Search PubMed⌕ Search

Biomedical subjects

G J Hausman

Publications and source records attributed to G J Hausman.

At least 19 recordsLinked to original sources
PubMed1992

Alterations of serum insulin-like growth factor-I (IGF-I) and IGF-binding proteins (IGFBPs) in swine infected with the protozoan parasite Sarcocystis miescheriana.

The effects of a Sarcocystis miescheriana infection on insulin-like growth factor-I (IGF-I) and insulin-like growth factor binding proteins (IGFBPs) were investigated to determine possible mechanisms of growth retardation in growing pigs. Sixteen pigs averaging 14 kg body weight were divided into 4 groups of 4 pigs each and infected either with 0.5, 1.0, or 3.0 x 10(6) sporocysts of S. miescheriana. Four pigs were retained as non-infected controls; however, they became serologically positive during the course of the infection. Effects also were investigated in 2 groups of 3 pregnant sows. One group was infected with 0.5 x 10(6) sporocysts and the other group was retained as uninfected controls. Body weights of infected growing pigs were depressed as compared to controls following the acute phase 15 d after infection (dai). Serum concentrations of IGF-I dropped significantly (p < 0.05) during the acute phase of infection in all infected groups of growing pigs. Conversely, the amounts of unsaturated serum IGFBPs were elevated significantly (p < 0.05) during the acute phase of infection. Specifically, serum concentrations of IGFBP-1, IGFBP-2, and IGFBP-4 were elevated at this time, as determined by ligand blot analysis. There was no association between growth factor alterations and tissue damage as measured by serum creatinine kinase and aspartate aminotransferase levels. The extent of effects in growing pigs was related to the amount of the original parasite inoculum. During the acute phase of infection 2 of 3 pregnant sows aborted. The third sow went to term, but piglets were stillborn or died within 24 hr. Compared to uninfected controls, serum concentrations of IGF-I in infected pregnant sows were depressed during and after the acute phase of the infection. Levels of unsaturated serum IGFBPs in pregnant sows were not affected. These data suggest that decreased IGF-I levels and/or elevated levels of specific forms of IGFBPs may be a mechanism by which growth is affected in feeder pigs infected with S. miescheriana.

Animals↗
PubMed1992

Biochemical and cytochemical studies of preadipocyte differentiation in serum-free culture of porcine stromal-vascular cells: interaction of dexamethasone and growth hormone.

Stromal-vascular cells from adipose tissue of pigs 5-7 days of age were grown in serum for 2-3 days and switched to serum-free (insulin, transferrin and selenium) conditions +/- test hormones for 6-7 days. The interaction of dexamethasone (DEX) and human growth hormone (hGH) was evaluated since glucocorticoids augment and hGH antagonizes the effect of insulin. Low levels (1-10 nM) of DEX with insulin doubled (p less than 0.05) specific activity of glycerol phosphate dehydrogenase (GPDH) and doubled (p less than 0.05) the number of detectable fat cells relative to insulin alone. DEX with insulin enhanced the morphological differentiation of preadipocytes and markedly increased fat cell cluster numbers in the presence of hGH. Furthermore, 1-10 nM of DEX partially blocked (p greater than 0.05) the inhibitory effect of 10 nM hGH on GPDH activity, but 1-100 nM DEX had no effect (p greater than 0.05) on the ability of hGH to compromise lipid deposition. DEX alone (no insulin or hGH) induced the appearance of esterase-reactive but lipid-free cells. Cells with these characteristics were increased in number by DEX in the presence of hGH but were nearly absent in the presence of insulin and DEX. Therefore, transient exposure to GH in vivo may have no permanent effect on adipose tissue development in the continued presence of glucocorticoids.

Adipose Tissue↗
PubMed1992

The influence of thyroxine on the differentiation of adipose tissue and skin during fetal development.

Preobese fetuses have elevated thyroid hormone levels and depressed growth hormone levels relative to lean fetuses. Therefore, we are studying various experimental fetal pig models to explore the relationship between endocrine status and onset of obesity. In the present study, intact and hypophysectomized (d 70) fetuses were implanted with thyroxine (T4) pellets on d 70 of gestation, and blood, adipose tissue, and skin samples were obtained upon removal of d 90 of gestation. Body weights were similar for all groups and T4 treatment reversed myxedema in hypophysectomized fetuses. Serum T4 levels were elevated (p less than 0.05) and skin and hair development were enhanced (p less than 0.05) to a similar degree by T4 treatment in intact and hypophysectomized fetuses. However, T4 did not influence adipose tissue development in intact fetuses, but markedly enhanced development in hypophysectomized fetuses. For instance, fat cell size and lipogenic enzyme activities in hypophysectomized fetuses were increased (p less than 0.05) by 5-mg and 15-mg T4 treatments, with a marked increase (p less than 0.05) in apparent fat cell number with the 15-mg T4 treatment. In contrast, there was no effect of T4 (15 mg) on these parameters in intact fetuses. Therefore, fetal obesity may be directly associated with elevated thyroid hormone levels and suppressed growth hormone levels, but not with elevated T4 levels alone.

Adipose Tissue↗
PubMed1992

Responsiveness to adipogenic agents in stromal-vascular cultures derived from lean and preobese pig fetuses: an ontogeny study.

Primary cultures of stromal-vascular (S-V) cells from adipose tissue were used to evaluate characteristics of preadipocytes from lean and preobese fetuses at several ages (50, 75, and 110 d). In insulin-supplemented (1 microM) cultures (serum free) there was a significant age x fetal genotype interaction (P less than .01) for glycerol-phosphate dehydrogenase specific activity (GPDH); GPDH activity was genotype-dependent at 110 d (preobese greater than lean). The responses of S-V cultures (preadipocyte development) to 2% pig serum and to insulin (serum free) were similar. Main effects of genotype and age were significant (P less than .05) for protein levels in pig serum and insulin-treated cultures. There was a significant genotype x age (P less than .05) interaction for GPDH activity and protein levels in cultures treated with dexamethasone + 3-isobutyl-1-methylxanthine (DEX-IBMX). Treatment with DEX-IBMX induced more preadipocyte development in cultures from preobese fetuses than in cultures from lean fetuses at 110 d (P less than .05). The responsiveness of S-V cultures to DEX-IBMX (enhanced development) increased considerably between 50 and 75 d regardless of fetal genotype, but there was little response in cultures form 50-d fetuses. Preadipocyte development in lean and preobese fetuses diverged between 75 and 110 d, resulting in many more preadipocytes in preobese fetuses at 110 d. Therefore, S-V cells from preobese fetuses (late term) may be inherently more sensitive to adipogenic agents than S-V cells from lean fetuses.

1-Methyl-3-isobutylxanthine↗
PubMed1992

Norepinephrine inhibits rat pre-adipocyte proliferation.

Hormonal and neural status are major determinants for cellular growth. The purpose of this study was to assess the influence of the adrenergic hormones on pre-adipocyte growth in primary cell culture. Stromal-vascular cells were obtained from the inguinal pad of young rats and grown in culture for two weeks. Cells were exposed to norepinephrine (NE) during the proliferative phase of growth, labelled by [3H]-thymidine incorporation and then placed on a differentiation promoting medium. Adipocytes and stromal cells were separated using a density gradient, and [3H]-thymidine content was determined for both cell types. NE reduced [3H]-thymidine uptake indicating a reduction in pre-adipocyte proliferation. NE-induced inhibition of pre-adipocyte growth was blocked by the presence of propranolol, whereas phenoxybenzamine had no effect, thereby suggesting that NE-inhibition is through beta-adrenoceptors. Pre-adipocytes were treated with NE for varying lengths of time to investigate whether cells were desensitized to chronic beta-adrenergic stimulation. In addition, adenosine deaminase (ADA) was also applied to eliminate adenosine which may accumulate during NE stimulation. Neither the duration of NE exposure nor ADA treatment affected adrenergic control of adipocyte growth. These studies indicate that NE reduces pre-adipocyte proliferation and therefore may be an important negative regulatory component of adipocyte growth.

Adenosine Deaminase↗
PubMed1992

Expression of transforming growth factor-beta (TGF-beta 1) and insulin-like growth factor II (IGF-II) messenger RNA in the developing subcutaneous tissue (SQ) of the fetal pig.

Studies (in situ hybridization) were performed on the developing subcutaneous tissue (SQ) of the pig fetus to determine sites of synthesis of TGF-beta 1 and IGF-II. Tissues from 50, 70, 90, 110-day-old pig fetuses and 7-day-old postnatal pigs were Bouin's fixed, paraffin embedded and hybridized with biotin labelled cDNA probes. The expression of TGF-beta 1 messenger RNA was detectable primarily in the dermis, hair follicle fat lobule, outer and inner SQ areas at all ages. Cells adjacent to adipocyte clusters and some small adipocytes were positive for TGF-beta 1. There was no positive hybridization of TGF-beta 1 probes in the developing muscle below the SQ. The expression of IGF-II was evident in the developing muscle below the SQ at 50 d and 70 d, and could be detected in the outer and inner SQ at 70 d. Much lower levels of IGF-II expression were observed in 90 and 110 d fetuses, but an increase in IGF-II expression was evident in the muscle of 7d postnatal pigs. Our results suggest that paracrine and autocrine regulatory systems may be operative for developing adipocytes (TGF-beta 1) and muscle (IGF-II), with specific areas and times of TGF-beta 1 and IGF-II expression being evident in developing pig SQ tissue.

Animals↗
PubMed1991

Cytoplasmic proteins of porcine adipocytes: identification with monoclonal antibodies.

In the present study, monoclonal antibodies were produced using porcine adipocyte extracts as the immunogen. Two of the monoclonal antibodies, designated CB6 and IB4, exhibited reactivity toward only cells containing lipid in stromal-vascular cell cultures. The antigens recognized by the CB6 and IB4 monoclonal antibodies were 50 kD and 55 kD proteins, respectively. In vivo, IB4 immunoreactivity was detected only in lipid-containing cells, whereas immunofluorescence using CB6 was also detectable around muscle fiber bundles underlying the subcutaneous mesenchyme. In fetal subcutaneous mesenchyme, CB6 and IB4 immunoreactivities toward lipid-containing cells increased with developmental age, but each was not detectable in cells containing the smallest lipid droplets. In stromal-vascular cultures containing adipocytes, 48 hour treatment with the anti-lipogenic agent, growth hormone, only slightly altered CB6 immunoreactivity, whereas IB4 immunoreactivity was reduced by more than sixfold. The exact identity of the CB6 and IB4 antigens was not determined, but each may be useful as markers for studying regulation of adipocyte metabolism.

Adipose Tissue↗
PubMed1991

Vascular and cellular development in fetal adipose tissue: lectin binding studies and immunocytochemistry for laminin and type IV collagen.

A cytochemical study of vascular and cellular development in fetal adipose tissue was conducted utilizing 10 plant lectins (fluorescein isothiocyanate (FITC)-labeled), antibodies against laminin, types II and IV collagen, and a probe for actin. Throughout fetal development (50-110 days) blood vessels were stained by galactose binding lectins and stained for actin, type IV collagen, and laminin. Adipocyte reactivity for laminin was strong throughout development, whereas adipocyte staining for type IV collagen and several lectins increased from weak to moderate between 70 and 110 days of fetal life. In general, staining intensity for lectins was greater for blood vessels than for adipocytes at every age, and staining for lectins and type IV collagen was detected much earlier on blood vessels than on adipocytes. However, the ontogeny and intensity of laminin staining were similar for developing adipocytes and vasculature. Adipocyte staining by several lectins was dependent on location within the tissue, whereas blood vessel lectin staining was not location-dependent. Neuraminidase pretreatment abolished the variation in cellular lectin staining due to location (within the tissue) but did not alter age-related changes in cellular staining. This study indicates that the differentiation of the extracellular matrix of blood vessels and adipocytes is clearly distinct in regard to glycoconjugate composition and temporal pattern of glycoconjugate and type IV collagen deposition.

Actins↗
PubMed1991

Concentration of insulin-like growth factors (IGF-I and IGF-II) in tissues of developing lean and obese pig fetuses.

An ontogeny study of lean and pre-obese pig fetuses was conducted to evaluate the temporal and genetic influence on the relationship between growth and IGF production. At 70, 90 and 110 days of gestation, three fetal pigs were obtained from each of three obese and lean dams. The heart, kidneys, lungs, liver and one biceps femoris muscle were removed, weighed, frozen and extracts prepared. For all the organ and tissue weights, there were significant (P less than .001) main effects of genotype (G) and fetal age (A). At each fetal age, pre-obese fetuses had lighter organ and tissue weights. The content of IGF's in serum and various tissues and/or organs was also determined. Main effects of A and G were significant (P less than .05) for liver and lung IGF-I and muscle IGF-II concentrations. Main effects of G were significant (P less than .01) for liver IGF-II whereas muscle IGF-I and lung IGF-II concentrations were only affected by fetal age (P less than .01). Overall, liver IGF-I and II and muscle IGF-II concentrations were higher in lean fetuses (P less than .01). Serum IGF-I concentration increased (P less than .01) with fetal age and was independent of fetal strain. Serum IGF-II concentration increased with age (P less than .01) in lean but not pre-obese fetuses. Therefore, these studies demonstrate genotype dependent growth factor concentrations in tissues and organs of developing fetuses. Finally, serum concentration of IGF's were strongly correlated (positive) with overall growth but were not strongly correlated with tissue and organ concentration of IGFs.

Animals↗
PubMed1991

Metabolic development of liver and adipose tissue in pre-obese and control pig fetuses.

The metabolic development of the liver and adipose tissue was examined in 75 and 110 day fetuses from genetically obese and control sows. In the liver, glucose and palmitate utilization was influenced by both age and strain. Higher rates of glucose oxidation and palmitate oxidation and esterification were observed in the 75 day compared to the 110 day fetuses. Hepatic palmitate oxidation was greater in pre-obese than in control fetuses at both fetal ages, while hepatic palmitate esterification was greater in pre-obese than in control fetuses at 75 days of gestation only. In subcutaneous adipose tissue, de novo lipogenesis increased with age and was higher in pre-obese than in control fetuses by 110 days of gestation. At 75 days of gestation, glucose oxidation and incorporation into fatty acids was similar in adipose tissue from both strains. However, by 110 days of gestation, both basal and insulin-stimulated rates of glucose metabolism were greater in pre-obese compared to control fetuses. Palmitate esterification increased with age but was similar in pre-obese and control fetuses. Basal lipolysis was not affected by strain or age. Isoproterenol had no effect on lipolysis in the 75 day fetuses while stimulating glycerol release to a comparable degree in 110 day fetuses of both strains. This study demonstrates that metabolic differences between genetically obese and control pigs are already apparent in the pre-obese state prior to birth. Such alterations in hepatic and adipose tissue carbohydrate and lipid metabolism, which promote early lipid storage by the pre-obese fetuses, may serve as useful metabolic markers for the development of obesity.

Adipose Tissue↗
PubMed1990

Decreases in local hormone biosynthesis and c-fos gene expression accompany differentiation of porcine preadipocytes.

To better understand possible autocrine or paracrine mechanisms involved in adipose tissue development, we have studied the biosynthesis of insulinlike growth factor I (IGF-I) and prostaglandin E2 (PGE2) by cultured porcine preadipocytes in response to factors known to modulate cell growth and differentiation. The expression of c-fos was also monitored because of the potential role of that proto-oncogene in coordination of growth and differentiation. Preadipocytes were grown to confluence and then maintained in one of three media treatments: a) standard medium supplemented with 10% fetal bovine serum (FBS), b) FBS supplemented with dexamethasone (Dex), c) FBS supplemented with dibutyryladenosine 3'-5'-cyclic monophosphate. Indirect measurements of growth indicated that cell proliferation did not differ due to media type. Histochemical and enzymatic measurements of adipocyte development revealed that differentiation occurred only in those cultures exposed to Dex. The increase in adipocyte differentiation in response to Dex was associated with a decrease in c-fos and actin RNA expression whereas the decrease in c-fos RNA expression in response to Dex was small (approximately 40%); immunocytochemical analysis indicated that induction of Fos protein occurred only in undifferentiated cells. Thus, the cells responsible for the decrease in c-fos RNA expression are possibly those signaled to differentiate into adipocytes. Expression of IGF-I RNA and secretion of IGF-I and PGE2 were also decreased in response to Dex treatment. These data provide the first demonstration that biosynthesis of IGF-I by preadipocytes can be modulated by a potent inducer of adipocyte differentiation. The combined results indicate that glucocorticoids may stimulate adipocyte differentiation by suppressing intracellular and putative intercellular mitogenic signals.

Actins↗
PubMed1990

Regulation of insulin-like growth factor-I ribonucleic acid expression, polypeptide secretion, and binding protein activity by growth hormone in porcine preadipocyte cultures.

Insulin-like growth factor-I (IGF-I) is a mitogenic polypeptide postulated to mediate the effect of GH on adipose tissue development. To determine if the effect of GH could be mediated by the local production of IGF-I, we have characterized IGF-I RNA expression, polypeptide secretion, and binding protein activity in primary preadipocyte cultures derived from porcine adipose tissues. GH acutely regulated the abundance of multiple IGF-I RNA transcripts and resulted in a 2-fold increase in secreted immunoreactive IGF-I (iIGF-I) polypeptide in medium conditioned for 48 h by preadipocyte cultures relative to those not receiving GH. Immunocytochemical data indicated that IGF-I is synthesized by presumptive and mature adipocytes. The effect of GH on iIGF-I secretion was observed in cultures derived from both fetal and postnatal animals, while secreted IGF-binding protein activity was increased due to GH only in cultures from fetal animals. The increase in local IGF-I production in response to GH was associated with a decrease in adipocyte development, suggesting that local IGF-I may contribute to suppression of differentiated phenotype.

Adipose Tissue↗
PubMed1990

Adipose tissue development in the fetal pig examined using monoclonal antibodies.

Two anti-adipocyte monoclonal antibodies (MAbs: AD-1 and AD-2) have been used to study the development of dorsal s.c. adipose tissue in fetuses from 50 to 110 d of gestation. Immunofluorescent staining of cryostat sections with each antibody revealed antigen-positive cells in fetal s.c. mesenchyme prior to lipid deposition. Lipid droplets as well as AD-1 and AD-2 positive cells were detected within the underlying muscle at 50 d. From 70 to 110 d of development, the AD-1 and AD-2 MAbs each detected all adipocytes examined, as well as capillaries associated with fat cell clusters in s.c. tissues. Reactivity toward both antibodies, as well as lipid deposition, also was detectable in the muscle underlying the s.c. mesenchyme from 70 d onward. Each MAb possessed a distinct pattern of reactivity. The AD-2 MAb stained arrector pili muscles and vessels in the s.c. mesenchyme and vessels in the underlying muscles, whereas the AD-1 did not. No reactivity using either MAb was detectable toward any other cell types within s.c. tissues. These results established the presence of cells expressing surface determinants found on mature adipocytes and associated capillaries prior to adipogenesis. A lineage relationship between adipocytes and capillary endothelial cells is suggested.

Adipose Tissue↗
PubMed1990

In vivo studies of lipid metabolism in lean and pre-obese swine.

Selection for high and low backfat in swine has resulted in obese and lean strains of swine. Fetuses (110 days) from obese dams (HF fetuses) when compared to fetuses from lean dams (LF fetuses) have larger fat cells, higher rates of palmitate esterification in adipose tissue and much higher levels of adipose tissue lipoprotein lipase (LPL) activity. In the present study an exogenous lipid load was given to HF and LF fetuses; lipid uptake into adipose tissue and liver was quantified as was the clearance of lipid from the blood. Uptake of lipid by liver was significant and similar for HF and LF fetuses. However, adipose tissue from HF fetuses took up significantly more lipid than did adipose tissue from LF features. Likewise, lipid clearance was greater in HF fetuses than in LF fetuses. Therefore, the amount of LPL activity in fetal adipose tissue is indicative of the functional capacity of the adipocytes to take up lipid and remove it from the blood.

Adipose Tissue↗
PubMed1990

Monoclonal antibodies against cell surface antigens expressed during porcine adipocyte differentiation.

Two monoclonal antibodies (AD-1 and AD-2) were prepared by fusion of mouse myeloma cells and lymph node cells of mice immunized with porcine adipocyte plasma membranes. Immunoprecipitation of iodinated adipocyte plasma membrane proteins followed by SDS-PAGE and autoradiography yielded protein antigens for each antibody. The AD-1 and AD-2 antigens were detected on mature adipocytes and a proportion of non-lipid-containing cells in stromal-vascular cultures. Adipocytes and associated capillary networks in subcutaneous adipose tissues as well as capillaries between the underlying muscle fiber bundles bound each antibody, whereas the AD-2 monoclonal antibody also reacted with vessels but not capillaries in liver tissues. In stromal-vascular cell cultures prepared from newborn pig subcutaneous tissue, the AD-1 and AD-2 antibodies exhibited reactivity towards 45 percent and 10 percent respectively, of cells 24 hours after seeding. On the other hand, only 4 percent and 1 percent of the cells in cultures prepared from 60 day fetal subcutaneous tissues expressed detectable amounts of the AD-1 and AD-2 antigens, respectively. In conclusion, cells along the adipogenic lineage possess cell surface antigens which may not be unique to adipogenic cells, but do exhibit differential expression among cell populations within adipose tissues. A temporal relationship between adipogenesis and angiogenesis was also demonstrated.

Adipose Tissue↗
PubMed1989

Fetal hypophysectomy causes a decrease in preadipocyte growth and insulin like growth factor-1 in pigs.

Fetal pigs in one uterine horn of each of five gilts were hypophysectomized (HX) in utero by electrical cauterization at 72-74 days of gestation and sera collected at 110 days of gestation. Sera from HX fetuses had lower levels of insulin-like growth factor-1 compared to control littermates (P less than .05). Sera were tested for their effects on primary cultures of stromal-vascular cells from adipose tissue. The soluble protein concentration/dish was lower when pig cells were cultured in sera from HX fetuses compared to sera from control fetuses (P less than .01). Sera from HX fetuses inadequately supported growth of stromal-vascular cells so subsequent experiments utilized pooled sera from normal and HX adult pigs. Sera from HX and control fetuses were mixed with sera from the two adult pools and tested for incorporation of tritiated thymidine into rat preadipocytes and the appearance of adipocytes (determined histochemically) in pig stromal-vascular cultures. In cultures fed sera from HX fetuses there was a lower (P less than .05) number of pig fat cells/culture and a lower level (P less than .06) of preadipocyte proliferation in rat cell cultures when compared to control fetal sera. Fetal pig serum contains factors (adipogenic) which promote the proliferation and differentiation of adipocytes in culture. Serum from HX fetuses has a lower level of adipogenic factors.

Adipose Tissue↗
PubMed1989

Primary cultures of stromal-vascular cells from pig adipose tissue: the influence of glucocorticoids and insulin as inducers of adipocyte differentiation.

Porcine stromal-vascular (S-V) cells from perirenal and subcutaneous fat depots were studied in a primary culture system. The ability of S-V cells to differentiate into adipocytes in response to insulin and to glucocorticoid treatments was determined. Treatments were either hydrocortisone plus insulin, dexamethasone plus isobutylmethylxanthine, or insulin. Insulin did not affect glycerol phosphate dehydrogenase (GPDH) activity or cytodifferentiation in porcine S-V cultures. All cultures differentiated in response to glucocorticoid addition. Glycerol phosphate dehydrogenase specific activity increased an average of 42.5% and the number of cell clusters with multilocular fat cells markedly increased. Cells receiving insulin after dexamethasone-stimulated differentiation had no greater ability to respond to the insulin than cells from control cultures. Glucocorticoids promoted cytodifferentiation of porcine S-V cells to adipocytes in a qualitative manner similar to other preadipocyte models; quantitatively, however, the response was greatly diminished.

1-Methyl-3-isobutylxanthine↗