Isolation, growth, and characteristics of human ovarian surface epithelium

To isolate human ovarian surface epithelium (OSE) and studied its characteristicsin vitro, we used the scraping method with an enzymatic procedure. After incubation of the ovary in collagenase type 1 solution, the surface cells were removed by gentle scraping with surgical blade. Cells obtained as a cluster after unit gravity sedimentation with 5% bovine serum albumin in medium 199 were cultured in medium 199 containing 15% fetal bovine serum. The viable cell number in a single ovary was 0.1 to 2.7×10⁶, and the cell population doubling time was between 7 to 10 days. Confluent monolayers were formed after 13–20 days and subcultured one to three times. The monolayers mostly had a cobblestone appearance, and fusiform or polygonal cells were also observed. By cytochemistry, immunocytochemistry and scanning and transmission electron microscopy, the cells in short-term culture were shown to have characteristics of mesothelial OSE cells.     

The characterization and regional distribution of neuromedin N-like immunoreactivity in rat brain using a highly sensitive and specific radioimmunoassay. Comparison with the distribution of neurotensin

Neuromedin N is a hexapeptide that shares a 4 amino acid homology with the C-terminus of neurotensin and exhibits neurotensin-like effects in the central nervous system. Both peptides were recently shown to be encoded in the same precursor molecule. In this study, a radioimmunoassay for neuromedin N was developed using monoiodo [¹²⁵I-Tyr⁴]neuromedin N as the tracer and a rabbit antiserum raised against synthetic [Cys⁶]neuromedin N coupled to ovalbumin through its Cys residue. The antiserum showed strong structural requirement for the N-terminal sequence of neuromedin N and did not cross-react with neurotensin and other related peptides. The limit of detection of the radioimmunoassay was 0.5 fmol/tube and the IC₅₀ was 5 fmol/tube. Neuromedin N-like immunoreactivity was present in 0.1 N HCl extracts of rat brain at a concentration of 9.3 ± 1.3pmol/g of tissue and behaved like synthetic neuromedin N on HPLC. Its concentration was significantly lower than that of neurotensin assayed in the same extracts (15.1 ± 1.4pmol/g, and this was not the consequence of lower extraction yield or lower post-mortem stability of neuromedin N as compared to neurotensin. The regional rat brain distribution of neuromedin N-like immunoreactivity paralleled that of neurotensin-like immunoreactivity, being highest in the hypothalamus and lowest in the cerebellum. These data support the proposal of a neuromodulator role for neuromedin N. The highly specific and sensitive radioimmunoassay described here will make it possible to investigate in more detail the regional brain distribution of neuromedin N and to study its release from brain tissues.     

Characterization of calcium fenoprofen 1. Powder dissolution rate and degree of crystallinity

Samples of calcium fenoprofen crystals have been prepared on laboratory, pilot and production scales, some by conventional aqueous precipitation, others under conditions designed to increase or decrease the degree of crystallinity. They were characterized by X-ray powder diffraction, scanning electron microscopy, Fourier transform infrared spectroscopy, surface area by nitrogen sorption, agglomerate size by Coulter counter, true density, sodium content, powder dissolution rates and heats of solution. No evidence of polymorphic variation was found. Most precipitation conditions gave partially fused agglomerates of primary crystals. Relative degrees of crystallinity were assessed from heats of solution. The more perfectly crystalline samples gave relatively high endothermic heats of solution coupled with low powder dissolution rates. Lattices with high levels of disruption, or low crystallinity, gave lower heats of solution coupled with enhanced powder dissolution rates. Heats of solution make a significant contribution to the overall characterization and to understanding batch-to-batch variation, and they relate well to the observed powder dissolution rates.     

Preparation and in vitro/in vivo evaluation of metformin hydrochloride rectal dosage forms for treatment of patients with type II diabetes

Background: Metformin hydrochloride (MtHCL) is an oral antidiabetic drug and has many other therapeutic benefits. It has poor bioavailability, narrow absorption window and extensive liver metabolism. Moreover, children and elders face difficulty to swallow the commercial oral tablets.

Objectives: Preparation, in vitro/in vivo evaluation of MtHCL suppositories for rectal administration to solve some of these problems.

Methods: Suppository fatty bases (Witepsol^®, Suppocire^® and Massa^®; different grades) and PEG bases 1000, 4000 and 6000 (different ratios), were used to prepare rectal suppository formulations each containing 500?mg drug. These were characterized for manufacturing defects, and pharmacotechnical performance and formulations showing superior results were subjected to bioavailability testing in human volunteers compared with the commercial oral tablet (Ref) applying LC–MS/MS developed analytical technique.

Results: The preparation method produced suppositories with satisfactory characteristics and free of manufacturing defects. The fatty bases were superior compared with PEG bases regarding the physical characteristics. Three formulations were chosen for bioavailability testing and the results showed comparable bioavailability compared to the Ref.

Conclusions: The fatty bases showed superior characteristics compared with the PEG bases. MtHCL formulated in selected fatty bases could be a potential alternative to the commercial oral tablets particularly for pediatric and geriatric patients.     

吲哚美辛肠溶滴丸分散状态的表征

摘 要 目的：考察吲哚美辛（IDM）肠溶滴丸中IDM存在的状态及与基质聚乙二醇6000（PEG 6000）的相互作用。方法: 分别采用差示扫描量热法（DSC）、红外光谱法（IR）、X-射线衍射法及扫描电镜（SEM）观察对IDM、PEG 6000、两者的物理混合物及IDM肠溶滴丸进行研究。结果: DSC图谱中,IDM肠溶滴丸在164℃处的吸热峰消失,而57℃处的PEG 6000吸热峰发生了前移;IR图谱中,IDM肠溶滴丸中IDM的羰基伸缩振动峰消失,而在1 680 cm^-1处出现了新峰;X-射线衍射图谱中,IDM肠溶滴丸无衍射峰存在;SEM下IDM以更细小的状态分布于基质中。     

A continuous cell line (KK-2) from a supratentorial primitive neuroectodermal tumor

Summary Tumor tissue located in the occipital lobe with hemorrhage was obtained from a 19-year-old patient. Histological examination indicated it to consist of undifferentiated small, round cells without neuronal or glial differentiation, and possibly to be a type of primitive neuroectodermal tumor. The tumor cells were cultured for 3 years and a continuous cell line (KK-2) was established. KK-2 was transplantable to nude mice. With immunocytochemistry, neuron-specific enolase, protein gene product 9.5, vimentin, TUJ1 (a monoclonal antibody specific for neuron-associated class III -tubulin isotype) and 6H7 (a monoclonal antibody to NCAM produced by us) were detected. None of the following could be found: glial fibrillary acidic protein, S-100 protein, neurofilament and synaptophysin, calcitonin gene-related peptide, gastrin releasing peptide corticotropin-releasing factor, substance P, somatostatin, chromogranin, aromatic l-amino acid decarboxylase and tyrosine hydroxylase. The original tumor and KK-2 cells obtained after 3 years of culture and transplants in nude mice displayed essentially the same ultrastructural and immunohistochemical characteristics. KK-2 cells showed no differentiation to mature neuronal, glial or ependymal cells. This cell line may possibly serve as a useful model for studying cellular differentiation of human neuroectodermal tumors and normal neuronal development.Supported in part by a Grant-in-Aid for Cancer Research from the Ministry of Health and Welfare (1–5), Japan     

Endoscopic ultrasound and intraductal ultrasound in the diagnosis of small pancreatic tumors

Background: The purpose of this study was to assess the diagnostic value of endoscopic ultrasound (EUS) and intraductal ultrasound (IDUS) in the detection of small pancreatic tumors. Methods: EUS was performed in 166 patients with verified pancreatic disease. IDUS was performed in 46 patients. A microprobe was introduced into the main pancreatic duct through the papilla of Vater using the duodenoscope. Results: EUS was valuable in the detection of small pancreatic tumors. Ductal adenocarcinomas smaller than 1 cm were demonstrated as a hypoechoic mass with a central irregular hyperechoic area. EUS and IDUS were useful in the characterization of intraductal paillary tumors (ductectatic mucinous tumors). EUS demonstrated nodular excrescences, and IDUS depicted papillary proliferation of the duct epithelium, which are characteristic of carcinomas and adenomas but not of hyperplasia. Internal architecture of cystic neoplasms was clearly depicted by EUS, and differentiation of serous and mucinous tumors was readily achieved. A tumor as small as a 5-mm islet cell was demonstrated on EUS because islet cell tumors are very hypoechoic. Conclusion: EUS and IDUS are relatively noninvasive procedures and are useful in the detection of small tumors and differentiation of pancreatic diseases. Received: 0/0/0/Accepted: 0/0/0     

Endoscopic ultrasound elastography: current status and future perspectives

Elastography is a new ultrasound modality that provides images and measurements related to tissue stiffness. Endoscopic ultrasound(EUS) has played an important role in the diagnosis and management of numerous abdominal and mediastinal diseases. Elastography by means of EUS examination can assess the elasticity of tumors in the proximity of the digestive tract that are hard to reach with conventional transcutaneous ultrasound probes, such as pancreatic masses and mediastinal or abdominal lymph nodes, thus improving the diagnostic yield of the procedure. Results from previous studies have promised benefits for EUS elastography in the differential diagnosis of lymph nodes, as well as for assessing masses with pancreatic or gastrointestinal(GI) tract locations. It is important to mention that EUS elastography is not considered a modality that can replace biopsy. However, it may be a useful adjunct, improving the accuracy of EUSfine needle aspiration biopsy(EUS-FNAB) by selecting the most suspicious area to be targeted. Even more, it may be useful for guiding further clinical management when EUS-FNAB is negative or inconclusive. In the present paper we will discuss the current knowledge of EUS elastography, including the technical aspects, along with its applications in the differential diagnosis between benign and malignant solid pancreatic masses and lymph nodes, as well as its aid in the differentiation between normal pancreatic tissues and chronic pancreatitis. Moreover, the emergent indication and future perspectives are summarized, such as the benefit of EUS elastography in EUS-guided fine needle aspiration biopsy, and its uses for characterization of lesions in liver, biliary tract, adrenal glands and GI tract.     

Potential of electrospun chitosan fibers as a surface layer in functionally graded GTR membrane for periodontal regeneration

Objective

The regeneration of periodontal tissues lost as a consequence of destructive periodontal disease remains a challenge for clinicians. Guided tissue regeneration (GTR) has emerged as the most widely practiced regenerative procedure. Aim of this study was to electrospin chitosan (CH) membranes with a low or high degree of fiber orientation and examines their suitability for use as a surface layer in GTR membranes, which can ease integration with the periodontal tissue by controlling the direction of cell growth.