Identification of a brefeldin A-insensitive guanine nucleotide-exchange protein for ADP-ribosylation factor in bovine brain.

ADP-ribosylation factors (ARFs) are approximately 20-kDa guanine nucleotide-binding proteins that participate in vesicular transport in the Golgi and other intracellular compartments and stimulate cholera toxin ADP-ribosyltransferase activity. ARFs are active in the GTP-bound form; hydrolysis of bound GTP to GDP, possibly with the assistance of a GTP hydrolysis (GTPase)-activating protein results in inactivation. Exchange of GDP for GTP and reactivation were shown by other workers to be enhanced by Golgi membranes in a brefeldin A-sensitive reaction, leading to the proposal that the guanine nucleotide-exchange protein (GEP) was a target of brefeldin A.

Insulin-like growth factor-1 in Syrian hamsters: interactions of photoperiod, gonadal steroids, pinealectomy, and continuous melatonin treatment.

Four experiments in Syrian hamsters examined the role and possible interaction of photoperiod, gonadal steroids, and the pineal on circulating levels of insulin-like growth factor-1 (IGF-1). In the first experiment, female hamsters were exposed to long photoperiod (LP; 14:10 LD) or short photoperiod (SP; 8:16 LD); an additional group of SP-exposed females was pinealectomized (PX). SP induced a significant depression in IGF-1 concentrations which PX partially prevented.

Effect of ADP-ribosylation factor amino-terminal deletions on its GTP-dependent stimulation of cholera toxin activity.

It has been proposed that the amino-terminal domain of ADP-ribosylation factor (ARF) is critical for its stimulation of cholera toxin ADP-ribosyltransferase activity. In this study, recombinant ARF1 (rARF1), r delta 13ARF1 (recombinant ARF1 lacking the first 13 amino acids) and rPKA14ARF1 (recombinant ARF1 in which the first 14 amino acids were replaced by the first 7 amino acids of the cAMP-dependent protein kinase catalytic subunit) were used to assess the effect of the amino terminus on the ability of ARF to enhance ADP-ribosylation of agmatine by the cholera toxin A subunit. The GTP-dependent ARF activities of r delta 13ARF1 and rPKA14ARF1 were similar to that of rARF1, whereas the GTP requirement for half-maximal activation of cholera toxin A, was somewhat higher for rARF1 than it was for r delta 13ARF1 and rPKA14ARF1.

Alternative splicing of the guanine nucleotide-binding regulatory protein Go alpha generates four distinct mRNAs.

Go alpha a guanine nucleotide-binding (G) protein abundant in brain and other neural tissues, has been implicated in ion channel regulation. Concerted efforts in several laboratories have revealed multiple Go alpha mRNAs and protein isoforms in different contexts. Go alpha is a single copy gene in mammalian species, although the structure, number and tissue localization of Go alpha mRNAs reported by investigators are inconsistent.

Minimally invasive cancer surgery using focused ultrasound: a pre-clinical, normal tissue study.

Using a focused 1.7 MHz ultrasound field (focal length/transducer diameter ratio of 1.7) and in situ intensities spatially averaged within the half-pressure maximum contour in the range 100-400 W cm-2, ablative lesions have been prescriptively placed singly and in arrays, in the livers and bladder walls of adult female Large White pigs.

Pineal acid phosphatase activity in Syrian hamsters: sex differences and effects of castration and androgen replacement therapy.

Pineal acid phosphatase (ACP) activity was examined in seven experiments involving young intact or castrated male and female Syrian hamsters. Nine-week old female hamsters had a 3-fold (p < 0.001) higher ACP activity in their pineal glands than did males.

Richter's hernia of the splenic flexure: computed tomography appearances.

A case of a clinically occult Richter's hernia of the splenic flexure, through an anterior abdominal wall defect, is described. In view of the initial absence of bowel obstruction with a partial or Richter's hernia, and in the absence of physical findings, the importance of computed tomography (CT) in demonstrating abdominal wall lesions is highlighted.

New immunohistochemical "sandwich" staining method for mdr1 P-glycoprotein detection with JSB-1 monoclonal antibody in formalin-fixed, paraffin-embedded human tissues.

We have developed a new immunoperoxidase "sandwich" staining method for amplified detection of P-glycoprotein (Pgp) that is suitable for use on formalin-fixed, paraffin-embedded (conventional) tissue sections. This was accomplished by substantially changing the procedure described by Chan (1988) so as to increase specific staining intensity and to decrease nonspecific background staining. To determine the most appropriate primary antibody for the assay, we compared the immunoreactivity of JSB-1, C494, and C219 monoclonal antibodies recognizing internal epitopes of Pgp, and MRK16 and 4E3 monoclonal antibodies recognizing external epitopes of Pgp.

Lesion development in focused ultrasound surgery: a general model.

An analytical model has been constructed for the process of formation of thermal lesions in tissue, resulting from exposure to intense, highly focused ultrasound beams such as may be used in minimally invasive surgery. The model assumes a Gaussian approximation to beam shape in the focal region and predicts, for any such focal beam, the time delay to initiation of a lesion and the subsequent time course of growth of that lesion in lateral and axial dimensions, taking into account the effects of thermal diffusion and blood perfusion. The necessary approximations and assumptions of the model are considered.

The contribution of hospitals to a local economy: a case study in Iowa and Illinois.

This study assessed the importance of the hospital sector to a regional economy by examining the estimated effects of direct and indirect hospital spending on the income and employment of a metropolitan region in Iowa and Illinois. The evaluation included the role of hospital services as a regional "export." In addition, the stabilizing impact of hospital spending during business cycles was examined.

Cardiovascular and neuroendocrine responses to extended laboratory challenge.

The purpose of this study was to examine the effects of a 2-hour laboratory challenge on heart rate, blood pressure, catecholamines, and cortisol; and investigate the contribution of the physical act of speaking on both neuroendocrine and cardiovascular measures. Using a within-subjects design, 14 subjects were tested individually during two separate laboratory sessions. During one session, subjects engaged in two cognitively demanding tasks for 2 hours.

Muskrat populations in Virginia's Elizabeth River: physiological condition and accumulation of environmental contaminants.

This study evaluated the physiological condition and environmental contaminant concentrations in muskrats inhabiting the contaminated lower region of the Elizabeth River, Virginia, (USA). Muskrats trapped in the lower region of the Elizabeth River weighed less, had lower mean fat indexes, lower relative spleen weights, greater relative adrenal weights, and an increased incidence of disease and parasitism compared to muskrats trapped in the less contaminated upper region of the Elizabeth River or Nansemond River. Concentrations of the metals aluminum, cadmium, copper, nickel, and zinc and the number of polyaromatic hydrocarbon compounds were greater in lower Elizabeth River muskrats compared to upper region Elizabeth River or Nansemond River muskrats.

Rapid prenatal diagnosis of Patau's syndrome in a fetus with an abdominal wall defect by 72 hour culture of cells from amniotic fluid.

A woman in the 32nd week of pregnancy was referred for investigation because of fetal abnormalities, including an abdominal wall defect, detected by ultrasonography. In view of the increased risk of chromosome abnormality, amniocentesis was performed to enable informed decisions about the management of the pregnancy and delivery to be taken. Cells from the liquor were inoculated into standard lymphocyte culture medium and incubated for 72 h.

Development and androgen regulation of the secretory cell types of the Syrian hamster (Mesocricetus auratus) Harderian gland.

The secretory cell types of the hamster Harderian glands were studied in both male and female Syrian hamsters. As previously demonstrated, female hamsters showed a single secretory cell type (type I), while male hamsters displayed two secretory cell types (type I and type II). Type-II cells were observed after the first month of age correlating with the increase in testosterone levels.

Characterization of recombinant and endogenous ADP-ribosylation factors synthesized in Sf9 insect cells.

ADP-ribosylation factors (ARFs) are a family of highly conserved, 20-kDa guanine nucleotide-binding proteins that participate in protein trafficking and enhance cholera toxin-catalyzed ADP-ribosylation. ARF 2 from bovine retinal cDNA was expressed in Sf9 insect cells using recombinant baculovirus and compared to the major insect cell ARF (Sf9 ARF) and to recombinant ARF 2 expressed in Escherichia coli (E. coli rARF 2).

Molecular characterization of a conserved, guanine nucleotide-dependent ADP-ribosylation factor in Drosophila melanogaster.

ADP-Ribosylation factors (ARFs) are ubiquitous approximately 20-kDa guanine nucleotide-binding proteins that stimulate cholera toxin-catalyzed ADP-ribosylation in vitro. Because the functional role(s) of ARF in mammalian systems is (are) elusive, we looked for ARF in Drosophila melanogaster, and report the partial purification and molecular cloning of an ARF from Drosophila. We cloned the Drosophila ARF 1 gene without library screening by a combination of 5 polymerase chain reactions (PCRs), yielding a 546-base open reading frame encoding 182 amino acids, which are > 93% identical to those of mammalian class I ARFs.

Gender-associated differences in the development of 5-aminolevulinate synthase gene expression in the harderian gland of Syrian hamsters.

The mRNA levels for aminolevulinate synthase (ALV-S), the rate-limiting enzyme in porphyrin synthesis, were studied in male and female Syrian hamsters during postnatal development. Sex-associated differences in the expression of ALV-S gene were evident at the end of the third week of postnatal development. Serum levels of luteinizing hormone (LH), testosterone, cortisol, thyroid hormones and insulin-like growth factor were also studied in order to correlate their concentrations with the mRNA levels for ALV-S.

Effects of brefeldin A and accessory proteins on association of ADP-ribosylation factors 1, 3, and 5 with Golgi.

ADP-ribosylation factors (ARFs) are approximately 20-kDa guanine nucleotide-binding proteins initially identified by their ability to enhance in vitro cholera toxin-catalyzed ADP-ribosylation and subsequently shown to participate in vesicular transport in the Golgi and other cellular compartments. By cDNA and genomic cloning, at least six mammalian ARFs were identified. Brefeldin A (BFA) disrupts Golgi membranes and inhibits binding of soluble high molecular weight proteins to Golgi fractions.

ARD 1, a 64-kDa guanine nucleotide-binding protein with a carboxyl-terminal ADP-ribosylation factor domain.

Clones referred to as ARD 1 were isolated from human and rat cDNA libraries. ARD 1 genes encode a putative 64-kDa protein that contains an 18-kDa ADP-ribosylation factor (ARF) domain at the carboxyl terminus and is much larger than the other monomeric approximately 20-kDa guanine nucleotide-binding ARF proteins thus far identified. ARD 1 mRNAs of 3.

Characterization of the human ADP-ribosylation factor 3 promoter. Transcriptional regulation of a TATA-less promoter.

The 5'-flanking region of the human ADP-ribosylation factor 3 gene contains the features of a housekeeping gene. It lacks a TATA or CAAT box, has several GC boxes within a highly GC-rich region, and utilizes multiple transcription initiation sites. The cis-acting elements involved in regulating expression of the gene were identified by transient transfections of IMR-32 neuroblastoma cells.

Effect of myristoylation on GTP-dependent binding of ADP-ribosylation factor to Golgi.

ADP-ribosylation factors (ARFs), a family of approximately 20-kDa guanine nucleotide-binding proteins that activate cholera toxin ADP-ribosyltransferase in vitro, have been implicated in intracellular protein trafficking and are thought to cycle between cytosolic and membrane compartments. Although isolated predominantly as soluble proteins, ARFs associate with membranes and phospholipids in a GTP-dependent manner. In contrast to other small GTP-binding proteins, ARFs are NH2 terminally myristoylated.