Atmungsbahnen im Rückenmark der Katze mittels antidromer Reizung

Zusammenfassung 1. Sowohl inspiratorische als auch exspiratorische Nervenzellen innerhalb der gleichbenannten Areale der Medulla oblongata rostral und caudal des Obex senden direkte Axone herunter ins Rückenmark.2. Die Mehrzahl der absteigenden inspiratorischen Fasern kreuzt in der unteren Medulla oblongata auf die Gegenseite, der Rest verläuft ipsilateral. Die ungekreuzten inspiratorischen Fasern finden sich diffus verteilt im ventralen Teil des Seitenstranges und dorsalen Teil des Vorderseitenstranges. Die exspiratorischen Fasern kreuzen fast alle und ziehen dann im Tractus reticulospinalis des Vorderseitenstranges der Gegenseite caudalwärts. Bilaterale Axon-Kollateralen fanden sich nie.3. Die durchschnittliche Leitungsgeschwindigkeit der absteigenden respiratorischen Fasern des Rückenmarkes beträgt 40 m/sec.4. Inspiratorische R-- und R--Neurone haben direkte spinale Projektionen.5. Inspiratorische Nervenzellen, deren Axone ipsilateral descendieren, können in der Medulla oblongata unmittelbar benachbart zu inspiratorischen Nervenzellen mit kontralateral descendierenden Axonen sein.

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1. Inspiratory and expiratory nerve cells located in the corresponding zones within the medulla oblongata rostral and caudal to the obex have their axons descending directly in the spinal cord.2. The majority of the descending inspiratory fibres cross in the lower part of the medulla oblongata to the opposite side; the rest descends ipsilaterally. Of the crossed and uncrossed inspiratory fibers, most are found diffusely distributed throughout the ventral side of the lateral column and the dorsolateral part of the ventral column. Almost all expiratory fibers cross in the lower part of the medulla oblongata and then descend caudally with the tractus reticulospinalis situated in the dorsolateral part of the ventral column. We found no bilateral axon collaterals of any respiratory neurone.3. The main conduction velocity of the descending respiratory fibers of the spinal cord was 40 m/sec.4. Inspiratory R-- as well as R--neurones have spinally descending axons.5. Inspiratory nerve cells with ipsilaterally descending axons are situated in the intimate neighbourhood of those with contralaterally descending axons.

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Mit 8 Textabbildungen

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Mit Unterstützung der Deutschen Forschungsgemeinschaft und der Göttinger Akademie der Wissenschaften.     

Induction of hRAD9 is required for G2/M checkpoint signal transduction in gastric cancer cells.

DNA damage triggers the activation of checkpoints that delay cell cycle progression to allow for DNA repair. Loss of G2 checkpoints provides a growth advantage for tumor cells undergoing aberrant mitosis. However, the precise mechanisms of G2 checkpoints acting in gastric cancer are unknown. Here, we analyzed the G2 checkpoint function in two gastric cancer cells, MKN-28 cells containing a mutant p53 gene and MKN-45 cells which have wild-type p53. Two agents damaging DNA, camptothecin (CPT) or ultraviolet light (UV), were utilized to trigger a G2 phase cell cycle checkpoint response in these tumor cells. Both CPT and UV inhibited the growth of MKN-45 cells, whereas they did not affect the growth of MKN-28 cells. CPT induced cell cycle arrest at the G2/M phase and enhanced the expression of human RAD9 (hRAD9) in MKN-45 cells. In addition, hRAD9 showed perinuclear staining and similar localization with Bcl-2 in MKN-45 cells but not in MKN-28 cells after having applied CPT or UV light. These results suggest that besides p53 activity, the induction of hRAD9 is required for G2/M checkpoint signal transduction in gastric cancer cells.     

IMMUNOELECTRON MICROSCOPIC LOCALIZATION OF THYROGLOBULIN IN HUMAN FOLLICULAR ADENOMA

An electron microscopic immunohistochemical localization of thyroglobulin (TG) using PAP methods has been made in 15 cases of cold follicular adenoma. All cases of follicular adenoma showed organ specific functions such as synthesis, storage, reabsorption, and hydrolysis of thyroglobulin except for an area composed of follicular cells with trabecular arrangement. Immuno-reaction product for TG was precisely demonstrated in follicular lumina, subapical vesicles and reabsorbed colloid droplets. The reaction product observed in the follicular lumen was clearly demarcated from the cytoplasm of the follicular cells by the apical plasma membrane. The subapical vesicles ranging approximately from 50 mμ to 300 mμ in diameter were rarely observed in follicular adenoma and some of them fused with the reabsorbed colloid droplets. The reabsorbed colloid droplets usually had the intense reaction product and hydrolyzed colloid droplets had a vacuole containing floccular low electron dense materials. There is no reaction product in rough endoplasmic reticulum and Golgi complexes.     

Analysis of proteins in urinary tract stones and urine of urolithic patients

In order to investigate the mechanism of urinary tract stone formation, we analyzed protein components in urine and the stone. Urinary proteins of healthy subjects and urolithic patients as well as protein components urinary tract stone of the urolithic patients were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Electrophoretic patterns of urinary proteins of the patients differed from those of healthy subjects after separating protein patterns into those larger than 66kDa or smaller than 30kDa. Protein constituents of urinary tract stone were mainly separated into 18 bands ranging from 26.8 to 143 kDa. Major bands among these 18 bands differed among stones from different patients. On western blotting, the developed intensities of Tamm-Horsfall protein (THP) were fainter than those of healthy subjects. Whereas intensities of albumin (ALB) were stronger than those of healthy subjects. Moreover, blotting patterns of THP of the patients on non-reducing SDS-PAGE were obviously broad. Thus, we suggest that analysis of fractionated urinary proteins or protein components of urinary tract stone may provide a tool for monitoring the prognosis or relapse in the patients.     

In situ hydrogelation of photocurable gelatin and drug release.

We devised an in situ tissue-adhesive, drug-release technology based on a photoreactive gelatin, which allows in situ drug-incorporated gel formation on living tissues and sustained drug release directly on diseased tissues. Styrene-derivatized gelatins, synthesized by condensation reaction of gelatin with 4-vinylbenzoic acid, were photopolymerized in the presence of a water-soluble camphorquinone derivative as a photoinitiator upon visible-light irradiation to form swollen gels. Using albumin as a drug model, gelation characteristics and drug-release characteristics easily were manipulated by material variables, formulation variables, and operation variables. Tissue adhesivity of the gel was superior to that of fibrin glue. The biologic response, which was evaluated by intraperitoneal implantation in rats, showed that the gel was biodegraded and biosorbed, without cytotoxicity, within a few months after implantation. An in situ processable tissue-adhesive local drug release system effectively may be used to help inhibit tumor recurrence.     

Anin vitro invasion model for human renal cell carcinoma cell lines mimicking their metastatic abilities

We developed a modifiedin vitro invasion assay system using monolayers of vascular endothelial cells. A type I collagen gel was formed in plastic dishes, and overlaid with type IV collagen. Calf pulmonary arterial endothelial (CPAE) cells were seeded onto these plates, and incubated until they reached confluence. Five human renal cell carcinoma cell lines with various metastatic potentialsin vivo were then seeded on the monolayer CPAE cells, and their colony formation and invasion activities were examined for 9 days. At day 4, the highly metastatic cell lines increased the number of colony foci on monolayer CPAE cells several fold higher than their poorly metastatic counterpart. The horizontal spreading patterns were also different between poorly and highly metastatic cell lines. On day 9, the number of carcinoma foci that penetrated the monolayer of CPAE cells and type IV collagen sheets into type I collagen gels in highly metastatic cell lines greatly increased as compared with that of poorly metastatic cell lines. Ourin vitro invasion assay using monolayer CPAE cells would be useful to evaluate protease activities and colony formation during invasion.     

The number of proliferating cell nuclear antigen positive cells in endometriotic lesions differs from that in the endometrium

Immunohistochemical expression of proliferating cell nuclear antigen (PCNA) was studied in the endometrium and in endometriotic lesions during the menstrual cycle and in post-menopausal patients. During the menstrual cycle, in the basal layer of the endometrium, an increase in the number of positive indices (PI) of PCNA was observed in epithelial cells from the menstrual phase. It reached a maximum in the proliferative phase and decreased in the secretory phase. However, no change was observed in the stromal cells of the basal layer. In the functional layer of the endometrium, the PI of the epithelial cells showed a high peak in the late proliferative phase, decreased sharply in the secretory phase and remained unchanged thereafter. The PI of the stromal cells in the functional layer showed two peaks, one in the late proliferative and the other in the mid and late secretory phase. In the endometriotic lesions, except for the proliferative phase, the number of PI was significantly higher than that of the corresponding endometrium and no significant changes were observed during the menstrual cycle. In post-menopausal endometriotic lesions, the number of PI was also higher than that of the corresponding endometrium. Thus the numbers of PI differed between the endometrium and endometriotic lesions in the same patients. These results imply that the endometriotic lesions are constantly more proliferative than the endometrium irrespective of the hormonal milieu during both the menstrual cycle and in a post-menopausal environment.