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The effect of the timing of posthatch feed restriction on adipose deposition and adipogenic gene expression in the broiler pectoralis major muscle was studied by applying a 20% feed restriction either the first or second week after hatch. Broiler chicks at hatch were divided into a full-fed (control) group and a 20% feed restriction group. The expression of adipogenic genes, peroxisome proliferator-activated receptor gamma (PPARγ), and CCAAT/enhancer-binding protein alpha (C/EBPα) were measured. The expression of both PPAR-γ and C/EBPα was affected by the wk 1 feed restriction with expression significantly increased during the first week posthatch. The deposition of fat within the pectoralis major muscle was affected by the timing of the feed restriction. Extensive fat depots were present by 27 d of age in the pectoralis major muscle of the wk 1 restricted group compared with the control. Fat deposition was eliminated when the 20% feed restriction occurred in wk 2. Taken together, these results demonstrate that the timing of early posthatch feed restrictions in chicks is critical in the deposition of fat in the pectoralis major muscle and expression of adipogenic genes. [ABSTRACT FROM AUTHOR]

The effect of the timing of an immediate posthatch feed restriction on broiler pectoralis major muscle development was studied by applying a 20% feed restriction either the first or second week after hatch. Pectoralis major muscle morphological structure and the expression of the myogenic transcriptional regulatory factors, myogenic determination factor 1 (MyoD), myogenic regulatory factor 4 (MRF4), and myogenin, were measured. Broiler chicks at hatch were divided into a fullfed (control) group and a 20% feed restriction treatment administered either the first or second week posthatch. At the end of the feed restriction, the chicks were placed on a full feed ad libitum diet with no further restrictions. Muscle fiber diameter and fiber bundle size of the pectoralis major muscle were smaller in the wk 1 restricted group than the control group by 7 d of age. By 15 d of age through the duration of the study, d 43, both endomysial and perimysial connective tissue spacing were diminished in the wk 1 feed restricted group. The expression of MyoD, MRF4, and myogenin was affected by the wk 1 feed restriction. The expression of MyoD and MRF4 was significantly increased during the first week posthatch. Both of the genes have been shown to be expressed during proliferation especially MyoD, which is required for muscle cell proliferation. In contrast, myogenin expression was significantly decreased. Myogenin expression is required for differentiation to occur. The morphological changes and gene expression changes observed with the wk 1 feed restriction were eliminated by moving the 20% feed restriction to wk 2, which is after the period of maximal myogenic satellite cell mitotic activity. Taken together, these results demonstrate that the timing of early posthatch feed restrictions to chicks is critical for the morphological development of the pectoralis major muscle and the expression of genes required for muscle satellite cell proliferation and differentiation. [ABSTRACT FROM AUTHOR]

It was apparent in previous studies at our institution using turkeys that measurements of muscle fibers and extracellular spacing were not adequate to explain what was observed in entire pectoralis major muscle sections. A rating system was developed in which muscle sections were rated from 1 (little extracellular matrix and indistinct muscle fibers) to 5 (large extracellular space and distinct muscle fibers). Maternal inheritance was observed at 16 wk of age but not at 8 or 20 wk of age. The purpose of the current study was to determine the effect of age on maternal inheritance. A line (F) selected long-term for increased 16-wk BW, its randombred control (RBC2), and reciprocal crosses between them were compared from 8 through 18 wk of age. Samples of pectoralis major muscle were obtained in a manner to avoid muscle contraction. After being fixed and cross-sectioned, the muscle samples were stained with hematoxylin and eosin and rated by 4 individuals. No significant difference among genetic groups was observed in scores at 8 wk of age. At 10 wk of age, the F line had lower scores than the other genetic groups. Maternal inheritance was suggested at 12 wk of age. The scores for RBC2 were higher than those for F, whereas the F x RBC2 cross did not differ from the pure RBC2 line score at this age. Although the RBC2 x F scores were higher than the pure F-line scores at 12 wk, they were lower than those of the F x RBC2 crosses. From 14 through 18 wk of age, the scores for the RBC2 line were higher than those for the F line and the maternal inheritance was absolute because the value for the individual crosses did not differ from that of the maternal parent. Based on the results, the type of mating used to produce commercial turkeys would have a major effect on breast muscle morphology from 12 through 18 wk of age. [ABSTRACT FROM AUTHOR]

Pectoralis major muscle morphology was studied in both sexes of a turkey line (E) selected long-term for increased egg production and its randombred control (RBC1) from 25 d of incubation through 20 wk posthatch. Pectoralis major muscle samples from 10 individuals from each line-sex-age subgroup were obtained in a manner to prevent contraction. The muscle samples were dehydrated, cleared, embedded in paraffin, sectioned, incubated, and rehydrated before staining with hematoxylin and eosin. Representative sections were given a score by 4 individuals based on breast muscle morphology. The scores ranged from 1 (little extracellular matrix and indistinct muscle fibers) to 5 (large extracellular space and distinct muscle fibers). Scores from 2 to 4 were intermediate to these extremes. The pectoralis major muscle morphology scores were highest at 25 d of incubation and declined greatly at 1 wk of age. The scores increased from 1 to 4 wk of age and remained constant through 20 wk of age. Males had higher scores than females. In the current study, there was no significant difference between the E and RBC1 lines. Based on the results of 3 experiments (2 published and the present one) using the E and RBC1 lines, it appears that genetic increases in egg production may be associated with a slight reduction in pectoralis major muscle morphology scores at 16 wk of age. [ABSTRACT FROM AUTHOR]

The membrane-associated heparan sulfate proteoglycan families consisting of the syndecans and glypicans are low-affinity receptors for fibroblast growth factor 2 (FGF2). Fibroblast growth factor 2 is a potent stimulator of skeletal muscle cell proliferation and a strong inhibitor of differentiation. Because syndecan-1, syndecan-4, and glypican-1 potentially play unique, but pivotal, roles in muscle cell proliferation and differentiation, these proteoglycans were examined for their effect on muscle cell proliferation and differentiation and FGF2 responsiveness. In the present study, turkey Randombred Control 2 line myogenic satellite cells were transfected with expression vector constructions of syndecan-1, syndecan-4, or glypican-1 to assay their role during muscle development and the effect on FGF2 responsiveness. During proliferation, only syndecan-1 increased proliferation. Both syndecan-4 and glypican-1 decreased proliferation at 72 h but generally did not affect the proliferation process. There was no interaction between the transfected gene and cell proliferation response to FGF2. Glypican-1 increased differentiation early in the process (24 h), and at later times differentiation was decreased by glypican-1. Both syndecan-1 and syndecan-4 overexpression decreased differentiation. During differentiation, except for glypican-1 at 48 h of differentiation, there was no interaction between gene treatment and FGF2 responsiveness. This result indicates that FGF2 responsiveness was not affected by the overexpression of syndecan-1, syndecan- 4, and glypican-1 during differentiation. These data demonstrate that syndecan-1, syndecan-4, or glypican-1 differentially affect the processes of turkey muscle cell proliferation and differentiation, and can regulate these developmental stages in an FGF2-independent manner. [ABSTRACT FROM AUTHOR]

Posthatch muscle growth is determined by the activation, differentiation, and fusion of satellite cells. Satellite cells composing an individual muscle are heterogeneous, which will differentially affect muscle growth. The proliferation and differentiation of turkey primary pectoralis major muscle cells were investigated in vitro at :1 d of age and at 4, 8, 16, 26, 35, 45, and 54 wk of age. The turkey was selected for these studies because turkey skeletal muscle fibroblasts do not grow in primary muscle cell cultures. Results from the proliferation analysis showed a decrease in proliferation by 8 wk of age. Differentiation into myotubes was significantly decreased by 4 wk of age and myotube diameter was decreased. The changes in muscle weight relative to total BW were measured for the anterior latissirnus dorsi, biceps brachii, pectoralis major, sartorius, biceps femoris, and gastrodnemius muscles to compare the relative growth of different muscles. The age at which the muscles reached their maximum relative weight was muscle-dependent, with the biceps brachii plateauing the earliest at 4 wk and the sartorius the latest at 45 wk of age. These data suggested that changes in inyogenic cells begin to occur early in muscle development and the activity of the satellite cells during these initial stages of posthatch growth is critical in overall muscle mass accumulation. [ABSTRACT FROM AUTHOR]

The membrane-associated heparan sulfate proteoglycan families, consisting of the syndecans and glypicans, are low-affinity receptors for fibroblast growth factor 2 (FGF2) that are essential in regulating the cellular response to FGF2. Fibroblast growth factor 2 is a potent stimulator of skeletal muscle cell proliferation and a strong inhibitor of differentiation. The regulation of the expression of the syndecans and glypicans will likely play a role in modulating the effects of FGF2 on cellular growth properties. In the present study, the effect of FGF2 on the expression of syndecan-4 and glypican-1 was measured by real-time PCR during turkey myogenic satellite cell proliferation and differentiation in vitro. Both syndecan-4 and glypican-1 transcription were influenced by the addition of exogenous FGF2. Syndecan-4 mRNA expression was reduced only during proliferation, whereas glypican-i expression was reduced during both proliferation and differentiation. These results suggest that FGF2 growth factor signaling is, in part, regulated by an autoregulatory loop involving FGF2 regulation of syndecan-4 and glypican-1 expression and will affect the growth of skeletal muscle by modulating the proliferation and differentiation of satellite cells. [ABSTRACT FROM AUTHOR]

Pouchitis after total rectocolectomy is the most common complication of ulcerative colitis (UC). The immunological mechanisms involved in the genesis of pouchitis are unclear. Therefore, we evaluated the inflammatory activity in normal ileal pouch mucosa by determining signal transducers and activators of transcription (STAT-1) activation and cytokine expression in patients operated for UC and familial adenomatous polyposis (FAP). Eighteen asymptomatic patients, who underwent total rectocolectomy and J pouch, were evaluated: nine with UC and nine with FAP. The activation of STAT-1 and cytokine expression were determined by immunoblot of total protein extracts from pouch mucosal biopsies. The absence of pouchitis was assessed by clinical, histological and endoscopic parameters, according to the Pouchitis Disease Activity Index. The patients were not receiving any medication. Analysis of variance (anova) and Tukey-Kramer's test were applied. The local ethical committee approved the study and informed consent was signed by all participants. STAT-1 activation was increased in UC when compared to FAP and controls (P < 0.05). Higher levels of interferon (IFN)-gamma expression were observed in UC patients when compared to the control group (P < 0.05), but were similar to FAP. In contrast, cytokine signalling (SOCS-3) and interleukin (IL)-10 expression were similar in all groups (P > 0.05). These findings could explain the higher susceptibility to this inflammatory complication in UC when compared to FAP. A tendency towards increased levels of IFN-gamma and STAT-1 in patients with UC, even without clinical and endoscopic evidence of pouchitis, was observed; studying inflammatory activity in asymptomatic ileal pouches may help understanding of the pathogenesis of pouchitis.

Posthatch muscle growth is determined by the activation, differentiation, and fusion of satellite cells. Satellite cells composing an individual muscle are heterogeneous, which will differentially affect muscle growth. The proliferation and differentiation of turkey primary pectoralis major muscle cells were investigated in vitro at 1 d of age and at 4, 8, 16, 26, 35, 45, and 54 wk of age. The turkey was selected for these studies because turkey skeletal muscle fibroblasts do not grow in primary muscle cell cultures. Results from the proliferation analysis showed a decrease in proliferation by 8 wk of age. Differentiation into myotubes was significantly decreased by 4 wk of age and myotube diameter was decreased. The changes in muscle weight relative to total BW were measured for the anterior latissimus dorsi, biceps brachii, pectoralis major, sartorius, biceps femoris, and gastrocnemius muscles to compare the relative growth of different muscles. The age at which the muscles reached their maximum relative weight was muscle-dependent, with the biceps brachii plateauing the earliest at 4 wk and the sartorius the latest at 45 wk of age. These data suggested that changes in myogenic cells begin to occur early in muscle development and the activity of the satellite cells during these initial stages of posthatch growth is critical in overall muscle mass accumulation.

The occurrence of de novo malignant neoplasias has been shown in postransplant patients under imunosuppression. It is the second leading cause of late death in liver transplant recipients. The greatest incidence is seen in cancers associated with chronic infection by human papilloma virus, skin cancers, oropharyngeal, and gastrointestinal (GI) malignancies. GI stromal tumors (GISTs) are the most common mesenchymal tumors of the GI tract. Rare cases are identified outside the GI tract are collectively known as extragastrointestinal stromal tumors (EGISTs). We present an EGIST case in a liver transplantation patient. A 64-year-old man underwent liver transplantation because of cirrhosis (hepatitis B virus and alcoholism) and hepatocellular carcinoma. Histopathologic findings revealed 2 trabecular hepatocellular carcinomas: a 3.5-cm-diameter lesion located at segment VIII and another 2-cm one at segment V. Seven months later, he noticed a hardened, mobile, painless, 3-cm subcutaneous nodule in the perineum localized in the right lateral quadrant 2 cm distant from the anus. A surgical resection with 1 cm margin yielded a histopathology report of a 5.0 x 3.0 cm spindle cell stromal tumor. The immunohistochemical profile was compatible with a GIST, with 5 mitosis per 50 high-powered fields. This tumor is extremely rare after liver transplantation but has shown a good outcome up to now.

The membrane-associated heparan sulfate proteoglycan families, consisting of the syndecans and glypicans, are low-affinity receptors for fibroblast growth factor 2 (FGF2) that are essential in regulating the cellular response to FGF2. Fibroblast growth factor 2 is a potent stimulator of skeletal muscle cell proliferation and a strong inhibitor of differentiation. The regulation of the expression of the syndecans and glypicans will likely play a role in modulating the effects of FGF2 on cellular growth properties. In the present study, the effect of FGF2 on the expression of syndecan-4 and glypican-1 was measured by real-time PCR during turkey myogenic satellite cell proliferation and differentiation in vitro. Both syndecan-4 and glypican-1 transcription were influenced by the addition of exogenous FGF2. Syndecan-4 mRNA expression was reduced only during proliferation, whereas glypican-1 expression was reduced during both proliferation and differentiation. These results suggest that FGF2 growth factor signaling is, in part, regulated by an autoregulatory loop involving FGF2 regulation of syndecan-4 and glypican-1 expression and will affect the growth of skeletal muscle by modulating the proliferation and differentiation of satellite cells.

The heparan sulfate proteoglycans, syndecan-1 and glypican, are low-affinity receptors for fibroblast growth factor 2 (FGF2). Because FGF2 is a potent stimulator of skeletal muscle cell proliferation and a strong inhibitor of differentiation, it is likely that changes in syndecan-1 and glypican expression will affect myogenesis as both, in part, regulate FGF-dependent signaling. In the current study, expression vector constructs containing either syndecan-1 or glypican were transfected into turkey myogenic satellite cells resulting in the overexpression of these genes. The amount of expression of each of these genes was measured by semiquantitative reverse transcriptase polymerase chain reaction. The satellite cell cultures overexpressing syndecan-1 were unable to fuse to form multinucleated myotubes after differentiation was induced. The syndecan-1-transfected cells maintained a rounded morphology typical of cells during proliferation. In contrast, the satellite cells transfected with glypican formed larger myotubes. These results suggest that both syndecan-1 and glypican play pivotal, but different, roles in both muscle cell proliferation and differentiation.

The low score normal (LSN) chicken exhibits a genetic muscle weakness and altered in vitro myogenesis compared to the normal White Leghorn chicken. The ventricular myosin heavy chain isoform has been reported to be the initial muscle-specific contractile protein expressed during myogenesis. The goals of this study were to determine whether altered myogenesis of the LSN satellite cells in culture was accompanied by delayed ventricular myosin heavy chain expression and to further characterize the altered myogenic events exhibited by the LSN chicken. Immunocytochemical and ELISA analyses were employed to document the temporal expression of the ventricular myosin heavy chain during LSN chicken myogenesis. Satellite cells derived from the LSN chicken pectoralis major exhibited lower (P

The inheritance of, and effect of selection for increased BW on, measurements of muscle fibers and extracellular space in turkeys were studied using a randombred control line (RBC2), a subline (F) of RBC2 selected long-term only for increased 16 wk BW, a commercial sire line (B), and reciprocal crosses of the F and B lines. Measures of additive genetic variation were obtained by comparing all of the pure lines or just the large-bodied F and B lines. Estimates of nonadditive genetic variation were obtained by contrasting the average of the reciprocal crosses with the average of the parental lines. A contrast of the reciprocal crosses provided estimates of sex linkage or maternal effects. Samples of pectoralis major muscle were obtained from three males and three females of each genetic group at 1, 4, 8, and 16 wk of age in a manner to avoid muscle contraction. After fixing and cross sectioning, the muscle samples were stained with hematoxylin and eosin to view muscle morphology. The stained sections were analyzed for muscle fiber width, muscle fiber bundle width (except at 16 wk of age), number of fibers within a 136-microm2 area, and extracellular matrix perimysial (PW) and endomysial (EW) width. Additive genetic variation, as measured by line differences, of measures of muscle fibers and extracellular matrix was a more important source of variation when the RBC2 line was included in the comparison. When all of the pure lines were compared, line differences were significant for fiber bundle width at 4 wk of age; individual fiber width and number of fibers in a given area at 4, 8, and 16 wk of age; PW at all ages; and EW at 1, 8, and 16 wk of age. With the possible exception of PW, nonadditive genetic variation was not an important source of variation for muscle measurements. For PW, the estimates of heterosis were -14.6, 26.4, 14.5, and 17.3% at 1, 4, 8, and 16 wk of age, respectively, but none of the values was significant (P > 0.05). Genetic increases in BW were associated with an increase in muscle fiber width, a smaller number of fibers in a given area, and less extracellular space at older ages. Apparent differences in growth patterns among the genetic groups may have been responsible for the different patterns of change in muscle measurements in the various genetic groups over ages.

Damage to the turkey pectoralis major muscle was studied in a randombred control line (RBC2), a subline (F) from the RBC2 line selected long-term for increased 16-wk BW only, and a commercial sire line (B) at 25 d of incubation and at 1, 4, 8, 16, and 20 wk posthatch. Pectoralis major muscle samples were obtained from three females and three males from each line in a manner to avoid contraction. After being fixed and sectioned, the muscle samples were stained with hematoxylin and eosin to view muscle fiber and muscle fiber bundle morphology. Beginning at 8 wk posthatch, differences in muscle fiber morphology were observed among the different lines. The RBC2 line throughout the duration of the study maintained well-organized muscle fibers and muscle fiber bundles with large capillary networks. In contrast, the growth-selected F line began to show muscle fiber degeneration at 8 wk posthatch, and limited capillary beds were observed as development proceeded. The B line had intermediate muscle morphology between the RBC2 and F lines, but by 20 wk posthatch significant muscle fiber degeneration was present with limited capillary supply. The degenerative muscle fiber changes were predominant in the growth-selected F-line, suggesting that growth selection for just BW may be associated with posthatch muscle damage.

Muscle development at 20 and 25 d of incubation was studied in a randombred control line (RBC2), a subline (F) of RBC2 selected only for increased 16-wk BW, a commercial sire line (B), and reciprocal crosses of the F and B lines. Muscle samples from three males and three females of each genetic group were collected in such a manner to avoid contraction. After fixing, the muscles were stained with hematoxylin and eosin, measurements of muscle fiber width, muscle fiber bundle length and width, number of fibers within a 15.6 microm2 area, and extracellular matrix perimysial (PW) and endomysial (EW) width were taken with an Olympus XI 70 microscope equipped with an Olympus Magna Fire digital camera linked to Image Pro software. From each slide, 20 measurements were taken for each characteristic analyzed. In most of the muscle traits measured, additive genetic variation, as indicated by line differences, occurred when the RBC2 line was included in the comparison of pure lines. However, when only the B and F lines were compared, line differences were less frequent. In comparisons of the B and F lines and their reciprocal crosses, heterosis, as measured by contrasts of the average of the pure lines and the average of the reciprocal crosses, was an important source of variation for individual fiber measurements (negative) and extracellular space (positive) at 20 d of incubation but was less important at 25 d of incubation. No significant interactions between genetic group and sex were noted at 20 d of incubation, but such interactions were frequent at 25 d of incubation. These results suggest that muscle organizational differences between the two sexes begin to occur between these two ages and are not the same for different genetic groups.

Skeletal muscle development is, in part, regulated by myoblast-extracellular matrix interactions mediated by the transmembrane integrin family of heterodimeric receptors. The avian genetic muscle weakness, low score normal (LSN), exhibits modified myotube and sarcomere structure that may be associated with altered integrin expression. Protein expression of the beta1 integrin subunit was measured during normal and LSN Pectoralis major muscle development at 14, 16, and 18 d of embryonic development, and 1 d and 1 and 6 wk posthatch. During embryonic development, integrin expression was downregulated. However, by 1 wk posthatch, integrin levels were upregulated and remained elevated through 6 wk posthatch. This pattern was observed in both normal and LSN muscle development. Overall, beta1 integrin levels were lower in the LSN P. major muscle. In normal and LSN satellite cell cultures, beta1 integrin expression was low during proliferation. In early differentiation, beta1 integrin expression increased and was then downregulated. As observed in the muscle extracts, LSN beta1 integrin expression was significantly lower during differentiation. These results suggest that the regulation of beta1 integrin expression is critical to the progression of myogenesis, and, during LSN myogenesis, decreased expression of beta1 integrin may be associated with modifications in muscle structure.

The avian Low Score Normal (LSN) genetic muscle weakness is phenotypically characterized by a reduction in the ability of birds to right themselves from a supine position. In previous studies, LSN birds exhibited elevated levels of decorin transcript and protein at embryonic Day 20 and a large increase in collagen crosslinking at 6 wk posthatch. Transforming growth factor-beta (TGF-beta) expression has been reported to control decorin expression. Steady state levels of TGF-beta1 and TGF-beta2 transcripts were determined in ovo and posthatch in order to initiate an investigation of the mechanism underlying decorin expression. On embryonic Day 20 through 1 wk posthatch, TGF-beta2 transcript levels were elevated, whereas an increase in TGF-beta1 was only noted at 1 d posthatch. These data suggest that the increase in decorin expression may be associated with a modification in a TGF-beta signal transduction pathway.

The avian Low Score Normal (LSN) genetic muscle weakness is phenotypically characterized by a reduction in the ability of the birds to right themselves from a supine position. Compared to normal skeletal muscle, LSN muscle has normal myosin isoform switching and cell-cell recognition, elevated glycosaminoglycan and decorin levels at embryonic Day 20, and a large increase in collagen crosslinking at 6 wk posthatch. To begin to determine the biological mechanism involved in the elevated decorin protein concentration at embryonic Day 20, the steady-state levels of transcripts encoding both decorin and collagen Type I at embryonic Days 14, 19, and 20, and at 1 d and 6 wk posthatch were measured. On embryonic Day 20, collagen Type I transcripts were not different from the control but there was a significant elevation in decorin transcript levels. At 1 d and 6 wk posthatch, transcript levels of decorin and collagen Type I were not different between LSN and controls. The change in decorin transcript steady-state levels is limited to late embryonic development and suggests an alteration in a signal transduction pathway regulating decorin transcription.

Purpose: Duodenal compression by the superior mesenteric artery following total proctocolectomy and ileal pouch-anal anastomosis is a rare occurrence. Previous surgical treatment involved duodenal division. The aim of this report was to describe a case with such a complication and to discuss an operative alternative., Methods: Case report., Results: Mobilization of the duodenum from its retroperitoneal attachments, without transection and reanastomosis, allowed the free passage of gas through the duodenum and recovery for the patient., Conclusion: This case report suggests that a more conservative approach may be successful in managing this complication.

A new ileal pouch design, combining an upper triplicate ileum with a lower duplicate ileum, is described. The physical characteristics of this two-chamber reservoir would lead to better functional results by delaying the filling time of the reservoir.

The etiological diagnosis for the colorectal strictures can be very difficult. With the colonoscopy, it was possible to obtain samples with direct vision of the lesion. Consequently, the cytologic efficiency improved, with similar or even better results than histopathology. In the present study (107 patients) it was used the cytopathology performed during colonoscopy. After a specimen was obtained, it was smeared on glass slide, fixed in 95% alcohol, stained by HE, and examined microscopically, with conclusions during the procedure. The results were compared and further correlated to the histopathology of surgical specimen or based on clinical follow-up in those case not submitted to operation. The results showed a striking degree of correlation between both exams. For the cytopathology there were no inconclusive results, false-negative were less frequent than with histopathology, and only one case of false-positive (rectal villus adenoma) was observed. Considering the type, localization and perviousness of the lesion, similar conclusions were drawn, and did not affect the results, except that there were more false-negatives in both exams when the lesions were impervious. The cytopathological and histopathological studies were both highly sensitive and specific for the diagnosis of carcinoma. In conclusion, cytopathology performed during colonoscopy is safe and efficient in the study of colorectal strictures, which contributes to improve the orientation of these patients.