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Biomedical subjects

J Gu

Publications and source records attributed to J Gu.

359 records · Page 20Linked to original sources

Vipergic nerves in the penis.

High concentrations of vasoactive intestinal polypeptide (VIP) were detected by immunocytochemistry and radioimmunoassay in thirty surgical specimens of male external genitalia. VIP was found exclusively in fine autonomic nerves. VIPergic nerves were most densely concentrated in the penis around the pudendal arteries and in the erectile tissue of the corpus cavernosum. Considerable numbers of VIP nerve fibres were also seen in the vas deferens and epididymis. VIP is known to exert regulatory actions on blood-flow, secretion, and muscle tone. Its presence in considerable amounts in the male genital tract suggests that this newly discovered peptide neurotransmitter may be important in the nervous control of male external genitalia.

Adult↗

Sequential use of the PAP and immunogold staining method for the light microscopical double staining of tissue antigens.

Double immunoperoxidase staining using different couplers can give various combinations of colours on a single tissue section to achieve a comparable picture of different antigens. However, the colour combinations achieved to date are not entirely satisfactory. A double immunostaining procedure is introduced here, combining the peroxidase anti-peroxidase (PAP) and immunogold staining (IGS) methods. The IGS method is a new, simple, sensitive and reliable approach to immunostaining at the light microscopic level. It was carried out in three ways. Firstly, a two-step method was used in which the second layer was goat anti-rabbit IgG absorbed onto gold particles (GAR/Au20). Secondly, a three-step method was employed where the second layer was unlabelled goat anti-rabbit IgG and the third layer was a rabbit antibody to peroxidase absorbed onto the gold particles (RAP/Au20) and acting as a gold-labelled IgG antigen. The third method combined the first two methods using GAR/Au20 as th second layer and RAP/Au20 as the third layer which increased the amount of bound gold and enhanced the red colour, providing a better picture. The use of gold-labelled antibodies in double immunostaining has great potential value for many studies including that of the diffuse neuroendocrine system of the gut.

Animals↗

Neuron-specific enolase in the Merkel cells of mammalian skin. The use of specific antibody as a simple and reliable histologic marker.

Merkel cells are specialized skin receptor cells, characterized by their particular location in the epidermis and close association with nerve terminals. Although they can be distinguished ultrastructurally by their small, electron-dense secretory granules, there is no specific and reliable method for identifying them by light microscopy. Using antibodies to neuron-specific enolase (NSE), the authors have shown sparsely distributed groups of specifically immunostained cells and associated nerve terminals in the nose skin of cats and rats. These cells were easily distinguished from other epithelial cell types, including melanocytes and Langerhans cells and had all the morphologic features of Merkel cells and their so-called neurite complexes, including the characteristic cytoplasmic secretory granules (60 nm in diameter). NSE immunostaining is a simple and reliable method for the specific light-microscopic staining of Merkel cells and provides further evidence for NSE as a marker for the diffuse neuroendocrine system.

Animals↗

Comparison of detecting sensitivities of different sizes of gold particles with electron-microscopic immunogold staining using atrial natriuretic peptide in rat atria as a model.

The detecting sensitivities of different-sized gold particles were compared in the localization of atrial natriuretic peptide (ANP) in rat atria. The secondary antibodies were goat antirabbit labeled with 5, 15, 30, or 40 nm colloidal gold diluted 1:2 to 1:100 in Tris buffer. The relative quantity of alpha-ANP immunoreactivity in specific granules was determined by subtracting the number of gold particles in 1 micron 2 nongranule area from that in 1 micron 2 granule area measured with a computerized image analyzer. The optimal dilution that achieved the maximal contrast between specific and background label was influenced by the particle size. Optimal dilutions were 1:80, 1:30, 1:20, and 1:5 for 5, 15, 30, and 40 nm gold, respectively. At optimal dilutions, the maximal detecting sensitivity (MDS) was in inverse proportion to the gold particle size; however, this relationship is not entirely linear. The ratio among the MDSs of 5, 15, 30, and 40 nm gold particles was approximately 34:9:3:2. A double immunogold staining was performed to localize alpha- and beta-ANPs with 15 and 5 nm gold, respectively. Both antigens were detected in the same granules. If the ratios established from the single staining data were used, the ratio between the alpha- and the beta-ANP antigens in the same granules was approximately 2.8:1. The data obtained in this study provide a useful reference for applications of immunogold electron microscopy in a quantitative manner, particularly for double immunogold labeling.

Animals↗

Immunogold staining procedure for the localisation of regulatory peptides.

The use of protein A- and IgG-conjugated colloidal gold staining methods for the immuno-localisation of peptide hormones and neurotransmitters at light- and electron microscope level are described and discussed. Bright-field and dark-ground illumination modes have been used to visualise the gold-labelled antigenic sites at the light microscope level. Immunogold staining procedures at the ultrastructural level using region-specific antisera have been adopted to localise specific molecular forms of peptides including gastrin (G17 and G34), glucagon and pro-glucagon, insulin and pro-insulin, in normal tissue and in tumours of the gastroenteropancreatic system. Similar methods have been used to demonstrate the heterogeneity of p-type nerves in the enteric nervous system. Vasoactive intestinal polypeptide (VIP) has been localised to granular sites (mean +/- S.D. granule diameter = 98 +/- 19 nm) in nerve terminals of the enteric plexuses and in tumour cells of diarrhoeogenic VIP-producing neoplasias (mean +/- S.D. granule diameter = 126 +/- 37 nm) using immunogold procedures applied to ultraviolet-cured ultrathin sections. Co-localisation of amines and peptides in carotid body type I cells and in chromaffin cells of normal adrenal medulla and phaeochromocytomas has also been demonstrated. Advantages of the immunogold procedures over alternative immunocytochemical techniques are discussed.

Animals↗

The origin of VIP-containing nerves in the urinary bladder of rat.

The innervation of the urinary bladder is known to include a considerable number of nerves containing vasoactive intestinal polypeptide (VIP). The origin of such nerves in the bladder of rat was investigated in this study using the methods of immunocytochemistry and radioimmunoassay combined with surgical sectioning of the hypogastric and/or pelvic nerves to the bladder. Eight days after pelvic nerve sectioning proximal to the main pelvic ganglion, VIP-immunoreactive nerves and VIP content were markedly increased from the level in the sham-operated rat bladder. Sectioning of hypogastric or both nerve pathways led to a less significant increase. It was therefore postulated that the majority of VIP-immunoreactive nerves originate from ganglia located either close to the bladder or within the bladder wall. It is interesting that in these experiments the VIP content of the bladder nerves is inversely related to the changes in motility that would be expected to result from the nerve sections.

Animals↗

Calcitonin gene-related peptide (CGRP) in the female rat urogenital tract.

CGRP-immunoreactivity was found throughout the female rat urogenital tract by specific radioimmunoassay, and shown to be present in nerve fibres by immunocytochemistry. The highest concentrations of CGRP-like immunoreactivity were found in the urinary tract, with lower levels in regions of the genitalia. Chromatographic analysis of bladder and vaginal extracts on Sephadex G-50 columns and HPLC revealed at least three CGRP-immunoreactive peaks. The major peak emerged in the same position as synthetic rat CGRP. CGRP nerve fibres were associated mainly with blood vessels, non-vascular smooth muscle, squamous epithelium and uterine and cervical glands, and were particularly abundant in the ureter and bladder. CGRP-immunoreactivity was depleted by neonatal treatment with capsaicin and after surgical section of pelvic and/or hypogastric nerves. Immunocytochemistry demonstrated that depletion occurred predominantly in the mucosal layer of the urogenital tract. These findings indicate a sensory function for most of the CGRP-immunoreactive nerves in the rat urogenital tract.

Animals↗

Effect of 6-hydroxydopamine on neuropeptides in the rat female genitourinary tract.

The occurrence and distribution of neuropeptide Y has been determined in the rat female genitourinary tract by radioimmunoassay and chromatographic analysis. Within the bladder, higher concentrations of neuropeptide Y were found in the trigone (48.8 +/- 5.2 pmol/g) than in the dome (36.0 +/- 2.1 pmol/g). In the genital tract, highest concentrations were identified in the vagina (41.4 +/- 2.1 pmol/g). Treatment of rats with 6-hydroxydopamine resulted in significant depletion of neuropeptide Y concentrations in both parts of the bladder, together with vagina, uterine horn and fallopian tube. No change was observed in the cervix, uterine body and ovary. Concentrations of vasoactive intestinal polypeptide were unaffected by treatment with 6-hydroxydopamine except in the area of the cervix where concentrations rose from 64.1 +/- 5.7 pmol/g to 133.6 +/- 15.1 pmol/g (p less than 0.05). There was a generalised, but statistically insignificant rise in substance P concentrations.

Animals↗

Purification of transthyretin by high performance affinity chromatography from human plasma.

The partial purification of human transthyretin (TTR) by high performance affinity chromatography with the help of other separation techniques is described in the present paper. A new affinity medium was prepared with a monosized (ca. 10 microns particle size) macroporous resin as the support and thyroxine (T4) as the ligand. The purification of TTR was carried out in a few simple steps involving serum precipitation, anion exchange, Thyroxine affinity chromatography and gel filtration. The overall yield was 29% and the refined TTR contained less than 2% impurities as analyzed by RP-HPLC. When TTR was administrated to the culture medium DMEM of liver tumor strain SMMC-7721, a true inhibition of cell growth (ca. 50%) was observed as an actual decrease in cell number over time.

Chromatography, Affinity↗

Preservation of the latissimus dorsi muscle during cardiomyoplasty surgery.

BACKGROUND: Cardiomyoplasty surgery has been shown to be associated with damage and degeneration of the assisting skeletal muscle. The purpose of this study was to use ischemic (short-term) and thermal (long-term) preconditioning to protect the muscle during surgery and the subsequent ischemia. METHODS: Three 10-minute cycles of ischemia-reperfusion were accomplished noninvasively on goat latissimus dorsi muscle (LDM) immediately prior to surgery. In another experiment, LDM was noninvasively heat shocked for 20 minutes at 42 degrees C 24 hours prior to surgery. LDM damage was evaluated 5 days postsurgery using enzyme activities (beta-glucuronidase, beta-GLN; citrate synthase), hydroxyproline, morphology, and blood flow. RESULTS: The lysosomal enzyme, beta-GLN, was significantly increased (43%, p < 0.05) by surgical dissection and remained high in the ischemic preconditioned LDM (58%, p < 0.05) and in the heat shocked LDM (57%, p < 0.05). CONCLUSION: These findings show that these two protective protocols do not reduce the muscle damage that occurs during surgical preparation of the LDM for cardiomyoplasty.

Animals↗

A 63-kDa protein with androgen-binding activity is not from the androgen receptor.

We have found a new protein in the heart of rat and mice that can be selectively and covalently labelled with the synthetic androgen analog mibolerone. Binding is specific as it can be displaced by excess radioinert ligand. The protein is prominently expressed in liver, kidney, and heart, but not in skeletal muscle. It is water soluble and found in the cytosol. Under denaturing conditions it has a molecular weight of 63,000 and appears on two-dimensional gels with an isoelectric point of 6.3. The protein's affinity for androgen is lower than that of the androgen receptor and it is about 100-fold more abundant than the receptor in the heart. Expression of the protein is not induced by androgen. The presence of this protein in testicular feminization (tfm) mice with a genetical defect of the androgen receptor rules out that it is the androgen receptor or a portion thereof. The biological role of this protein is not yet known.

Animals↗

Effectiveness of fibrin glue in the reduction of postoperative intrapericardial adhesions.

The hemostatic properties of fibrin sealant have been well described. Previously published reports have attempted to clarify the possible role of fibrin glue in the inhibition of the formation of intrapericardial adhesions following cardiac surgery. Earlier work hypothesized that fibrin glue may reduce the severity of postoperative adhesions and that the use of autologous fibrin glue may have similar effects, without the risks that accompany homologous blood products. Six juvenile farm pigs were utilized to test this hypothesis. Conventional fibrin glue and single-donor fibrin glue were tested in open-heart surgery. This experimental model was also reexamined and found to be of significant utility in simulating adult reoperative cardiac surgery. The fibrin glue subjects were universally easier to reoperate due to fewer adhesions, as demonstrated grossly and histologically. The single-donor fibrin glue had no significant advantage on adhesion formation, when compared to the conventional fibrin glue group, but the ramifications of formulating fibrin glue in this fashion offer a significant benefit toward the complete use of autologous blood products in open-heart surgery.

Animals↗

Evaluation of the European Spondyloarthropathy Study Group (ESSG) classification criteria in a Chinese population.

OBJECTIVE: To evaluate for thefirst time in a Chinese population, the usefulness of the European Spondyloarthropathy Study Group (ESSG) criteria. METHODS: A total of 193 clinically diagnosed SpA patients were compared to 166 patients with other types of arthritis, using the parameters listed in the ESSG criteria. RESULTS: The sensitivity and specificity of the ESSG criteria in this Chinese population were high at 85.4% and 96.4% respectively. CONCLUSION: The use of ESSG criteria for classifying SpA in Chinese will not lead to significant omission of SpA patients or inclusion of patients who do not have SpA.

China↗

The regional distribution of NPY-, PHM-, and VIP-containing nerves in the human female genital tract.

The regional distributions of neuropeptide Y (NPY), peptide histidine-methionine (PHM), and vasoactive intestinal polypeptide (VIP) immunoreactivities in the human female genital tract have been estimated by specific radioimmunoassays, and their molecular forms determined by chromatography. The localisation and distribution of these three peptides was carried out by immunocytochemistry. The vagina and cervix contain high concentrations of NPY- and VIP-immunoreactive nerves, mainly localised around the vascular and nonvascular smooth muscle. VIP-containing nerves were, in addition, seen beneath the cervical and, in particular, the vaginal epithelium. A comparatively high level of immunoreactive NPY is found in the fallopian tube, mainly around the circular muscle coat. There is evidence that VIP is a neurotransmitter in the female genital tract, and these results suggest a similar role for NPY and PHM.

Adult↗

IL-2 gene therapy of advanced lung cancer patients.

We report here Phase I clinical-trial studies of retroviral-mediated interleukin-2 (IL-2) gene transfer to tumor-infiltrating lymphocytes that are re-infused to advanced lung cancer patients with pleural effusions. Ten lung cancer patients with malignant pleural effusions for whom all conventional therapy had failed were included in this Phase I protocol. Tumor infiltrating lymphocytes (TIL) from the patients were exposed to the retroviral plasmid pL(IL-2)SN containing the human IL-2 gene. Approximately 1-6 x 10(10) TIL cells transfected with IL-2 were re-infused into the chest cavity of each patient. The toxicity of this treatment with TIL/IL-2 gene therapy in these patients was minimal with transient slight fever of approximately 37.5. Pleural effusions did not re-accumulate for at least 4 weeks in six of ten patients. One patient was observed to have not only the resolution of the pleural effusions, but in addition the size of the original tumor decreased as seen by CT. The clinical results indicate this method of cancer gene therapy is safe and possibly efficacious against pleural effusions due to advanced lung cancer.

Adenocarcinoma↗

Recent advances in telepathology.

Telepathology is to transform microscopic and gross images of pathological specimen into digital forms and transmit them electronically to remote sites for consultation and collaboration. Many benefits can be derived from telepathology. Although still at its initial developing stage, this technology is beginning to change the way pathological practice is conducted. With the rapid advancement of computer and telecommunication sciences, telepathology is increasingly becoming more feasible and practical for pathologists. This review gives a comprehensive account of the recent development in telepathology, discusses its functionality, applications, limitations and potential, and serves as an introduction to this rapidly evolving field.

Computer Communication Networks↗