Estimating risk of endometrial malignancy and other intracavitary uterine pathology in women without abnormal uterine bleeding using IETA-1 multinomial regression model: validation study.

OBJECTIVES: To assess the ability of the International Endometrial Tumor Analysis (IETA)-1 polynomial regression model to estimate the risk of endometrial cancer (EC) and other intracavitary uterine pathology in women without abnormal uterine bleeding. METHODS: This was a retrospective study, in which we validated the IETA-1 model on the IETA-3 study cohort (n = 1745). The IETA-3 study is a prospective observational multicenter study. It includes women without vaginal bleeding who underwent a standardized transvaginal ultrasound examination in one of seven ultrasound centers between January 2011 and December 2018. The ultrasonography was performed either as part of a routine gynecological examination, during follow-up of non-endometrial pathology, in the work-up before fertility treatment or before treatment for uterine prolapse or ovarian pathology. Ultrasonographic findings were described using IETA terminology and were compared with histology, or with results of clinical and ultrasound follow-up of at least 1 year if endometrial sampling was not performed. The IETA-1 model, which was created using data from patients with abnormal uterine bleeding, predicts four histological outcomes: (1) EC or endometrial intraepithelial neoplasia (EIN); (2) endometrial polyp or intracavitary myoma; (3) proliferative or secretory endometrium, endometritis, or endometrial hyperplasia without atypia; and (4) endometrial atrophy. The predictors in the model are age, body mass index and seven ultrasound variables (visibility of the endometrium, endometrial thickness, color score, cysts in the endometrium, non-uniform echogenicity of the endometrium, presence of a bright edge, presence of a single dominant vessel). We analyzed the discriminative ability of the model (area under the receiver-operating-characteristics curve (AUC); polytomous discrimination index (PDI)) and evaluated calibration of its risk estimates (observed/expected ratio). RESULTS: The median age of the women in the IETA-3 cohort was 51 (range, 20-85) years and 51% (887/1745) of the women were postmenopausal. Histology showed EC or EIN in 29 (2%) women, endometrial polyps or intracavitary myomas in 1094 (63%), proliferative or secretory endometrium, endometritis, or hyperplasia without atypia in 144 (8%) and endometrial atrophy in 265 (15%) women. The endometrial sample had insufficient material in five (0.3%) cases. In 208 (12%) women who did not undergo endometrial sampling but were followed up for at least 1 year without clinical or ultrasound signs of endometrial malignancy, the outcome was classified as benign. The IETA-1 model had an AUC of 0.81 (95% CI, 0.73-0.89, n = 1745) for discrimination between malignant (EC or EIN) and benign endometrium, and the observed/expected ratio for EC or EIN was 0.51 (95% CI, 0.32-0.82). The model was able to categorize the four histological outcomes with considerable accuracy: the PDI of the model was 0.68 (95% CI, 0.62-0.73) (n = 1532). The IETA-1 model discriminated very well between endometrial atrophy and all other intracavitary uterine conditions, with an AUC of 0.96 (95% CI, 0.95-0.98). Including only patients in whom the endometrium was measurable (n = 1689), the model's AUC was 0.83 (95% CI, 0.75-0.91), compared with 0.62 (95% CI, 0.52-0.73) when using endometrial thickness alone to predict malignancy (difference in AUC, 0.21; 95% CI, 0.08-0.32). In postmenopausal women with measurable endometrial thickness (n = 848), the IETA-1 model gave an AUC of 0.81 (95% CI, 0.71-0.91), while endometrial thickness alone gave an AUC of 0.70 (95% CI, 0.60-0.81) (difference in AUC, 0.11; 95% CI, 0.01-0.20). CONCLUSION: The IETA-1 model discriminates well between benign and malignant conditions in the uterine cavity in patients without abnormal bleeding, but it overestimates the risk of malignancy. It also discriminates well between the four histological outcome categories. © 2023 International Society of Ultrasound in Obstetrics and Gynecology.

Benign descriptors and ADNEX in two-step strategy to estimate risk of malignancy in ovarian tumors: retrospective validation in IOTA5 multicenter cohort.

OBJECTIVE: Previous work has suggested that the ultrasound-based benign simple descriptors (BDs) can reliably exclude malignancy in a large proportion of women presenting with an adnexal mass. This study aimed to validate a modified version of the BDs and to validate a two-step strategy to estimate the risk of malignancy, in which the modified BDs are followed by the Assessment of Different NEoplasias in the adneXa (ADNEX) model if modified BDs do not apply. METHODS: This was a retrospective analysis using data from the 2-year interim analysis of the International Ovarian Tumor Analysis (IOTA) Phase-5 study, in which consecutive patients with at least one adnexal mass were recruited irrespective of subsequent management (conservative or surgery). The main outcome was classification of tumors as benign or malignant, based on histology or on clinical and ultrasound information during 1 year of follow-up. Multiple imputation was used when outcome based on follow-up was uncertain according to predefined criteria. RESULTS: A total of 8519 patients were recruited at 36 centers between 2012 and 2015. We excluded patients who were already in follow-up at recruitment and all patients from 19 centers that did not fulfil our criteria for good-quality surgical and follow-up data, leaving 4905 patients across 17 centers for statistical analysis. Overall, 3441 (70%) tumors were benign, 978 (20%) malignant and 486 (10%) uncertain. The modified BDs were applicable in 1798/4905 (37%) tumors, of which 1786 (99.3%) were benign. The two-step strategy based on ADNEX without CA125 had an area under the receiver-operating-characteristics curve (AUC) of 0.94 (95% CI, 0.92-0.96). The risk of malignancy was slightly underestimated, but calibration varied between centers. A sensitivity analysis in which we expanded the definition of uncertain outcome resulted in 1419 (29%) tumors with uncertain outcome and an AUC of the two-step strategy without CA125 of 0.93 (95% CI, 0.91-0.95). CONCLUSION: A large proportion of adnexal masses can be classified as benign by the modified BDs. For the remaining masses, the ADNEX model can be used to estimate the risk of malignancy. This two-step strategy is convenient for clinical use. © 2022 The Authors. Ultrasound in Obstetrics & Gynecology published by John Wiley & Sons Ltd on behalf of International Society of Ultrasound in Obstetrics and Gynecology.

Diagnostic accuracy of transvaginal ultrasound for detection of endometriosis using International Deep Endometriosis Analysis (IDEA) approach: prospective international pilot study.

OBJECTIVE: To evaluate the diagnostic accuracy of transvaginal ultrasound (TVS) in predicting deep endometriosis (DE) following the International Deep Endometriosis Analysis (IDEA) consensus methodology. METHODS: This was an international multicenter prospective diagnostic accuracy study involving eight centers across six countries (August 2018-November 2019). Consecutive participants with endometriosis suspected based on clinical symptoms or historical diagnosis of endometriosis were included. The index test was TVS performed preoperatively in accordance with the IDEA consensus statement. At each center, the index test was interpreted by a single sonologist. Reference standards were: (1) direct visualization of endometriosis at laparoscopy, as determined by a non-blinded surgeon with expertise in endometriosis surgery; and (2) histological assessment of biopsied/excised tissue. Surgery was performed within 12 months following the index TVS. Accuracy, sensitivity, specificity, positive and negative predictive values (PPV and NPV) and positive and negative likelihood ratios (LR+ and LR-) of TVS in the diagnosis of DE were calculated. RESULTS: Included in the study were 273 participants with complete clinical, TVS, laparoscopic and histological data. Of these, based on histology, 256 (93.8%) were confirmed to have endometriosis, including superficial endometriosis, and 190 (69.6%) were confirmed to have DE. Based on surgical visualization, 207/273 (75.8%) patients had DE. For DE overall, the diagnostic performance of TVS based on surgical visualization as the reference standard was as follows: accuracy, 86.1%; sensitivity, 88.4%; specificity, 78.8%; PPV, 92.9%; NPV, 68.4%; LR+, 4.17; LR-, 0.15, and the diagnostic performance of TVS based on histology as the reference standard was as follows: accuracy, 85.9%; sensitivity, 89.8%; specificity, 75.9%; PPV, 90.4%; NPV, 74.6%; LR+, 3.72; LR-, 0.13. CONCLUSIONS: Using the IDEA consensus methodology provides strong diagnostic accuracy for TVS assessment of DE. We found a higher TVS detection rate of DE overall than that reported by the most recent meta-analysis on the topic (sensitivity, 79%), albeit with a lower specificity. © 2022 International Society of Ultrasound in Obstetrics and Gynecology.

Ultrasound features of endometrial pathology in women without abnormal uterine bleeding: results from the International Endometrial Tumor Analysis study (IETA3).

OBJECTIVES: The primary aim of this study was to describe the ultrasound features of various endometrial and other intracavitary pathologies in women without abnormal uterine bleeding (AUB) using the International Endometrial Tumor Analysis (IETA) terminology. The secondary aim was to compare our findings with published data on women with AUB. METHODS: This was a prospective observational study of women presenting at one of seven centers specialized in gynecological ultrasonography, from 2011 until 2018, for indications unrelated to AUB. All patients underwent transvaginal ultrasound using the IETA examination and measurement techniques. Ultrasonography was performed as part of routine gynecological examination or follow-up of non-endometrial pathology, or as part of the work-up before undergoing treatment for infertility, uterine prolapse or ovarian pathology. Ultrasound findings were described using the IETA terminology. Endometrial sampling was performed after the ultrasound scan. The histological endpoints were endometrial atrophy, proliferative or secretory endometrium, endometrial hyperplasia without atypia, endometrial polyp, intracavitary leiomyoma, endometrial intraepithelial neoplasia (EIN), endometrial cancer (EC) and insufficient tissue. The findings in our cohort of women without AUB were compared with those in a published cohort of women with AUB who were examined with transvaginal ultrasound between 2012 and 2015 using the same IETA examination technique and terminology. RESULTS: In this study (IETA3), we included 1745 women without AUB who underwent a standardized transvaginal ultrasound examination followed by either endometrial sampling with histological diagnosis (n = 1537) or at least 1 year of clinical and ultrasound follow-up (n = 208). Of these, 858 (49.2%) women were premenopausal and 887 (50.8%) were postmenopausal. Histology showed the presence of EC and/or EIN in 29 (1.7%) women, endometrial polyps in 1028 (58.9%), intracavitary myomas in 66 (3.8%), proliferative or secretory changes or hyperplasia without atypia in 144 (8.3%), endometrial atrophy in 265 (15.2%) and insufficient tissue in five (0.3%). Most cases of EC or EIN (25/29 (86.2%)) were diagnosed after menopause. The mean endometrial thickness in women with EC or EIN was 11.2 mm (95% CI, 8.9-13.6 mm), being on average 2.4 mm (95% CI, 0.3-4.6 mm) thicker than their benign counterparts. Women with malignant endometrial pathology manifested more frequently non-uniform echogenicity (22/29 (75.9%)) than did those with benign endometrial pathology (929/1716 (54.1%)) (difference, +21.8% (95% CI, +4.2% to +39.2%)). Moderate to abundant vascularization (color score 3-4) was seen in 31.0% (9/29) of cases with EC or EIN compared with 12.8% (220/1716) of those with a benign outcome (difference, +18.2% (95% CI, -0.5% to +36.9%)). Multiple multifocal vessels were recorded in 24.1% (7/29) women with EC or EIN vs 4.0% (68/1716) of those with a benign outcome (difference, +20.2% (95% CI, +4.6% to +35.7%)). A regular endometrial-myometrial junction was seen less frequently in women with EC or EIN (19/29 (65.5%)) vs those with a benign outcome (1412/1716 (82.3%)) (difference, -16.8% (95% CI, -34.2% to +0.6%)). In women with endometrial polyps without AUB, a single dominant vessel was the most frequent vascular pattern (666/1028 (64.8%)). In women with EC, both in those with and those without AUB, the endometrium usually manifested heterogeneous echogenicity, but the endometrium was on average 8.6 mm (95% CI, 5.2-12.0 mm) thinner and less intensely vascularized (color score 3-4: difference, -26.8% (95% CI, -52.2% to -1.3%)) in women without compared to those with AUB. In both pre- and postmenopausal women, asymptomatic endometrial polyps were associated with a thinner endometrium, and they manifested more frequently a bright edge, a regular endometrial-myometrial junction and a single dominant vessel than did polyps in symptomatic women, and they were less intensely vascularized. CONCLUSIONS: We describe the typical ultrasound features of EC, polyps and other intracavitary histologies using IETA terminology in women without AUB. Our findings suggest that the presence of asymptomatic polyps or endometrial malignancy may be accompanied by thinner and less intensely vascularized endometria than their symptomatic counterparts. © 2022 International Society of Ultrasound in Obstetrics and Gynecology.

Learning curve for ultrasonographic diagnosis of deep infiltrating endometriosis using structured offline training program.

OBJECTIVE: To assess the learning curves of trainees during a structured offline/hands-on training program for the ultrasonographic diagnosis of deep infiltrating endometriosis (DIE). METHODS: Four trainees (all Ob/Gyn postgraduates with at least 5 years' experience in ultrasonography in obstetrics and gynecology, but with no experience of sonographic examination of DIE) participated in the study. They underwent a 2-week training program with a single trainer. Day 1 was devoted to theoretical issues and guided offline analysis of 10 three-dimensional ultrasound volumes. During the following days, four sessions of real-time sonographic examinations were performed in a DIE referral center ultrasound unit. In between these sessions, the trainees analyzed four datasets offline, each containing 25 volumes. At the end of each set, misinterpreted volumes were reassessed with the trainer. Presence or absence of DIE at surgery was considered the gold standard. The trainees' learning process was evaluated by learning-curve cumulative summation (LC-CUSUM) and the deviations of the trainees' level of performance at the control stage was assessed by CUSUM (standard CUSUM), for different locations of DIE. RESULTS: The trainees reached competence after an average of 17 (range, 14-21) evaluations for bladder, 40 (range, 30-60) for rectosigmoid, 25 (range, 14-34) for forniceal, 44 (range, 25-66) for uterosacral ligament (USL) and 21 (range, 14-43) for rectovaginal septum (RVS) locations of DIE, and then kept the process under control, with error levels of less than 4.5% until the end of the test. The overall accuracy for each trainee in diagnosis of DIE at the different locations ranged from 0.91 to 0.98 for bladder DIE, from 0.80 to 0.94 for rectosigmoid DIE, from 0.90 to 0.94 for forniceal DIE, from 0.79 to 0.82 for USL DIE and from 0.89 to 0.98 for RVS DIE. CONCLUSIONS: The suggested 2-week training program, based on a mixture of offline and live scanning sessions, is feasible and apparently provides effective training for the ultrasonographic diagnosis of DIE. Copyright © 2018 ISUOG. Published by John Wiley & Sons Ltd.

Long-term feed intake regulation in sheep is mediated by opioid receptors.

These experiments were conducted to determine if 1) syndyphalin-33 (SD33), a mu-opioid receptor ligand, affects feed intake; 2) SD33 effects on feed intake are mediated by actions on opioid receptors; and 3) its activity can counteract the reduction in feed intake associated with administration of bacterial endotoxin. In Exp. 1, 5 mixed-breed, castrate male sheep were housed indoors in individual pens. Animals had ad libitum access to water and concentrate feed. Saline (SAL; 0.9% NaCl) or SD33 (0.05 or 0.1 micromol/kg of BW) was injected i.v., and feed intake was determined at 2, 4, 6, 8, 24, and 48 h after the i.v. injections. Both doses of SD33 increased (at least P < 0.01) feed intake at 48 h relative to saline. In Exp. 2, SAL + SAL, SAL + SD33 (0.1 micromol/kg of BW), naloxone (NAL; 1 mg/kg of BW) + SAL, and NAL + SD33 were injected i.v. Food intake was determined as in Exp. 1. The SAL + SD33 treatment increased (P = 0.022) feed intake at 48 h relative to SAL + SAL. The NAL + SAL treatment reduced (at least P < 0.01) feed intake at 4, 6, 8, 24, and 48 h, whereas the combination of NAL and SD33 did not reduce feed intake at 24 (P = 0.969) or 48 h (P = 0.076) relative to the saline-treated sheep. In Exp. 3, sheep received 1 of 4 treatments: SAL + SAL, SAL + 0.1 micromol of SD33/kg of BW, 0.1 microg of lipopolysaccharide (LPS)/kg of BW + SAL, or LPS + SD33, and feed intake was monitored as in Exp. 1. Lipopolysaccharide suppressed cumulative feed intake for 48 h (P < 0.01) relative to saline control, but SD33 failed to reverse the reduction in feed intake during this period. These data indicate that SD33 increases feed intake in sheep after i.v. injection, and its effects are mediated via opioid receptors. However, the LPS-induced suppression in feed intake cannot be overcome by the opioid receptor ligand, SD33.

Lysine requirement of finishing pigs administered porcine somatotropin by sustained-release implant.

To alleviate the need for daily injection of porcine somatotropin (pST), a sustained-release implant (pSTSR) was devised that continuously delivers a daily dose of 2 mg of pST for 42 d. Ninety-six white composite (Large White x Landrace) finishing barrows (83.6 +/- 1.2 kg BW) were assigned to receive zero or two pSTSR implants (4 mg pST/d) and to consume one of six diets differing in total Lys concentration (0.29, 0.52, 0.75, 0.98, 1.21, or 1.44%, as-fed basis). Diets were formulated to be isocaloric and based on the ideal protein concept. Pigs were housed individually, allowed ad libitum access to feed and water, and slaughtered at 112 kg of BW. The pSTSR affected neither ADG (P = 0.88) nor 10th rib LM area (LMA; P = 0.51), but it decreased (P < 0.01) ADFI, average backfat thickness, 10th rib fat depth, weights of leaf fat and ham fat, improved (P < 0.05) G:F, and increased (P < 0.01) weights of four trimmed lean cuts (T-cuts), and percentages of ham lean and bone. Increasing total Lys increased ADG (quadratic; P < 0.05) and ADFI (linear; P < 0.01). The G:F, plasma urea N concentrations (PUN), and T-cuts were affected by the interaction pSTSR x dietary Lys (P < 0.01). Without pSTSR, the G:F did not differ (P = 0.37) among pigs fed 0.52% and greater total Lys. With pSTSR, the G:F was less (P < 0.05) for pigs fed 0.52% than 0.98 and 1.44% total Lys. Increases in dietary total Lys resulted in increased PUN (P < 0.01), and incremental increases were less in pSTSR-implanted pigs. Maximal yield of T-cuts was at 0.98% dietary total Lys in nonimplanted pigs and 1.21% total Lys in pSTSR-implanted pigs. Estimates of total Lys requirements of pigs without and with pSTSR, respectively, were 0.52 and 0.86% for growth (ADG and G:F) and 0.73 and 0.88% for lean production (LMA and T-cuts). Equivalent apparent ileal digestible Lys requirements of pigs without and with pSTSR, respectively, were 0.44 and 0.68% for growth, and 0.62 and 0.75% for lean production. With ADFI of 3.5 kg daily, an intake of approximately 26.1 g of total daily Lys (0.75%) or 22.4 g of apparent ileal digestible Lys is needed to maximize lean production in finishing barrows receiving 4 mg pST/d via sustained-release implant.

Intracerebroventricular melanin-concentrating hormone stimulates food intake in sheep.

Melanin-concentrating hormone (MCH) stimulates feeding when injected intracerebroventricularly (ICV) in rats. At present it is not clear whether the function of MCH is similar in ruminants, which are species with a continuous delivery of nutrients. Therefore the current investigation sought to determine the role of MCH in sheep. In the first experiment, six, castrate male sheep were satiated and received one of four treatments [saline, 0.1, or 1.0 nmol/kg MCH, and NPY (0.1 nmol/kg)] injected ICV over 30s, then infused ICV for 6 h ( approximately 500 microl/h). Food intake was measured for 2 h before and at 2, 4, 6, 8, 12 and 24 h. In this experiment, feed intake was increased (P

Effect of intracerebroventricular orexin-B on food intake in sheep.

Orexin is a hypothalamic neuropeptide that regulates feeding behavior in rats. Orexin-B has recently been cloned in pigs and was shown to stimulate food intake after intramuscular injection. This study was designed to determine whether intracerebroventricular (ICV) and intravenous injections of orexin could regulate appetite in sheep. Suffolk wethers were moved to indoor facilities, adapted to diets for 6 wk, and trained to stand in stanchions for 3 to 6 h each day for 2 wk before indwelling ICV cannulas were installed. These sheep were provided water and they consumed feed ad libitum. On the day before an experiment, each sheep was cannulated in a jugular vein. On the day of an experiment, sheep were placed in stanchions and allowed to stand for 1 h before use. Sheep were then monitored over a 2-h control period before i.v. injection with saline or porcine orexin-B (3 micrograms/kg BW) or ICV injection with artificial cerebrospinal fluid (CSF), orexin (0.03, 0.3, or 3 micrograms/kg BW) or in a second experiment with either orexin B (0.03, 0.3, 3 micrograms/kg BW), neuropeptide-Y (NPY; 0.3 microgram/kg BW), or orexin plus NPY. Food intake was monitored for consecutive 2-h periods. The i.v. injections of orexin did not affect food intake or metabolite or hormone concentrations. In ICV sheep, orexin increased food intake at 2 (P < 0.04) and at 4 h (P < 0.02). Food intake was greatest with the 0.3 microgram/kg BW dosage of orexin (P < 0.05). In the first 2 h after injection, orexin had an effect similar to that of NPY (0.23 kg for orexin and 0.2 kg for NPY). The combination of NPY and orexin had a greater effect on food intake (to 0.34 kg) than did either orexin (P < 0.05) or NPY (P < 0.008) alone. Differences were not apparent in the subsequent 2-h interval. No differences were noted in free fatty acid, glucose, growth hormone, luteinizing hormone, or insulin concentrations following orexin injection. There was an effect of ICV orexin treatment on plasma cortisol concentrations (P < 0.002). Cortisol was increased by orexin at the 0- to 2-h (P < 0.008) and in the 2- to 4-h (P < 0.009) intervals after orexin injection. These data indicate that central administration of orexin stimulates feed intake in sheep.

The influence of recombinant bovine somatotropin on dietary energy level-related growth of Holstein-Friesian bull calves.

Our objective for this study was to assess the effect of recombinant bovine somatotropin (rbST) in overcoming the biological effects attributed to live weight increase and age on growth and fat deposition in male cattle. Holstein-Friesian bull calves (n = 56; 182.2 +/- 14.7 d old) were allotted to four subtreatments in a randomized complete block with a factorial arrangement of two levels of rbST: 0 and 500 mg of Posilac, every 2 wk, and two dietary metabolizable energy (ME) concentrations: low metabolizable energy (LME) and high metabolizable energy (HME); 10 vs 11.3 MJ/kg DM, respectively. The effect of rbST treatment on daily gain was expressed mostly on the HME diets. The rbST treatment had no effect on the animals fed the LME diets before the age of 240 d was reached. Dry matter intake and the effect of rbST treatment on DM intake were inversely related to the energy concentration of the diet. The degree of fatness of the animals was significantly reduced by rbST treatment and significantly increased by energy concentration of the diet. The major effect of rbST, under the experimental conditions, in regards to adipose tissue deposition, was on the fat depots and not on the intramuscular fat. The concentration of bST, IGF-I, and insulin in the plasma was increased (P < .001) owing to rbST treatment. Lower metabolizable energy supply led to a higher (P < .001) plasma bST concentration, nonsignificantly lower plasma IGF-I and thyroid hormone concentrations, and lower (P < .001) plasma insulin concentration. A trend (P = .065) toward an increase in PUFA was found in the muscle of the rbST-treated and the HME diet animals. At a young age, when the natural growth potential is high, rbST treatment will be efficient only when a diet allowing a high digestible energy intake is provided.

Alterations in the growth hormone/insulin-like growth factor I pathways in feline GM1 gangliosidosis.

Cats affected with feline GM1 gangliosidosis, an autosomal, recessively inherited, lysosomal enzymopathy, have progressive neurological dysfunction, premature thymic involution, stunted growth, and premature death. Although increased membrane GM1 gangliosides can result in increased apoptosis of thymocytes, there is not a direct correlation between thymocyte surface GM1 and thymic apoptosis in vivo, suggesting that other factors may be important to the pathogenesis of thymic involution in affected cats. Because GH and insulin-like growth factor I (IGF-I) are important hormonal peptides supporting thymic function and affecting growth throughout the body, particularly in the prepubescent period, several components of the GH/IGF-I pathway were compared in GM1 mutant and normal age-matched cats. GM1 mutant cat serum IGF-I concentrations were reduced significantly compared with those in normal cats by 150 days of age, and GM1 mutant cats had no peripubertal increase in serum IGF-I. Additionally, IGF-binding protein-3 was reduced, and IGF-binding protein-2 was elevated significantly in GM1 mutant cats more than 200 days of age. Liver IGF-I messenger RNA and pituitary GH messenger RNA both were reduced significantly in GM1 mutant cats. After stimulation by exogenous recombinant canine GH, serum IGF-I levels increased significantly in GM1 mutant cats, indicating that GH/IGF-I signaling pathways within the liver remain intact and suggesting that alterations are external to the liver.

Interactions between environmental temperature and porcine growth hormone (pGH) treatment in neonatal pigs.

Age-dependent interactions between environmental temperature and porcine growth hormone (pGH) treatment on the function of the somatotrophic axis were evaluated in the neonatal pig. At 3 d of age, 40 Landrace x Yorkshire x Duroc piglets received intraperitoneal implants containing either recombinant pGH (0.5 mg/d; n = 20) or vehicle (control; n = 20). Piglets were maintained at either a low (21 degrees C, 50% relative humidity; n = 20) or high (32 degrees C, 50% relative humidity; n = 20) temperature. At 4 and 6 wk of age, 5 pGH-treated and 5 control piglets from each thermal group were sacrificed for tissue collection. Blood samples were collected at the time of sacrifice and analyzed for serum concentrations of GH, insulin-like growth factor 1 (IGF-1), and IGF-2. Liver RNA was analyzed for mRNAs specific for the GH receptor, IGF-1, IGF-2, and IGF binding protein 3. There was no effect of pGH treatment (P = 0.4) on average daily gain; however, both age (P = 0.002) and temperature (P = 0.001) had an effect on average daily gain such that those animals maintained at a low temperature and those sacrificed at 6 wk had greater average daily gains. Serum concentration of GH was elevated (P = 0.003) by pGH treatment and was lowest in the 6-wk-old group (P = 0.008). Serum concentration of IGF-1 was elevated (P = 0.007) by pGH treatment and increased with age (P = 0.01). Liver GH receptor mRNA was unaffected (P > 0.5) by pGH treatment, but was greater in the 6-wk-old group (P < 0.0001) and in piglets maintained at the high temperature (P = 0.04). IGF-1 mRNA was enhanced by pGH treatment (P = 0.0003) and by exposure to the high temperature (P = 0.04), but did not differ (P > 0.5) between age groups. IGF-2 mRNA was greater (P = 0.0009) in the 4-wk-old group and in piglets maintained at the high temperature (P = 0.007), but was unaffected (P = 0.5) by pGH treatment. IGF binding protein 3 mRNA increased with age (P = 0.0004) and was stimulated by pGH treatment in the 6-wk-old group (P = 0.034). The relatively lower level of GH receptor and IGF mRNAs in conjunction with greater growth in the cold environment suggests that somatotrophic gene expression in the liver is not rate limiting for growth in the neonatal pig.

Growth, body composition, and endocrine responses to chronic administration of insulin-like growth factor I and(or) porcine growth hormone in pigs.

The actions of IGF-I, alone and in combination with porcine growth hormone (pGH), on growth and circulating endocrines and metabolites important in growth were investigated in peripubertal-age Meishan barrows. Pigs were assigned to four treatments: control, buffer; IGF-I, 33 microg rhIGF-I/kg BW injected twice daily; pGH, 33 microg rpGH/kg BW injected once daily; and IGF-I+pGH, 33 microg rhIGF-I/kg BW injected twice daily plus 33 microg rpGH/kg BW injected once daily. Treatments were administered for 28 d. Feed intake, BW, and backfat were recorded and blood samples were collected weekly. At slaughter, organ and primal cut weights were recorded. Offal and half the carcass were ground for chemical analysis. Serum concentrations of IGF-I on d 7, 14, 21, and 28 in the IGF-I, pGH, and IGF-I+pGH groups were increased 60, 107, and 131%, respectively, compared with those of the control group. Administration of pGH increased gain 43%, feed efficiency 60%, carcass protein accretion 88%, and trimmed lean cuts 16%, whereas IGF-I administration increased gain 22%, carcass protein accretion 33%, and trimmed lean cuts 5%. There was little difference in responses to administration of IGF-I+pGH and pGH alone except that coadministration of IGF-I with pGH reduced the ability of pGH to suppress backfat gain (P < .02). Even though administration of IGF-I resulted in a 60% increase in chronic nadir serum concentrations of IGF-I, only a few growth and carcass measures were changed when compared with control pigs. These included increased (P < .05) weight of body, leaf fat, kidneys, and belly. The actions of pGH on growth of pigs were not mimicked, and some were countermanded by administration of IGF-I at a dose that produces significantly increased serum concentrations of IGF-I.

Radiographic, densitometric, and biomechanical effects of recombinant canine somatotropin in an unstable ostectomy gap model of bone healing in dogs.

OBJECTIVE: To determine the effect of recombinant canine somatotropin (STH) on radiographic, densitometric, and biomechanical aspects of bone healing using an unstable ostectomy gap model. STUDY DESIGN: After an ostectomy of the midshaft radius, bone healing was evaluated over an 8-week period in control dogs (n = 4) and dogs receiving recombinant canine STH (n = 4). ANIMALS OR SAMPLE POPULATION: Eight sexually intact female Beagle dogs, 4 to 5 years old. METHODS: Bone healing was evaluated by qualitative and quantitative evaluation of serial radiographs every 2 weeks. Terminal dual-energy x-ray absorptiometry and three-point bending biomechanical testing were also performed. RESULTS: Dogs receiving STH had more advanced radiographic healing of ostectomy sites. Bone area, bone mineral content, and bone density were two to five times greater at the ostectomy sites of treated dogs. Ultimate load at failure and stiffness were three and five times greater in dogs receiving STH. CONCLUSIONS: Using the ostectomy gap model, recombinant canine STH enhanced the radiographic, densitometric, and biomechanical aspects of bone healing in dogs. CLINICAL RELEVANCE: Dogs at risk for delayed healing of fractures may benefit from treatment with recombinant canine STH.

The effects of dietary restriction on insulin-like growth factor (IGF)-I and II, and IGF-binding proteins in chickens.

The effect of feed (energy/protein) restriction on circulating concentrations of insulin-like growth factor (IGF)-I and II, and IGF-binding proteins (IGFBPs) was examined in young (4-week-old) chickens. Increasing levels of feed restriction caused progressive growth retardation, as evidenced by decreased body-weight gain and reduced bone growth. Plasma concentrations of both IGF-I and IGF-II were decreased, and the degree of reduction in the plasma concentrations of these growth factors appeared to be related to the magnitude of feed restriction. A tendency for greater decreases in these growth factors appeared to be associated with greater feed restriction at the majority of time points evaluated. However, nutritional restriction had a greater effect on plasma concentrations of IGF-I than on those of IGF-II. The reductions in plasma concentrations of IGF-I were observed earlier in the experiment and at a lower degree of nutritional deprivation than for plasma concentrations of IGF-II, possibly suggesting greater sensitivity of IGF-I plasma concentrations to feed restriction. Three IGFBPs with molecular weights of 30, 36, and 40 kDa were detected by radioligand assay following separation by SDS-electrophoresis. The 30-kDa IGFBP was most affected by feed restriction with binding activity of this IGFBP increased by 2 days of feed restriction irrespective of the degree of feed deprivation. The binding activity of the 36-kDa IGFBP was increased, albeit transiently, on the second day of feed restriction. Nutritional restriction had no discernible effect on the binding activity of the 40-kDa IGFBP. Increases in the binding activity of the 30-kDa IGFBP appeared to correspond with the observed decreases in IGF-I plasma concentrations. This suggests decreased bioavailability of IGF-I, and possibly IGF-II, attributed to the formation of a complex between IGF-I and the 30-kDa IGFBP during feed restriction. The initial increase in binding activity of the 36-kDa IGFBP may suggest that this binding protein also plays a role in the regulation and availability of circulating concentrations of IGF-I and IGF-II. Although the binding activity of the 40-kDa IGFBP was unaffected by feed restriction, we can not exclude its importance in the regulation of IGF-I. The substantial binding activity of the 40-kDa IGFBP observed in this experiment suggests that it is one of the major chicken IGFBPs, and that its role in IGF-I regulation warrants further study.

Ontogeny of insulin-like growth factors (IGF-I and IGF-II) and IGF-binding proteins in the chicken following hatching.

The present study examined plasma concentrations of insulin-like growth factor (IGF)-I, IGF-II and IGF-binding proteins (IGFBPs) during posthatch growth and development in chickens. Three distinct proteins which bound 125I-IGF-I were observed irrespective of age or sex, these having apparent molecular weights of 22, 28, and 36 kDa. The major IGFBP present during much of the growth and development period was the 28-kDa form followed by the 36-kDa form. Plasma concentrations of IGF-II and of the 22-kDa IGFBP showed little ontogenic variation with the exception of elevated levels of the 22 kDa IGFBP in 1-day-old chicks. The circulating concentrations of IGF-I and of the 28-kDa IGFBP increased progressively between the time of hatching to reach a maximum at 6 weeks of age and subsequently declined to lower levels in adults. Somewhat similarly, the 36-kDa IGFBP increased during early pre- and posthatching growth to a maximum at 6 weeks of age. There were marked sex differences in circulating concentrations of IGF-I in young (4 week) and adult chickens and in the 36-kDa IGFBP in the adult, both being lower in females. Estrogen treatment of adult male chickens decreased the circulating concentrations of IGF-I together with the level of both the 28- and 36-kDa IGFBPs. Testosterone treatment had no effect on the circulating concentrations of either IGF-I or IGFBPs in adult female chickens. We conclude that the relative levels of IGF-I, IGF-II, and the IGFBPs change with age. In addition, circulating concentrations of estrogen may play a role in the regulation of IGF-I and IGFBPs.

Ontogenic changes in the circulating concentrations of insulin-like growth factor (IGF)-I, IGF-II, and IGF-binding proteins in the chicken embryo.

The ontogeny of circulating concentrations of insulin-like growth factor (IGF-)-I, IGF-II, and IGF-binding proteins (IGF-BPs) was examined in the chick embryo. Distinct ontogenic changes in the circulating concentrations of IGF-I and IGF-II were observed. The present study confirms the overall profile for circulating concentrations of IGF-I. During middevelopment, plasma concentrations of IGF-I increased to a maximum which was attained on Day 14.5 of incubation. Thereafter, plasma concentrations of IGF-I declined with decreases (P < 0.05) between Days 14.5 and 15.5 and between Days 16.5 and 17.5 of incubation. In contrast to the monophasic profile for IGF-I, plasma concentrations of IGF-II were maximal on Day 10.5 of incubation and declined to a nadir on Day 17.5 of incubation. In late developmental stages (17.5 or 18.5 days of incubation), three IGF-BPs, having molecular weights of 22, 28, and 36 kDa, were detected in the plasma of chick embryos. No significant ontogenic changes in the circulating levels of the 28- and 36-kDa IGF-BPs were observed. However, it should be noted that prior to Day 17.5 of incubation, the 22-kDa IGF-BP was nondetectable in the circulation. The role of these changes in the functioning of IGF in embryonic development is discussed.

Growth performance, endocrine, and metabolite responses of finishing hogs to porcine prolactin.

Prolactin, a member of the somatotropin-prolactin-placental lactogen gene family, increases feed intake and rate of weight gain in several species. To determine whether prolactin affects growth performance and carcass composition in swine, recombinant porcine prolactin (rpPRL) was administered to finishing hogs. Doses of 0, 2, 4, 8, and 16 mg of rpPRL/d and 4 mg of recombinant porcine somatotropin (rpST)/d were administered to groups of seven barrows and seven gilts initially weighing 75.0 +/- .2 kg for a 28-d period. Recombinant pPRL did not alter feed intake or growth rate or affect carcass composition. In addition, most growth-related blood variables did not change, although plasma IGF-I was increased in the 8 and 16 mg of rpPRL treatment groups. At slaughter, mammary development was apparent in rpPRL-treated gilts and was characterized by distended alveolar and ductal lumina and presence of secretory material. In rPST-treated hogs, feed intake was decreased 28% (P < .01), gain/feed was increased more in barrows than in gilts (59 vs 39%, treatment x sex interaction, P = .035), and growth rate was increased 22%, but in barrows only (treatment x sex interaction P = .005). Compared with those in control hogs, circulating concentrations of IGF-I, insulin, and glucose were 175, 311, and 22% higher, respectively, and of blood urea nitrogen were 62% lower in rpST-treated hogs (P < .05). These results suggest that rpPRL, at the doses administered, does not increase feed intake in finishing hogs in contrast to rats and other species.

Suppression of somatotroph function induced by growth hormone treatment in neonatal pigs.

The effect of recombinant porcine growth hormone (pGH) treatment on pituitary function was evaluated in young pigs. Piglets received intraperitoneal recombinant pGH implants (0.5 mg/d sustained release) or vehicle implants beginning at 3 d of age. Ten piglets were sacrificed at 4 and 6 wk of age (five piglets/treatment group) for the collection of pituitary glands, blood, and liver tissue. Blood samples also were drawn at 3 and 12 d of age. Serum concentrations of GH, prolactin (PRL), thyroid-stimulating hormone (TSH), insulin-like growth factor-1 (IGF-1) and IGF-2 were evaluated. Levels of IGF-1 and IGF-2 mRNA were determined in liver samples. Treatment with GH increased circulating levels of GH and IGF-1 (P < 0.01), but not PRL, TSH, or IGF-2. Hepatic IGF-1, but not IGF-2, mRNA levels were increased by pGH (P < 0.001). Cultured pituitary cells from each animal were challenged with 0.1, 1, and 10 nM GH-releasing hormone (GHRH); 2 nM 8-Br-cAMP; or 100 nM phorbol myristate acetate. The release of GH from cultured pituitary cells was stimulated by all secretagogues (P < 0.001). The secretion of GH, but not PRL or TSH, in culture was inhibited by previous in vivo GH treatment (P < 0.001). Similarly, cellular GH, but not PRL or TSH, content was lower in the GH-implant group (P = 0.005). Cell cultures from 6-wk-old piglets secreted more GH, but not PRL or TSH, than cultures from 4-wk-old piglets (P < 0.05). Likewise, cellular GH, but not PRL or TSH, content was greatest in cultures from 6-wk-old animals (P = 0.002). Piglet growth was not affected by exogenous GH treatment (P = 0.67). These results demonstrate that exogenous pGH treatment selectively down-regulates somatotroph function in young pigs.

Thymic, gonadal, and endocrine relationships in gilts and boars administered porcine somatotropin.

We hypothesized that much of the positive effect of porcine somatotropin (pST) on the immune system would be offset by the pST/IGF stimulation of gonadal function and that there would be negative effects on thymic weight and function from increased androgens in males. Male and female hogs (78 boars, 90 gilts) representing three genetic lines (lean, obese, and crossbred meat type) received 0, 2, or 4 mg of recombinant pST/d via implant for 42 d. Blood samples were collected on 0, 7, 14, 28, and 42 d of the trial for changes in pST, IGF-I, IGF-II, thymosin beta 4, dehydroepiandrosterone/dehydroepiandrosterone sulfate (DHEA/DHEASO4), and testosterone concentrations. Thymic, splenic, gonadal, and adrenal weights were collected at slaughter (d 42). Thymic weights increased with dose of pST in both sexes (P < .01), but splenic weights were unaltered. Adrenal weights increased with dose of pST (P < .01), but gonadal weights showed no response to pST in either sex or any line. Overall, concentrations of pST, IGF, and thymosin beta 4 increased with dose of pST. Serum testosterone concentrations in boars declined with dose of pST in lean and obese lines, and DHEA/ DHEASO4 declined in all lines with pST treatment. Testicular testosterone concentrations were not different among lines or doses of pST, indicating no stimulatory effect of increased pST/IGF-I on Leydig cell function. Although pST and its effects through increased circulating IGF are thought to be overall stimulants of growth and protein accretion, the actions on the tissues of the immune system (thymus/spleen) and steroidogenic tissues (adrenal/gonadal) can be selective. Increases in thymic weights and thymosin beta 4 concentrations from pST treatment in boars were partially due to the pST-induced decline in circulatory androgens, and the responses found in this in vivo model do not support pST/IGF effects documented in in vitro systems on Leydig cell function.