Cultured circular smooth muscle from the rabbit colon

Summary Although cultured vascular smooth muscle cells have been extensively characterized and investigated, there are very few studies of cultured intestinal smooth muscle cells. The aim of this study was to culture colonic smooth muscle (CSM) cells from the rabbit colon. Freshly isolated CSM cells from the circular muscle layer of the distal colon were prepared by collagenase digestion. In primary culture, CSM cells attached to the culture vessels by 48 to 72 h, proliferated by 3 to 7 d, and reached confluency by 14 to 17 d with a “hill-and-valley” pattern. Spontaneous contractions were not observed at any time at 21° or 37° C. Confluent primary cultures were greater than 95% CSM cells, as identified by intensely positive immunofluorescent staining to smooth muscle actin-specific CGA7 and muscle-specific HHF-35 monoclonal antibodies. Transmission electron microscopy of freshly isolated and proliferating CSM cells revealed ultrastructural features consistent with smooth muscle cells. We successfully cultured CSM cells of the rabbit from freshly isolated cells and validated these CSM cells by electron microscopy and immunocytochemical staining. These highly pure primary cultures may be used to investigate numerous aspects of CSM cell metabolism and physiology. These studies were supported by the National Institutes of Health grant to the Inflammatory Bowel Disease Center (Bethesda, MD) P30-AM-32200 and R01-DK-31147. Dr. Kao is the recipient of a Research Career Development Award from the National Foundation for Ileitis and Colitis, Inc. A preliminary report of this work was presented at the American Motility Society Meeting, Houston, TX, in October 1986, and appeared in abstract form inGastroenterology 91: 1057; 1986.     

The presence or absence of light during flotation restricted environmental stimulation: Effects on plasma cortisol,blood pressure,and mood

This study examined the effect of light on relaxation associated with flotation restricted environmental stimulation therapy (REST), as measured by plasma cortisol, mean arterial pressure, and psychometric parameters. Twenty-one subjects were paired by baseline cortisol levels into two groups: one experiencing flotation REST in the presence of light (REST-L) and one experiencing flotation REST in the absence of light (REST-D). Subjects were 15 male and 6 female students aged 22–28 in normal health who had not experienced REST. Repeated flotation REST (8 sessions) either with light or without light was associated with a decrease in plasma cortisol and a decrease in mean arterial pressure, with no differences in effectiveness between groups. The psychometric assessment of mood, using the POMS scale, before and after sessions 1 and 8 revealed mood state improvement in both REST-L and REST-D groups. These data suggest that the presence of light did not compromise the flotation REST experience, as evidenced by the lack of difference between REST-L and REST-D groups.     

Monoploids in Zea mays L. following crosses with untreated and X-rayed pollen

Monoploids in Zea mays L. occur spontaneously among individual diploid seedlings. Plants with the gametic chromosome number have also been detected among members of multiple seedlings of maize and numerous other species of angiosperms. Previous reports disclosed that Xirradiation of the pollen successfully stimulated reduced parthenogenesis in some other angiosperms, but the results of X-ray treatment were inconclusive in maize. Therefore, a tester stock of maize homozygous for lg _land gl _l was crossed with pollen from inbred CI3A, carrying the dominant alleles. The pollen was exposed to 0, 1000, 2000, and 4000r units of X rays. chromosome counts were made from root tips of plants exhibiting both recessive phenotypes to establish the frequencies of monoploids in the control and X₁ populations.Monoploids were more abundant among the individual seedlings from crosses with untreated pollen than in the X₁ populations. X irradiation of the pollen is not a feasible method for the induction of monoploids in maize. The X-ray treatments greatly increased the frequency of multiple seedlings, and deficiencies were numerous among them. The members of a set of multiple seedlings were always genetically identical, and no monoploid members occurred. It is concluded that the induced deficiencies caused atypical development resulting in zygotic or embryonic cleavage.Scientific Article No. A2070, Contribution No. 5023 of the Maryland Agricultural Experiment Station, Department of Botany.     

Methyl bromide as a microbicidal fumigant for tree nuts.

Methyl bromide (MeBr) has broad microbicidal activity, but its use as a disinfectant for food is limited by the resulting bromide residues. Increasing the MeBr concentration, exposure temperature, or exposure period of a treatment tended to increase both the microbicidal efficacy of MeBr and the bromide residues. Its sporicidal activity was less at high than at low relative humidity within the range of 20 to 99%. Both the efficacy and the resulting residues of a MeBr treatment varied inversely with the load of product in a fumigation chamber due to sorption of the fumigant. Fumigation tests with almond kernels inoculated with Escherichia coli or Salmonella typhimurium indicated that MeBr can be used to disinfect whole nut kernels without resulting in excessive bromide residues, although the MeBr level necessary is higher than that normally used for insect control.     

Effect of lithium on granulopoiesis in culture.

Lithium carbonate therapy is associated with polymorphonuclear leukocytosis. In vitro studies have shown that lithium ions stimulate formation of granulocytic colonies. In a study undertaken to determine how lithium acts, colony-forming cells uncontaminated by monocytes (which elaborate colony-stimulating factor [CSF] in vitro) were obtained by means of a two-step cell separation procedure. The effects of lithium on colony formation were then studied in (a) cultures stimulated by humoral CSF, (b) cultures in which monocytes were relied upon to synthesize CSF de novo and (c) unstimulated cultures. Lithium enhanced the action of CSF but did not stimulate colony formation in the absence of CSF. In monocyte-stimulated cultures, colony formation increased with lithium concentrations up to 1 mmol/L but this increase paralleled that in CSF-stimulated cultures and therefore was not due to increased CSF production by monocytes. At higher concentrations of lithium, colony formation decreased in the monocyte-stimulated cultures but increased in the CSF-stimulated cultures. A lithium concentration of 4 mmol/L gave the greatest enhancing effect on colony formation in CSF-stimulated cultures and a concentration greater than 1 mmol/L inhibited de novo synthesis of CSF by monocytes.