Engineering of pre‐vascularized urethral patch with muscle flaps and hypoxia‐activated hUCMSCs improves its therapeutic outcome

Tissue engineering has brought new hopes for urethral reconstruction. However, the absence of pre‐vascularization and the subsequent degradation of materials often lead to the failure of in vivo application. In this study, with the assistance of hypoxia‐activated human umbilical cord mesenchymal stem cells (hUCMSCs), pedicled muscle flaps were used as materials and pre‐incubated in ventral penile subcutaneous cavity of rabbit for 3 weeks to prepare a pre‐vascularized urethral construct. We found that small vessels and muscle fibres were scattered in the construct after 3 weeks' pre‐incubation. The construct presented a fibrous reticular structure, which was similar to that of the corpus spongiosum under microscope examination. The produced constructs were then used as a patch graft for reconstruction of the defective rabbit urethra (experimental group), natural muscular patch was used as control (control group). Twelve weeks after the reconstructive surgery, urethrography and urethroscope inspections showed wide calibres of the reconstructed urethra in the experimental group. Histopathological studies revealed that fibrous connective tissues and abundant muscle fibres constituted the main body of the patch‐grafted urethra. In contrast, in the control group, only adipose tissue was found in the stenosis‐reconstructed urethra, replacing the originally grafted muscular tissue. To our knowledge, this is the first report that successfully constructed a pre‐vascularized urethral construct by using hypoxia‐activated hUCMSC and pedicled muscle flaps. More importantly, the pre‐vascularized construct showed a good performance in urethral reconstruction when applied in vivo. The study provided a novel strategy for tissue engineering of pre‐vascularized urethral construct for the defective urethra, representing a further advancement in urethral reconstruction.     

Vestibular, vaginal and urethral relationships in spayed and intact normal dogs

The purpose of this study was to evaluate the urogenital anatomy in sound spayed and intact dogs. Fourteen intact and 12 spayed sound dogs had their lower urogenital tract evaluated with conventional vaginourethrography, computed tomography (CT) vaginourethrography and uroendoscopy under general anesthesia. Measurements for each of the three imaging modalities were made. Measurements included vaginal length, vaginal height, vaginal width, vestibule length, vestibule height, vestibule width, cingulum height, cingulum width, cingulum area, urethral length, urethral height, urethral width, angle of urethra to vestibule and angle of vagina to vestibule. Group comparisons were made between sound spayed and intact dogs. In general, most measurements were greater in intact dogs compared to spayed dogs regardless of imaging modality utilized. Group-weight interactions (P<0.01) were found on conventional vaginourethrograms (vaginal height and length and vestibule height, length and width) and CT vaginourethrograms (vaginal length, vestibule height, length and width, cingulum area, urethral width and angle of vagina to vestibule). A three-way interaction (P<0.01) for vaginal length, vaginal height and vestibule height was also identified. Vestibule length and height, cingulum height and urethral length were larger (P<0.01) with increasing dog weight and in intact dogs for both conventional and CT vaginourethrograms. No difference in vestibulovaginal ratio was observed between intact dogs and spayed dogs (P=0.0221). One of 14 intact dogs and 8 of 12 spayed dogs had vestibulovaginal ratios <0.33 on conventional vaginourethrograms. Anatomical differences in the lower urogenital tract between spayed and intact dogs emphasizes the need to establish normal reference ranges for spayed and intact dogs.     

Single-surgeon experience of 87 buccal mucosal graft harvests

Although buccal mucosa has been the preferred graft material for use in the reconstruction of anterior urethral strictures for approximately 20 years, the plastic surgery literature is lacking in studies describing and evaluating this technique. The authors sought to report their experience with the technique of buccal mucosa graft harvest for use in urethral reconstruction. The authors identified 87 consecutive patients who underwent buccal mucosal graft harvesting during a 15-year period from 1995 to 2010. All donor sites were closed primarily. Preoperatively, a specialty-specific evaluation was performed by plastic surgeons and urologists. Postoperative donor-site morbidity was followed clinically. Eighty-seven patients underwent buccal harvesting. A single buccal graft was harvested in 75 patients (86.2 percent) and bilateral grafts were harvested in 12 patients (13.8 percent). The median patient age of the patients was 42 years (range, 16 to 78 years). There were no major intraoperative or postoperative complications. Minor complications included mild discomfort, numbness, cheek swelling, and restriction of oral movement, all of which resolved by the 3-month postoperative visit. The ability to eat and drink postoperatively was diminished during only the first 24 to 48 hours in all patients. Our series provides a 1-year follow-up in a substantial cohort looking at clinical outcomes after buccal harvesting. These results confirm the safety and efficacy of buccal mucosal harvesting to provide an effective and cosmetically superior option for urethral reconstruction. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, IV.     

Expression of the androgen receptor and 5α-reductase type 2 in the developing human fetal penis and urethra

Normal penile development is dependent on testosterone, its conversion via steroid 5 alpha-reductase type 2 to dihydrotestosterone, and a functional androgen receptor (AR). The goal of this study was to investigate the distribution of AR and 5 alpha-reductase type 2 in the developing human fetal external genitalia with special emphasis on urethra formation. Twenty fetal genital specimens from normal human males (12-20 weeks gestation) were sectioned serially and stained by avidin-biotinylated peroxidase complex method with antigen retrieval. Stained sections throughout male genital development documented the expression of AR and 5 alpha-reductase type 2 in the phallus. Between 12 and 14 weeks of gestation, AR was localized to epithelial cells of the urethral plate in the glans, the tubular urethra of the penile shaft, and stromal tissue surrounding the urethral epithelium. In the fetal penis between 16 and 20 weeks gestation, the density of AR expression was greatest in urethral epithelial cells versus the surrounding stromal tissues. There was a characteristic pattern of AR expression in the glandular urethral epithelium between 16 and 20 weeks gestation. AR expression was greater along the ventral aspect of the glandular urethra than along the dorsal aspect of the urethral epithelium. The expression of 5 alpha-reductase type 2 was localized to the stroma surrounding the urethra, especially along the urethral seam area in the ventral portion of the remodeling urethra. These anatomical studies support the hypothesis that androgens are essential for the formation of the ventral portion of the urethra and that abnormalities in either the AR or 5 alpha-reductase type 2 can explain the occurrence of hypospadias.     

New surgical technique to correct urovagina improves the fertility of dairy cows

Several surgical techniques have been proposed for the treatment of urovagina as a major cause of infertility in cows. However, so far no technique seems to be successful in all cases. Given that an incompetent or damaged constrictor vestibuli muscle is a feature of cows with urovagina, we hypothesized that surgical correction of urovagina by cerclage of the vestibulovaginal junction under the vaginal wall cranial to the urethral opening would prevent cranial flow of urine and improve fertility. Our study was performed on 39 non-pregnant lactating Holstein-Friesian cows suffering urovagina, with a vaginal content of urine exceeding 100mL and with evident incompetence of the constrictor vestibuli muscle. Cows were randomly assigned to a Control (untreated cows, n=20) or Experimental (n=19) group. An encircling polydioxanone suture was placed in the vaginal wall at the vestibulovaginal junction to create a vestibulovaginal cerclage in the Experimental cows. Surgery was observed to resolve urovagina in 17 (89.5%) of the 19 treated cows. Pregnancy was recorded in 7/20 (35%) and 14/19 (74%) cows in the Control and Experimental groups, respectively. Using logistic regression procedures and based on the odds ratio, we determined that cows undergoing surgical correction of urovagina were 5.2 times more likely to become pregnant than untreated cows (P=0.015). Our results suggest that vestibulovaginal junction cerclage prevents the cranial flow of urine and improves the function of the constrictor vestibuli muscle in cows suffering urovagina. They also indicate that, under these conditions, urovagina correction may dramatically increase fertility.     

Effect of epidermal growth factor on intestinal adaptation after allogeneic small bowel transplantation in rats

We reported that epidermal growth factor (EGF) stimulated graft adaptation in a rat model of syngeneic small bowel transplantation. However, graft rejection is a severe problem with clinical small bowel transplantation, because small intestinal wall contains large amounts of lymphoid tissue. Studies were performed to investigate the effect of EGF on allogeneic graft adaptation after small bowel transplantation in rats treated with an immunosuppressant FK506. The transplanted animals received intraperitoneally EGF or saline (untreated) after surgery and were examined for analysis one week later. EGF-treated group markedly enhanced the water absorption and induction of sodium glucose cotransporter (SGLTI) as compared with EGF-untreated group. EGF-treated group also increased the mucosal crypt depth and its cell proliferating rate, although there was no significant difference in the mucosal villus height between the two groups. These results indicate that EGF accelerates intestinal allograft adaptation in part by the recovery of mucosal structure and function after small bowel transplantation in rats. EGF may have relevance to promote graft function in clinical small intestinal transplantation.     

Ontogeny of the male urethra: theory of endodermal differentiation

The most widely accepted mechanism of male urethral development proposes that the urethral plate is elevated by urethral folds which fuse ventrally in a proximal-to-distal sequence. Unlike its proximal counterpart, the urethra which forms within the glans is lined by a stratified squamous epithelium and has a more controversial development. One theory supports the idea that fusion of the urethral folds extends all the way to the tip of the glans. Another theory suggests that a solid ectodermal in-growth of epidermis canalizes the glandar urethra. We hypothesized that the use of immunohistochemical staining and tissue recombinant grafting would delineate the epithelia involved and lend clues to their origin. Thirty-six human fetal phallic specimens of gestational ages 5-22 weeks were sectioned and stained immunohistochemically with antibodies raised against different cytokeratins. Evaluation of the sections showed that the urethral plate, an extension of the urogenital sinus, extended to the tip of the phallus and maintained patency and continuity throughout the process of urethral development. The entire urethra, including the glans portion, was formed by dorsal extension and disintegration of the urethral plate combined with ventral growth and fusion of the urethral folds. Sections of the distal glandar urethra showed no evidence of a solid ectodermal ingrowth. Rather, immunostaining results at different ages suggested differentiation of the endodermal urethral plate into a stratified squamous epithelium. To determine whether urothelium could be induced to express a stratified squamous phenotype, mouse fetal bladder epithelium was combined with rat fetal genital tubercle mesenchyme and grown under the renal capsule of athymic mice. The bladder epithelium differentiated into a stratified squamous epithelium. Thus, proper mesenchymal signaling may induce differentiation of urothelium into a stratified squamous phenotype, such as during development of the urethra of the glans penis.     

Effects of bull exposure on the cyclic activity of beef cows

In Experiment I, 38 crossbred suckled beef cows grazing fescue pastures and 34 crossbred beef cows grazing bluestem pastures were randomly allocated at the time of calving into a group with 4 teaser bulls or no bulls. Two blood samples were collected 7 d apart from the cows to determine cyclic activity 67 and 76 d after calving in the fescue and bluestem pastures, respectively. Progesterone greater than 1.0 ng/ml in one or both samples indicated cyclic activity. There was no difference in the percentage of cows cyclic among the different groups. The number of cyclic cows in the fescue pasture with bulls was 16/19 (84%); in the fescue pasture with no bulls, 14/19 (74%); in the bluestem pasture with bulls, 17/17 (100%); and in the bluestem pasture with no bulls, 16/17 (94%). Overall cyclic activity among all cows for teaser bull-exposed and no bull was similar, 33/36 (91%) and 30/36 (83%). Overall cyclic activity was greater (P < 0.05) in cows grazing bluestem (33/34), 97% than fescue pastures (30/38), 80%. Measurements of cyclic activity were initiated too late in the postcalving period to quantify differences in estrous activity between the bull and no bull treatment groups. Another trial was planned for the following year with a modified protocol. In Experiment II, blood samples were collected for progesterone concentrations soon after calving and were repeated at intervals to characterize both the occurrence and duration of estrous cycles. In this experiment, 29 crossbred suckled beef cows grazing fescue pastures were randomly allocated 12 d after calving (Day 0) into 1 of 2 groups with teaser bulls or without bulls. Nineteen crossbred beef cows grazing bluestem pastures were allocated similarly 10 d after calving (Day 0). Bulls were added to the groups with bulls in fescue and bluestem pastures on day 6 after the initial allocations. Blood samples were collected from all cows on Day 0 and every 3 d until Day 46. Means (+/- SEM) of the cumulative progesterone concentrations (ng/ml) per cow for the 16 samples from cows grazing fescue were 12.5 +/- 3.5 for cows exposed to bulls, 2.5 +/- 0.16 for cows not exposed to bulls, 27.6 +/- 4.42 for cows grazing bluestem pastures and exposed to bulls, and 16.0 +/- 2.75 for cows without exposure to bulls. Progesterone concentrations were higher in cows exposed to bulls (P < 0.01). The percentages of both short and normal cycles increased (P < 0.01) in groups exposed to bulls (88%, 21/24 and 63%, 15/24) when compared with the no bull groups (29%, 7/24 and 21%, 5/24), respectively. Cows exposed to bulls also showed increased cyclic activity.     

Biostimulatory effects of estrous cows and bulls on resumption of ovarian activity in postpartum anestrous Zebu (Bos indicus) cows in the humid tropics

Suckled postpartum Zebu (Bos indicus) cows were exposed either to estrous females alone or together with bulls to determine if this regimen would stimulate the resumption of ovarian activity. The study was carried out on 60 Zebu cows. The animals were allocated at parturition to 1 of 3 treatment groups. Group FB cows (n = 14) were exposed to progestogen treated females (approximately 3 cows per progestogen treated cow) and a bull for 42 d. Group F cows (n = 31) were exposed to progestogen treated females and after 21 d a bull was introduced and remained with the group until the end of the experiment 21 d later. Estrus was induced with Syncro Mate B treatment and the cows were in estrus when introduced into Groups FB and F. Group N cows (n = 15) were introduced to a bull 21 d after the treatments had begun in Groups FB and F, and the bull remained with these cows until the end of the study. On average, resumption of ovarian activity occurred in 57% (8/14), 19% (6/31) and 7% (1/15) of the cows in Group FB, F and N, respectively, within 21 d after the start of treatments. The difference was significant between Group FB and Groups F and N (P < 0.05). During the second part of the experiment, in which the bull was introduced to Groups F and N, an additional 32% (10/31) of the cows in Group F and 33% (5/15) in Group N resumed ovarian activity. No cows in Group FB resumed ovarian activity during this period. There were significant differences in body condition between cows that resumed ovarian activity and those that remained in anestrus (P < 0.05). We conclude that biostimulation effectively enhanced resumption of ovarian activity in anestrous suckled Zebu cows. This technique offers a potentially useful and practical way to improve reproductive efficiency in anestrous Zebu cattle in the tropics.     

Successful replantation in the face of absent venous drainage: an experimental study

Inadequate venous outflow is the factor most responsible in digital replantation failure and is a common cause of tissue loss in general. An experimental replantation model utilizing the rabbit ear was used to study the extreme example of venous congestion: absent venous drainage in the replanted part. Results of this study support the use of single arterial inflow along with an efferent AV fistula for outflow in the management of replants with absent venous drainage. Potential indications for the use of an efferent arteriovenous fistula in digital revascularization include the following: (1) the finding of distal veins too small to reanastomose, as is often the case in children and at distal levels in adults; (2) preferential destruction of distal venous structures, as commonly seen in degloving or other dorsal injuries; and (3) in the management of postreplant venous thrombosis.     

Relocations of cell convergence sites and formation of pharyngula-like shapes in mechanically relaxed <Emphasis Type="Italic">Xenopus</Emphasis> embryos

Influence of the relaxation of mechanical tensions upon collective cell movements, shape formation, and expression patterns of tissue-specific genes has been studied in Xenopus laevis embryos. We show that the local relaxation of tensile stresses within the suprablastoporal area (SBA) performed at the early-midgastrula stage leads to a complete arrest of normal convergent cell intercalation towards the dorsal midline. As a result, SBA either remains nondeformed or protrudes a strip of cells migrating ventralwards along one of the lateral lips of the opened blastopore. Already, few minutes later, the tissues in the ventral lip vicinity undergo abnormal transversal contraction/longitudinal extension resulting in the abnormal cell convergence toward ventral (rather than dorsal) embryo midline. Within a day, the dorsally relaxed embryos acquire pharyngula-like shapes and often possess tail-like protrusions. Their antero-posterior and dorso-ventral polarity, as well as expression patterns of pan-neural (Sox3), muscular cardiac actin, and forebrain (Otx2) genes substantially deviate from the normal ones. We suggest that normal gastrulation is permanently controlled by mechanical stresses within the blastopore circumference. The role of tissue tensions in regulating collective cell movements and creating pharyngula-like shapes are discussed.     

The muscle layer of the canine gallbladder and cystic duct

The muscle layer of the canine gallbladder wall and cystic duct was found to be a three-dimensional meshwork of smooth muscle bundles which appear loosely and irregularly arranged on the mucosal aspect and consolidate to form a homogeneous plate-like layer on the serosal aspect. The muscle bundles are tightly woven around interspersed pockets of loose connective tissue in the gallbladder wall and gradually become loosely arranged with more prominent amounts of intervening connective tissue in the cystic duct. The muscle layer is thickest in the gallbladder wall and becomes progressively thinner out into the cystic duct. No anatomic sphincter was observed. Ultrastructural organization revealed individual muscle fibers to be of irregular profile, often branching, widely spaced with intervening collagen fibers, and having few cell-to-cell contacts.     

女性尿道阴道瘘数字化三维重建在临床治疗中的初步应用

目的:基于CT扫描图象建立精确的女性尿道阴道瘘数字化模型,探讨其在临床诊断及治疗中的应用。方法:选择1例女性尿道阴道瘘病例,进行尿道CT连续断层扫描,扫描结果导入Mimics软件中进行三维重建,利用三维重建模型指导临床。结果:建立女性尿道阴道瘘及周围结构的三维立体模型,可以方便地从任意角度和方向观察瘘管情况,测量有关的数据;还可以在数字化模型上进行手术设计。结论:女性尿道阴道瘘的数字化三维模型能够更直观、准确地反映病变部位的三维立体结构。对女性尿道阴道瘘的诊断、手术规划等有较大帮助。     

Changes in vulvar and vestibular tissue of the bovine during the estrous cycle as determined by the use of near-infrared interactance

Near-infrared interactance spectra of the vulva and vestibule of cows were analyzed throughout the estrous cycle to determine if they could be used to detect tissue changes that are associated with estrus. The spectra were collected daily from 16 cyclic Holstein cows beginning at 40 days post partum and continuing for 3 estrous cycles, or until pregnancy was verified. Spectra collected on the day when estrus was exhibited were compared with spectra collected on the nonestrous days. There were spectral differences at 1695 to 1705, at 1790 to 1800 and at 1880 to 1900 nm (P<0.05), regions that are associated with changes in carbohydrate, protein, and water content of the tissues, respectively. Discriminant analysis of interactance measurements at the significant wavelengths was used to develop an equation to predict the day of estrus. The error rate associated with the equation was 26%. Results indicate that near-infrared analysis can detect changes in vestibular and vulvar tissue during the estrous cycle. However, accurate estrus detection using near-infrared interactance techniques will require additional study to reduce variation in the results.     

Bull effect on the reproductive performance of mature and first calf-suckled Zebu cows in the tropics

This experiment was designed to determine if continuous bull exposure of heifers 2 wk post partum to a first calf (30 to 36 m old) or mature (42 to 72 m old) Zebú Brahman suckled cows reduces the interval to first estrus and the number of days open. Post partum first calf cows were randomly assigned to be exposed continuously to epidectomized bulls with surgical ablation of the dorsal ligament of the penis (Group 35 FBE, n = 25) or not exposed to bulls (Group FNE, n = 22). Post partum mature cows were likewise assigned to be continuously exposed (Group MBE, n = 24) or not exposed to bulls (Group MNE, n = 25). Milk samples were collected twice a week for progesterone assays to determine resumption of ovarian cyclic activity. All the cows were artificially inseminated for 120 d while in their respective treatment groups. The first postpartum estrus percentages were 60 < 90 d, 24.4%; and > 90 d, 33.7%. The interval to first estrus f-or Groups FBE and FNE was 75.4 and 104.2 d, respectively, and differed significantly (P < 0.01). For Groups MBE and MNE the interval to first estrus was 76.3 and 95.0 d, respectively (P < 0.05). The number of open days for Groups FBE and FNE was 77.3 and 114.9, respectively (P < 0.01); while for Groups MBE and MNE it was 73.0 and 98.6 d, respectively (P < 0.01). The results of the present study indicate that introduction of bulls to first calving cows or mature Zebú Brahman cows after parturition provides a practical and economical option for reducing the extended post partum anestrus interval and days open under tropical conditions.     

Intrauterine infusion of highly enriched bovine trophoblast protein-1 complex exerts an antiluteolytic effect to extend corpus luteum lifespan in cyclic cattle

Intrauterine infusion of enriched bovine trophoblast protein-1 complex (bTP-1) resulted in extension of interoestrous intervals and corpus luteum function in cyclic cattle. Conceptus proteins were obtained by culture of Day 17-18 conceptuses for 72 h. Media from the first (n = 28), second (n = 26) and third (n = 19) 24 h of conceptus incubations were utilized. A highly enriched preparation of bTP-1 was obtained by a combination of ammonium sulphate precipitation, ion-exchange chromatography, and h.p.l.c. gel filtration. Degree of purity of the final preparation was confirmed by gel electrophoresis and immunoblotting with antiserum to ovine trophoblast protein-1. Jersey cattle (3 per group) received intrauterine infusions, twice daily from Day 15.5 to 21.0, of bovine serum albumin, the entire array of bovine conceptus secretory proteins (bCSP) from the 3 days of conceptus culture, or bTP-1. Infusions were via a catheter into the uterine horn ipsilateral to the corpus luteum. Oestrous cycle length in bTP-1-treated cows (26.1 +/- 1.3 days) was greater than for cows given BSA (19.5 +/- 1.3 days) or bCSP (21.5 +/- 1.3 days). Similarly, progesterone concentrations in serum remained elevated for a longer period of time for bTP-1-treated cows than for cows treated with BSA or bCSP. Residual variance associated with vena cava concentrations of PGF-2 alpha at Days 19-21 after oestrus (which included the variance between 15-min periods within cows) was reduced in cows treated with bTP-1 as compared to other groups. Lack of a bCSP effect may have been due to low amounts of bTP-1 in conceptus-conditioned medium from cultures of greater than 24 h. None the less, purified bTP-1 was effective in extending luteal function and appears to be the antiluteolytic agent of early pregnancy.     

Osteocutaneous flap prefabrication based on the principle of vascular induction: an experimental and clinical study

Conventional osteomyocutaneous flaps do not always meet the requirements of a composite defect. A prefabricated composite flap may then be indicated to custom create the flap as dictated by the complex geometry of the defect. The usual method to prefabricate an osteocutaneous flap is to harvest a nonvascularized bone graft and place it into a vascular territory of a soft tissue, such as skin, muscle, or omentum, before its transfer. The basic problem with this method is that the bone graft repair is dependent on the vascular carrier; the bone needs to be revascularized and regenerate. The bone graft may not be adequately perfused at all, even long after the transfer of the prefabricated flap. This study was designed to prefabricate an osteocutaneous flap where simply the bone nourishes the soft tissues, in contrast to the conventional technique in which the soft tissue supplies a bone graft. This technique is based on the principle of vascular induction, where a pedicled bone flap acts as the vascular carrier to neovascularize a skin segment before its transfer. Using a total of 40 New Zealand White rabbits, two groups were constructed as the experimental and control groups. In the experimental group, a pedicled scapular bone flap was induced to neovascularize the dorsal trunk skin by anchoring the bone flap to the partially elevated skin flap with sutures in the first stage. After a period of 4 weeks, the prefabricated composite flaps (n = 25) were harvested as island flaps pedicled on the axillary vessels. In the control group, nonvascularized scapular bone graft was implanted under the dorsal trunk skin with sutures; after 4 weeks, island composite flaps (n = 15) were harvested pedicled on the cutaneous branch of the thoracodorsal vessels. In both groups, viability of the bony and cutaneous components was evaluated by means of direct observation, bone scintigraphy, measurement of bone metabolic activity, microangiography, dye injection study, and histology. Results demonstrated that by direct observation on day 7, the skin island of all of the flaps in the experimental group was totally viable, like the standard axial-pattern flap in the control group. Bone scintigraphy revealed a normal to increased pattern of radionuclide uptake in the experimental group, whereas the bone graft in the control group showed a decreased to normal pattern of radioactivity uptake. The biodistribution studies revealed that the mean radionuclide uptake (percent injected dose of 99mTc methylene diphosphonate/gram tissue) was greater for the experimental group (0.49+/-0.17) than for the control group (0.29+/-0.15). The difference was statistically significant (p<0.01). By microangiography, the cutaneous component of the prefabricated flap of the experimental group was observed to be diffusely neovascularized. Histology demonstrated that although the bone was highly vascular and cellular in the experimental group, examination of the bone grafts in the control group revealed necrotic marrow, empty lacunae, and necrotic cellular debris. Circulation to the bone in the experimental group was also demonstrated by India ink injection studies, which revealed staining within the blood vessels in the bone marrow. Based on this experimental study, a clinical technique was developed in which a pedicled split-inner cortex iliac crest bone flap is elevated and implanted under the medial groin skin in the first stage. After a neovascularization period of 4 weeks, prefabricated composite flap is harvested based on the deep circumflex iliac vessels and transferred to the defect. Using this clinical technique, two cases are presented in which the composite bone and soft-tissue defects were reconstructed with the prefabricated iliac osteomyocutaneous flap. This technique offers the following advantages over the traditional method of osteocutaneous flap prefabrication. Rich vascularity of the bony component of the flap is preserved following transfer (i.e. (ABSTRACT     

The effect of a GnRH analogue on the dynamics of follicular development and synchronization of estrus in lactating cyclic dairy cows

A GnRH analogue was used to synchronize ovarian follicular development prior to an injection of PGF(2alpha) for the synchronization of estrus in lactating Holstein cows. On Day 12 (estrus = Day 0) of the experimental cycle, cows (n = 8) were injected with 8 mug Buserelin (BUS group), followed by 25 mg PGF(2alpha) 7 d later (Day 19). Control cows (n = 7) received PGF(2alpha) on Day 12 (PGF group). Ovaries were scanned daily via ultrasonography, and plasma progesterone and estradiol concentrations were determined. Sizes of all visible follicles were recorded. Follicles were classified as small (3 to 5 mm), medium (6 to 9 mm), or large (>/= 10 mm). Between Days 12 and 16 of the cycle, the number of large follicles in PGF cows remained unchanged (1.2), whereas in the BUS group, the number of large follicles decreased from 1.3 on Day 12 to 0.5 on Day 15. Only 4 of 7 PGF cows ovulated a second-wave dominant follicle. In the BUS group, 7 of 8 cows ovulated a GnRH analogue induced dominant follicle that was first identified on Day 15. During the follicular phase (last 5 d prior to estrus), plasma progesterone declined in association with CL regression in both groups, and estradiol concentrations increased, reaching higher (P<.0.05) preovulatory peak concentration in BUS cows than in PGF cows (14.0 +/- 1.0 vs 10.4 +/- 1.1 pg/ml). The number of medium-size follicles was smaller and the number of small-size follicles tended to be higher in BUS cows than in the PGF-treated group. On the day of estrus, the size of the ovulatory follicle (16.1 vs 13.3 mm) and the size difference between the ovulatory and second largest follicle (11.4 vs 6.2 mm) were both larger in BUS cows than in PGF-treated cows, suggesting a more potent dominance effect of the ovulatory follicle in the BUS cows. This study suggests that a GnRH analogue can alter follicular development prior to synchronization of estrus with an injection of PGF(2alpha) in lactating dairy cows.     

Tissue Engineering for Human Urethral Reconstruction: Systematic Review of Recent Literature

BackgroundTechniques to treat urethral stricture and hypospadias are restricted, as substitution of the unhealthy urethra with tissue from other origins (skin, bladder or buccal mucosa) has some limitations. Therefore, alternative sources of tissue for use in urethral reconstructions are considered, such as ex vivo engineered constructs.PurposeTo review recent literature on tissue engineering for human urethral reconstruction.MethodsA search was made in the PubMed and Embase databases restricted to the last 25 years and the English language.ResultsA total of 45 articles were selected describing the use of tissue engineering in urethral reconstruction. The results are discussed in four groups: autologous cell cultures, matrices/scaffolds, cell-seeded scaffolds, and clinical results of urethral reconstructions using these materials. Different progenitor cells were used, isolated from either urine or adipose tissue, but slightly better results were obtained with in vitro expansion of urothelial cells from bladder washings, tissue biopsies from the bladder (urothelium) or the oral cavity (buccal mucosa). Compared with a synthetic scaffold, a biological scaffold has the advantage of bioactive extracellular matrix proteins on its surface. When applied clinically, a non-seeded matrix only seems suited for use as an onlay graft. When a tubularized substitution is the aim, a cell-seeded construct seems more beneficial.ConclusionsConsiderable experience is available with tissue engineering of urethral tissue in vitro, produced with cells of different origin. Clinical and in vivo experiments show promising results.     

Persistent dominant follicle alters pattern of oviductal secretory proteins from cows at estrus.

The experimental objective was to compare synthesis of oviductal secretory proteins of dairy cows bearing a persistent dominant follicle (PDF) versus a fresh dominant follicle (FDF) at estrus. On Day 7 after synchronized estrus (Day 0), cows received an intravaginal progesterone device and injection of prostaglandin F2alpha (PGF2alpha). On Day 9, cows received an injection of a GnRH agonist (FDF group; n = 3) or received no injection (PDF group, n = 3). On Day 16, all cows received PGF2alpha, and progesterone devices were removed. At slaughter on Day 18 or Day 19, oviducts ipsilateral and contralateral to the dominant follicle were divided into infundibulum, ampulla, and isthmus regions. Explants from oviductal regions were cultured in minimal essential medium supplemented with [3H]leucine for 24 h. Two-dimensional fluorographs of proteins in conditioned media were analyzed by densitometry. Rate of incorporation of [3H]leucine into macromolecules was greater in the infundibulum, ampulla, and isthmus of FDF cows (p < 0.01). Overall, intensities of radiolabeled secretory protein (P) 2 and P13 were greater for FDF than for PDF. In the ampulla, P14 was more intense for FDF while P7 was more intense for PDF. Abundance of P1 in the isthmus was greater for PDF cows. Across regions, P5, P6, P8, P9, and P11 were more intense for PDF than for FDF in the ipsilateral side. In the contralateral side, P19 was more intense for PDF than for FDF, whereas P6, P8, P9, and P11 were more intense for FDF. Differences in biosynthetic activity and in secreted oviductal proteins from cows bearing a PDF may contribute to the decrease in fertility associated with a PDF.