Evidence of melanogenesis in porcine retinal pigment epithelial cells in vitro

Several weeks after porcine retinal pigment epithelial (RPE) cell cultures attain confluence, macroscopically visible brown foci appear. The cuboidal cells that form the foci contain numerous phase dark granules that do not exhibit the autofluorescence characteristic of lipofuscin. The data described here indicate that the granules are melanosomes. Electron microscopy revealed three types of electron-dense granules in these cells: simple spheres 0.3-0.5 microns in diameter, large spheres 1-2 microns in diameter, and lysosomal aggregations of the smaller spheres. The matrix of both spheres is composed of 40-nm microvesicles that were also found free in the cytoplasm and aggregated within vacuolar structures. Reversed-phase high-performance liquid chromatography of RPE cells and their media detected melanogens, i.e. intermediates of melanin biosynthesis, including several indole derivatives. The porcine RPE cultures therefore may be a useful system for studying melanogenic regulation.     

NADPH-Diaphorase-Positive Neurons in Primate Cerebral Cortex Colocalize with GABA and Calcium-Binding Proteins

Neurons in the monkey cerebral cortex containing nicotinamideadenine dinucleotide phosphate-diaphorase (NADPH-d) can he dividedinto two distinct types, both nonpyramidal. Type I neurons havea large soma (diameter 20–50 µm), a dense NADPH-dhistochemical reaction. and are distributed throughout the cortex,but mainly in the subcortical white matter, and are mostly aspiny.Type II cells have a small soma (< 20 µm) with lightNADPH-d reactivity and are distributed primarily in the supragranularlayers, particularly layers II and upper III. The numericaldensity of type II cells is much greater than that of type I.Type I neurons also stain for GABA and a few intracortical typeI cells contain calbindin. All type II cells found here arecolocalized with both GABA and calbindin. Neither type I nortype II cells are stained for parvalbumin. Together with previous observations that almost all corticalNADPH-d cells in various subprimates are like type I cells,we suggest that type II cells may form a group of NADPH-d-richneurons differentiated in higher mammalian cortex from a subpopulationof calbindin-containing GABAergic interneurons, and these nitricoxide-synthesizing cells may play a role in control of intracorticalneuronal activity characteristic of higher cerebral functionsin advanced mammals.     

Principles for managing penetrating craniocerebral injuries caused by firearm missiles

Penetrating craniocerebral firearm injuries remain one of the most lethal causes of all trauma and are common both in war or peace time. Data were reviewed for 4140 severely head-injured patients (Glasgow Coma Scale (GCS) scores 3-8) treated at Xi-Jing Hospital between 1973 and 1993; 51 of these patients had acute penetrating craniocerebral injuries caused by firearm missiles. These patients consisted of 46 males (90.2%) and 5 females (9.8%) ranging in age from 3 months to 48 years (median 22.4 years). The lesion types included 2 tangential wounds, 37 tubular wounds and 12 through-and-through wounds. All cases were urgent with the patients in severe and unstable states. After emergency treatment and operation, 5 cases died (9.8%). Follow up studies at three months showed that 23 cases (45.1%) had made a good recovery. Moderate disability, severe disability and vegetative states in this series were 29.4%, 13.7% and 2.0% respectively. Long term follow up studies indicated that 32 were able to resume their occupation. The principles for managing penetrating craniocerebral firearm injuries and suggestions for operation are discussed.     

Influence of pancreas transplantation on cardiorespiratory reflexes, nerve conduction, and mortality in diabetes mellitus

Cardiorespiratory reflexes (CRR) were studied by measuring heart-rate variation during 6 breaths/min respiration and a Valsalva maneuver in 232 insulin-dependent diabetic subjects. Abnormalities were found in 175 patients. During a 7-yr follow-up, 41 (23.4%) patients with abnormal and 2 (3.5%) with normal CRR tests died. The mortality rates of diabetic patients with abnormal autonomic function tests were 17% at 2.5 yr, 33% at 5 yr, and 40% at 7 yr, significantly higher (P less than 0.002) than in patients with normal tests (rates of 4.6, 4.6, and 13.8% at the respective intervals). Nerve conduction studies (NCS) were indicative of somatic neuropathy in 148 of 205 patients. Mortality rates were higher in patients with abnormal NCS than in those with normal results (P less than 0.025). Among patients with abnormal autonomic function, patients with a functioning pancreas transplantation (PTx) had better survival rates than patients with a failed PTx (P less than 0.005) and, on long-term follow-up, better rates than patients without PTx. Similar results were found comparing the same group of patients who had abnormal NCS.     

Effects of an anti-interleuktn-6 (IL-6) murine monoclonal antibody in a patient with acute monoblastic leukemia

Because IL-6 has been involved in the pathogenesis of acute monoblastic leukemia, we investigated thein vitro anti-proliferative effect and thein vivo anti-tumoral effect of an anti-IL-6 murine monoclonal antibody (mAb) in a patient with MSB type acute leukemia. In the current study, we clearly show the IL-6 dependence of monoblastic cell viability and proliferationin vitro in short-term cultures of malignant cells and the clinical activity of the anti-IL-6 murine mAb. The complete neutralization of IL-6in vivo was associated with a transient but complete disappearance of malignant monoblastic cells in the peripheral blood, with improvement or even normalization of several other biological parameters of disease activity, No immunization against the anti-IL-6 murine mAb was observed.     

Penetration of Schistosoma japonicum cercaria into host skin.

The anterior part of Schistosoma japonicum cercaria is a specialized head organ which can slightly stretch out and retract. There are three different types of large unicellular glands in cercarial body, consisting of one head gland, 2 pairs of pre- and 3 pairs of postacetabular glands. These glands differ in position, gross feature, histochemistry and functions. Both polysaccharase and protease activities are demonstrated in the secretions from these glands. Mode of cercarial penetration is described in detail and the penetration is effected by a combination of lytic secretions and mechanical movements. The schematic representation of the process of cercarial penetration is presented. The dynamic distributions of schistosomula in skin at different time intervals after skin penetration in various mammalian hosts are shown. Some newly transformed schistosomula die while penetrating into the skin of 7 mammalian species and the mortality rate varies with the host species, and that can also be affected by the age of cercariae following emergence from the snail. Some physiological aspects between cercariae and newly transformed schistosomula are compared. In contrast to cercariae, schistosomula are saline-adapted and water-intolerant. They were modified histochemically and antigenically.

     

Preservation of human corneas in organ culture: technical protocol

Using organ-culture preservation human corneas were preserved up to 5 weeks in a modified tissue culture medium at 37 degrees C. The endothelial viability was examined after staining with trypan-blue 0.3%, after determination of endothelial cell density and cell loss and by light microscopy after staining with trypan-blue 0.3% and alizarin red 1%. The biochemical analysis of the medium (pH, glucose, lactate) has allowed the evaluation of three kinds of storage. The influence of time preservation on the endothelial viability and the metabolic conditions are determined. At the end of this study a clinical trial is proposed.