EM chromosome mapping using surface-spread polytene chromosomes

Electron micrographs as well as light micrographs of individual surface-spread polytene (SSP) chromosomes indicate more detailed banding patterns than standard squash preparations do. For EM preparations of SSP chromosomes a simple technique is described, avoiding thin-sectioning of chromosomes.     

Do colonic bacteria contribute to cholesterol gall-stone formation? Effects of lactulose on bile.

Ten healthy middle-aged women volunteered for a study to test the effect of lactulose--a synthetic, non-absorbable disaccharide--on the colonic metabolism of bile acids and on bile lipid composition. Lactulose (60 g daily in eight cases, 39 g daily in two) was taken as a proprietary syrup for six weeks, and bile was collected by duodenal intubation before and immediately after six weeks. All subjects showed a fall in the percentage of the 7-alpha-dehydroxylated bile acid deoxycholic acid (mean 28.4 +/- SEM 3.7 to 15.6 +/- 2.4; p less than 0.002) and a rise in the percentage of the primary bile acid chenodeoxycholic acid (mean 33.2 +/- 42.9 +/- 2.9; p less than 0.001). The percentage of cholic acid rose in eight subjects but mean values did not differ significantly. Bile was initially super-saturated with cholesterol in most subjects and became less saturated with cholesterol in all but one (mean saturation index 1.40 +/- 0.11 to 1.19 +/- 0.07; p less these 0.005). These data support the theory colonic bacteria contribute to cholesterol gall-stone formation.     

Hypophysiotrophic thyrotropin releasing hormone (TRH) synthesizing neurons. Ultrastructure, adrenergic innervation and putative transmitter action

The neuropeptide thyrotropin releasing hormone (TRH) is capable of influencing both neuronal mechanisms in the brain and the activity of the pituitary-thyroid endocrine axis. By the use of immunocytochemical techniques, first the ultrastructural features of TRH-immunoreactive (IR) perikarya and neuronal processes were studied, and then the relationship between TRH-IR neuronal elements and dopamine-beta-hydroxylase (DBH) or phenylethanolamine-N-methyltransferase (PNMT)-IR catecholaminergic axons was analyzed in the parvocellular subnuclei of the hypothalamic paraventricular nucleus (PVN). In control animals, only TRH-IR axons were detected and some of them seemed to follow the contour of immunonegative neurons. Colchicine treatment resulted in the appearance of TRH-IR material in parvocellular neurons of the PVN. At the ultrastructural level, immunolabel was associated with rough endoplasmic reticulum, free ribosomes and neurosecretory granules. Non-labelled axons formed synaptic specializations with both dendrites and perikarya of the TRH-synthesizing neurons. TRH-IR axons located in the parvocellular units of the PVN exhibited numerous intensely labelled dense-core and fewer small electron lucent vesicles. These axons were frequently observed to terminate on parvocellular neurons, forming both bouton- and en passant-type connections. The simultaneous light microscopic localization of DBH or PNMT-IR axons and TRH-synthesizing neurons demonstrated that catecholaminergic fibers established contacts with the dendrites and cell bodies of TRH-IR neurons. Ultrastructural analysis revealed the formation of asymmetric axo-somatic and axo-dendritic synaptic specializations between PNMT-immunopositive, adrenergic axons and TRH-IR neurons in the periventricular and medial parvocellular subnuclei of the PVN. These morphological data indicate that the hypophysiotrophic, thyrotropin releasing hormone synthesizing neurons of the PVN are directly influenced by the central epinephrine system and that TRH may act as a neurotransmitter or neuromodulator upon other paraventricular neurons.