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Biomedical subjects

L Holland

Publications and source records attributed to L Holland.

11 recordsLinked to original sources

Development of invasive and growth factor-independent cell variants from primary human melanomas.

Growth autonomy and high levels of invasiveness are characteristics of human melanoma cells that are metastatic in vivo. By consecutive passage through a reconstructed basement membrane, we have selected from 5 of 6 primary melanoma cell lines variants which show an up to 10-fold increase in invasiveness. The invasive variants grew more rapidly than the parental, noninvasive cells in serum- and growth factor-free medium and one of the 3 variant cell lines with the highest invasive capacity in vitro metastasized to the lungs when injected s.c. into nude mice. In a second approach, variants of 6 primary melanoma cell lines were clonally selected in medium without exogenous growth factors (protein-free medium). These selected cells showed higher invasive properties in vitro and in vivo than the parental cells. Clones of invasive and growth factor-independent cell variants were heterogenous and changed over time in the absence of selected pressure to a phenotype similar to that of parental nonselected cells. These results indicate that primary melanoma cells contain subpopulations of cells that have the phenotype of an advanced (metastatic) stage of tumor progression, but this phenotype is not stable without selective pressure.

Growth Substances

The influence of oral potassium chloride on blood pressure in hypertensive men on a low-sodium diet.

Clinical and epidemiologic studies suggest that the intake of potassium chloride lowers blood pressure. To investigate whether supplemental potassium chloride (96 mmol of microcrystalline potassium chloride a day) reduced the need for antihypertensive medication in hypertensive men on a restricted-sodium diet, we conducted a randomized, placebo-controlled, double-blind clinical trial. A total of 287 men 45 to 68 years of age, 142 given potassium chloride and 145 given placebo, were followed for an average of 2.2 years after the withdrawal of their antihypertensive medication. Men in both groups received instructions on following a low-sodium diet. Overnight urinary sodium excretion fell from 63 mmol per eight hours at base line to an average of 45 mmol per eight hours during follow-up. Participants given supplemental potassium chloride had significantly higher (P less than 0.001) serum potassium levels and urinary potassium excretion (averaging 4.5 mmol per liter and 42.5 mmol per eight hours, respectively) during follow-up than participants given placebo (4.2 mmol per liter and 20.0 mmol per eight hours). Seventy-nine participants in each group required reinstitution of antihypertensive medication according to strict indications defined by the protocol. No significant differences in systolic or diastolic blood pressure were observed between the two groups. During follow-up, systolic and diastolic blood pressure averaged 130.6 and 82.5 mm Hg, respectively, for participants given supplemental potassium, and 132.5 and 83.1 mm Hg for participants given placebo. We conclude that supplemental potassium chloride does not reduce the need for antihypertensive medication in hypertensive men on a restricted-sodium diet.

Administration, Oral

Viral nucleic acid synthesis in HSV infected neural cells.

HSV-1 replication and synthesis of viral DNA and RNA have been examined in gliomas of human (COX) and rat origin (C6) and in mouse neuroblastomas (D2). COX cells fully support HSV-1 replication and show patterns of viral DNA and RNA synthesis similar to those seen in continuous line cells. HSV-1 also grows to high titers in D2 cells but without concomitant high levels of viral DNA and RNA synthesis in the infected cells. Finally, HSV-1 established a persistent infection in C6 cells. Viral mRNA and DNA synthesis could not be detected in these cultures. At cycles of approximately 15--20 days, the persistently infected cultures exhibited massive CPE and relatively high production of infectious HSV.

Animals

Ionic mechanism of the fertilization potential of the marine worm, Urechis caupo (Echiura).

Microelectrode and tracer flux studies of the Urechis egg during fertilization have shown: (a) insemination causes a fertilization potential; the membrane potential rises from an initial level of -33 +/- 6 mV to a peak at +51 +/- 6 mV (n = 16), falls to a plateau of about +30 mV, then returns to the original resting potential 9 +/- 1 min (n - 10) later; (b) the fertilization potential results from an increase in Na+ permeability, which is amplified during the first 15 s by a Ca++ action potential; (c) the maximum amplitude of the fertilization potential, excluding the first 15 s, changes by 51 mV for a 10-fold change in external [Na+]; (d) in the 10 min period after insemination, both Na+ and Ca++ influxes increase relative to unfertilized egg values by factors of 17 +/- 7 (n = 6) and 34 +/- 14 (n = 4), respectively; the absolute magnitude of the Na+ influx is 16 +/- 6 times larger than that of Ca++; (e) in the absence of sperm these same electrical and ionic events are elicited by trypsin; thus, the ion channels responsible must preexist in the unfertilized egg membrane; (f) increased Na+ influx under conditions of experimentally induced polyspermy indicates that during normal monospermic fertilization, only a fraction of available Na+ channels are opened; we conclude that these channels are sperm-gated; (g) Ca++ influx at fertilization is primarily via the membrane potential-gated channel, because kinetics are appropriate, and influx depends on potential in solutions of varying [Na+], but is independent of number of sperm incorporations in normal sea water.

Action Potentials

Effects of respiration on cardiac performance.

The conventional explanation for the fall in left ventricular stroke volume (LVSV) with inspiration is that blood pools in the lungs, thereby decreasing pulmonary venous return. In anesthetized dogs, we have found an increase in left ventricular filling pressure (LVFP) with both constant and increasing lung volume during an inspiratory effort. Transmural aortic diastolic pressure rises as LVSV falls and LVFP rises consistent with the hypothesis that a fall in pleural pressure afterloads the left ventricle. Additionally the increase found in right ventricular filling pressure with inspiration may adversely affect LV performance by decreasing LV compliance and/or contractility. Our findings are incompatible with pooling of blood in the lungs being the primary determinant of the fall in LVSV with inspiration.

Animals

Bleeding from self-administration of phenindione: a detailed case study.

A young woman presented with a 2 year history of a severe bleeding disorder and marked deficiencies in all four vitamin-K-dependent factors. Metabolic studies with tracer doses of tritium-labelled vitamin K1 suggested that the patient might be taking an oral anticoagulant; and subsequently her plasma was found to contain a substance identical to phenindione in its spectrophotometric and chromatographic properties. The half-disappearance times of factors II, IX, X were measured after the administration of a concentrate of these factors and were found to conform with published figures. The concentrate controlled the patient's excessive bruising and prolonged skin and gingival bleeding. It would therefore seem that factor VII may not be essential in reversal of the bleeding disorder induced by anticoagulant overdose.

Adult

Oocyte differentiation in Urechis caupo (Echiura): a fine structural study.

The fine structure of oocytes of Urechis caupo is described for seven arbitrary stages ranging from the smallest oocytes (7 mum in diameter) in the coelom to the mature oocytes (115 mum in diameter) in the storage organs. Although most types of cytoplasmic organelles accumulate more or less continuously, yolk granules do not appear until oocytes reach a diameter of 35 mum, and there is stage-specific synthesis of cortical granules in 60-80 mum oocytes. In the nucleus a single nucleolus first appears when an oocyte is 15 mum in diameter. Then a nucleolus satellite, which is about 3 mum in diameter, forms in 30 mum oocytes; this nucleolus satellite later (60-70 mum oocytes) becomes surrounded by 750 nm dense spherical bodies. Large (2-4 mum in diameter) juxtachromosomal spherules occur only in the nuclei of mature oocytes. Microvilli become progressively more numerous and longer until the oocyte reaches a diameter of 90 mum; their tips project 1 mum beyond the fibrous surface coat, which is 2 mum thick when well developed. Near the end of oocyte growth, the microvilli retract into the surface coat leaving their pinched-off tips adhering to the outside of the coat.

Animals

Optimal leukocyte removal from refrigerated blood with the IBM 2991 blood cell processor.

One hundred forty-six units of two to five-day-old refrigerated blood were washed with the IBM Blood Cell Processor. The ABO and Rh types of the units varied according to available inventory. This study was designed to select the best protocol from the 42 possible combinations utilizing the machine's variable red blood cell override (RCO). A one liter wash program was used. The two parameters selected for study were the per cent red blood cell recovery and the per cent white blood cell removal. The protocol of choice uses a RCO of four seconds in the first and second steps of the program. This protocol yields a washed unit with a mean red blood cell recovery of 82 per cent and a mean white cell removal of 93 per cent. Our studies indicate that the IBM Blood Cell Processor can be used to wash two to five day old refrigerated blood for the preparation of leukocyte-poor blood that would meet the Standards of the AABB.

Blood Preservation