Microvascular breast reconstruction and thromboembolic events in patients on hormone therapy: Audit of practice from a tertiary referral centre.

INTRODUCTION: Hormonal therapy with tamoxifen and aromatase inhibitors reduces breast cancer recurrence and mortality but represents a risk factor for thromboembolic events. Therefore, most surgeons discontinue hormonal agents before microvascular surgery and for a variable period thereafter. There are no guidelines regarding when therapy should be stopped (preoperatively) or when it should be resumed (post-operatively). We, therefore, audited our hospital practice with the objective of making recommendations for microvascular breast reconstruction patients. PATIENTS AND METHODS: A review was performed of all free flap breast reconstructions between 2014 and 2019. Patients were classified according to hormone medication status at operation. Timings of drug cessation and recommencement were recorded. Thrombotic events, namely flap microvascular thrombosis, deep vein thrombosis, superficial vein thrombosis and pulmonary embolism, were compared. RESULTS: A total of 240 patients had 275 free flaps over five years with 36 receiving hormone therapy within one month prior to surgery, which was discontinued 8.5 days (range: 0-28 days) before surgery. Intraoperative microvascular thromboses (HT 2.0%, NHT 0%, and p = 0.869) and post-operative microvascular complications/flap re-explorations (HT 6.6%, NHT 0%, and p = 0.234) were comparable between the two groups. Systemic venous thromboembolic events were also similar (HT 8.3%, NHT 6.1%, and p = 0.893). Age, BMI, smoking status and preoperative chemotherapy did not influence the incidence of thrombotic complications. CONCLUSION: Hormone therapy did not significantly increase the risk of thromboembolic events. Despite the widespread practice of withholding it for 2 weeks prior to reconstructive surgery, this study does not support such practice being beneficial in terms of thromboembolic events and flap viability. Large-scale trials are needed to establish definitive protocols.

An unusual "venous circle" of the internal mammary vein encountered during microvascular anastomosis and implications for practice.

The internal mammary vessels are commonly used for anastomosis in breast reconstruction. The anatomy when using the 2nd ICS has been shown to be predictable and hence preferentially used by the senior author. We present an unusual case of internal mammary vein bifurcation and immediate confluence forming a 'venous circle'.

The ideal intercostal space for internal mammary vessel exposure during total rib-sparing microvascular breast reconstruction: A critical evaluation.

BACKGROUND: Total rib-preserving free flap breast reconstruction (RP-FFBR) using internal mammary vessel (IMV) recipients usually involves vessel exposure in the second or third intercostal spaces (ICS). Although the third one is more commonly used, no direct comparisons between the two have hitherto been performed. OBJECTIVES: To compare the in-vivo topography and vascular anatomy of second and third ICSs in patients undergoing FFBR using the rib-preservation technique of IMV exposure. METHODS: An analysis of prospectively collected data on intercostal space distance (ISD), number and arrangement of IMVs, location of venous confluence, and vessel exposure time was conducted on a single surgeon's consecutive RP-FFBRs. RESULTS: A total of 296 RP-FFBRs were performed in 246 consecutive patients. The second, third, or both second and third spaces were utilized in 282, 28, and 22 cases, respectively. The ISDs were 20.6 mm ±â€¯3.52 for the second ICS and 14.0 mm ± 4.35 for the third ICS (p<0.0001, CI = 5.17-7.97, t-test). The second versus third ICS vein content was as follows: single 81.4% vs. 74%, dual 18.6% vs. 26%, and confluence 3.7% vs. 13%. The second ICS single vein was medial to the artery in 92.6%. The third ICS single vein was medial to the artery in 88.2% Vessel exposure times for second (47.2 mins ±â€¯26.7) and third (46.5 mins ±â€¯31.4) spaces were similar (p = 0.93). The overall intraoperative anastomotic revision rate was 9.1%, and the postoperative flap re-exploration rate was 4.0%, with 99.7% overall flap success. DISCUSSION AND CONCLUSION: Preferential use of the second ICS is supported by its more predictable vascular anatomy, a broader space for performing the microanastomoses and a higher frequency of a single postconfluence (and thus larger) vein facilitating the microsurgery.

4-Hexyl-1,3-phenylenediol, a nuclear factor-κB inhibitor, improves photodamaged skin and clinical signs of ageing in a double-blinded, randomized controlled trial.

BACKGROUND: The nuclear factor-κB (NF-κB) pathway is a key mediator of inflammation; however, few studies have examined the direct effects of NF-κB inhibition on the skin. OBJECTIVES: To investigate NF-κB activity in cultured human fibroblasts and to investigate the effects of 4-hexyl-1,3-phenylenediol (an NF-κB inhibitor) on elastin and collagen gene expression in vitro and on the clinical appearance of photodamaged skin. METHODS: The amount and activity of NF-κB in human fibroblasts obtained from donors (17-78 years old) was measured after transfection with a NF-κB reporter and a luciferase promoter system. The expression of extracellular matrix (ECM) genes was determined using quantitative polymerase chain reaction. Women with moderate skin photodamage were randomized to daily treatment with a topical lotion containing 4-hexyl-1,3-phenylenediol (n = 30) or vehicle (n = 29) for 8 weeks, with clinical assessments at baseline and weeks 2, 4 and 8. RESULTS: Fibroblasts obtained from donors older than 50 years had higher NF-κB activity compared with cells from younger donors; inhibition of the NF-κB pathway with 4-hexyl-1,3-phenylenediol enhanced the expression of ECM genes. In women, treatment for 8 weeks with 4-hexyl-1,3-phenylenediol significantly improved crow's feet fine lines, cheek wrinkles, age spots, mottled pigmentation and radiance compared with both the vehicle and baseline. Furthermore, treatment with 4-hexyl-1,3-phenylenediol resulted in a twofold greater clinical improvement in overall photodamage compared with the vehicle group. CONCLUSIONS: Inhibition of the proinflammatory NF-κB pathway resulted in increased expression of ECM proteins in vitro and significant clinical improvement in photodamaged skin.

The NeuroD1/BETA2 sequences essential for insulin gene transcription colocalize with those necessary for neurogenesis and p300/CREB binding protein binding.

NeuroD1/BETA2 is a key regulator of pancreatic islet morphogenesis and insulin hormone gene transcription in islet beta cells. This factor also appears to be involved in neurogenic differentiation, because NeuroD1/BETA2 is able to induce premature differentiation of neuronal precursors and convert ectoderm into fully differentiated neurons upon ectopic expression in Xenopus embryos. We have identified amino acid sequences in mammalian and Xenopus NeuroD1/BETA2 that are necessary for insulin gene expression and ectopic neurogenesis. Our results indicate that evolutionarily conserved sequences spanning the basic helix-loop-helix (amino acids [aa] 100 to 155) and C-terminal (aa 156 to 355) regions are important for both of these processes. The transactivation domains (AD1, aa 189 to 299; AD2, aa 300 to 355) were within the carboxy-terminal region, as analyzed by using GAL4:NeuroD1/BETA2 chimeras. Selective activation of mammalian insulin gene enhancer-driven expression and ectopic neurogenesis in Xenopus embryos was regulated by two independent and separable domains of NeuroD1/BETA2, located between aa 156 to 251 and aa 252 to 355. GAL4:NeuroD1/BETA2 constructs spanning these sequences demonstrated that only aa 252 to 355 contained activation domain function, although both aa 156 to 251 and 300 to 355 were found to interact with the p300/CREB binding protein (CBP) coactivator. These results implicate p300/CBP in NeuroD1/BETA2 function and further suggest that comparable mechanisms are utilized to direct target gene transcription during differentiation and in adult islet beta cells.

Regulation of insulin-like growth factor I biosynthesis in porcine granulosa cells.

In the present studies we examined the regulation of insulin-like growth factor I (IGF-I) expression in porcine granulosa cells in vitro. Using Northern analysis and ribonuclease protection assays with exon-specific probes, we identified the IGF-I messenger RNA (mRNA) transcripts present in these cells under basal and hormone-stimulated conditions. We also assessed changes in secreted IGF-I using Western blots and correlated the change in protein secretion after hormone treatment with changes in mRNA levels. By analogy to the human IGF-I gene and its transcription, two major transcripts of approximately 1 and 7.5 kilobases, seen in freshly isolated granulosa cells and follicle wall and in single passaged granulosa (MDGp1) cells, most likely correspond to IGF-IA. Minor transcripts of 3-4 kilobases, which appeared after FSH or forskolin treatments or in control cells after long exposure of the autoradiographs, were attributed to incompletely processed RNA precursors. Ribonuclease protection assay analysis using probes to detect alternative use of exon 5 or exon 6 indicated that most, if not all, of the transcripts contained only exon 6 sequence (IGF-IA). Both class 1 and class 2 transcripts were identified using exon 1- and exon 2-specific probes, respectively. GH increased steady state levels of IGF-I mRNA 3-fold, FSH increased it approximately 10-fold, and forskolin maximally increased it 12- to 15-fold. Estradiol had no effect alone or in combination with the other treatments. All treatments that increased IGF-I mRNA coordinately increased both class 1 and class 2 transcripts, with the increase in class 1 greater than that in class 2. Multiple forms of IGF-I protein were seen under basal conditions and after hormone treatment. These were identified based on mRNA analysis and biochemical methods as both glycosylated and nonglycosylated IGF-IA prohormone, incompletely processed forms of prohormone, and the mature peptide. Changes in the levels of total protein were similar to the changes in mRNA (GH, 3-fold; FSH and forskolin, 10- to 20-fold). All forms of the protein changed coordinately, suggesting that these hormones had no major effect on the intracellular processing mechanism. IGF-binding protein-3 was able to bind to all IGF-I forms. These data conclusively demonstrate FSH and GH induction of ovarian IGF-I. The porcine granulosa cell culture system used in these studies should be an excellent system for studying the hormonal regulation of IGF-I expression.

Expression of the messenger ribonucleic acids for insulin-like growth factor-I and insulin-like growth factor binding proteins in porcine corpora lutea.

In the pig, the corpus luteum (CL) can develop and function autonomous of pituitary gonadotropins for approximately 12 days. We hypothesized that the insulin-like growth factor (IGF) system may play an autocrine/paracrine luteotrophic role(s) during this period. In this study, we monitored the expression (i.e., steady-state levels of mRNAs) of IGF-I and IGF binding proteins (IGFBP)-2, -3, -4, -5, and -6 mRNAs in whole CL and in small and large luteal cells on Days 4-16 of the estrous cycle. CL were dissociated with collagenase, and large and small luteal cells were isolated by centrifugal elutriation. Whole CL and luteal cells were extracted to isolate total or poly(A)+ RNA, which was subjected to Northern and/or dot-blot analyses using [32P]-labeled cDNA probes for IGF-I and IGFBP-2, -3, -4, -5, and -6. Northern blots showed readily detectable transcripts for IGF-I (6.7 and 0.9 kb), IGFBP-2 (1.8 kb), IGFBP-3 (2.8 kb), IGFBP-4 (2.6 kb), and IGFBP-5 (6.0 kb), but not for IGFBP-6. IGFBP-3 and -5 transcripts were observed mainly in small luteal cells, while IGFBP-2 and -4 were seen in both cell types. Dot-blot analyses for IGF-I and IGFBP-3 mRNAs were performed on total RNA from small and large luteal cells; blots were counter-probed with 3-phosphoglyceraldehyde dehydrogenase (p-GAD) cDNA to assess RNA quantity and quality. IGF-I mRNA (ratio IGF-I:p-GAD mRNA) expression was approximately 2-fold greater in small than in large luteal cells on Days 4-10. However, steady-state levels of IGF-I mRNA in small, but not large, luteal cells decreased significantly on Days 12-16 (vs. Days 4-10). IGFBP-3 mRNA expression was significantly greater (approximately 3-fold) in small than in large luteal cells but did not vary significantly between Days 4-10 and 12-16 for either cell type. We conclude that porcine CL express mRNAs for IGF-I and IGFBP-2, -3, -4, and -5, and that while small luteal cells are the major sources of IGF-I and IGFBP-3 and -5, IGFBP-2 and -4 appear to be expressed to approximately the same extent in small and large luteal cells. These results further suggest that the IGF-I/IGF system may have autocrine/paracrine regulatory actions in CL development/function in the pig.

Insulin-like growth factor binding protein-3 inhibits porcine granulosa cell function in vitro.

Recombinant human insulin-like growth factor binding protein-3 (rhIGFBP-3) effects on basal, insulin-like growth factor I (IGF-I)-, and follicle-stimulating hormone (FSH)-stimulated progesterone (P4) secretion and [3H]aminoisobutyric acid (AIB) uptake by primary porcine granulosa cells (MDGs) and MDGs that have been passaged once (MDGp1) were assessed. Cells were treated concurrently or were preincubated with rhIGFBP-3 followed by treatment. rhIGFBP-3 had no effect on MDG or MDGp1 cell numbers after 24 h. Cotreatment with rhIGFBP-3 inhibited P4 secretion after treatment with FSH, IGF-I, and FSH plus IGF-I. FSH did not stimulate [3H]AIB uptake. However, the IGF-I-stimulated increase in [3H]AIB uptake was completely prevented by concurrent treatment with IGFBP-3. Preincubation of MDGp1 cells with IGFBP-3 dose dependently inhibited FSH- and IGF-I-stimulated P4 secretion. This inhibition was associated with increased cell association of the binding protein and increased IGF-I binding to the cells. These results indicate that IGFBP-3 is inhibitory to a variety of crucial functions in porcine granulosa cells, supporting a role for it in the regulation of granulosa cell function.

Changes in the messenger ribonucleic acid for insulin-like growth factor-I and -II in the porcine testis during and between two waves of testicular development.

The boar testis was used as a model for examining the possible role of production of the insulin-like growth factors (IGF) in steroidogenesis and/or testicular growth as testicular development occurs in waves. Blood and testes were sampled from boars at different ages (100-102 days of gestation; 7, 19, and 30 days; and 10 and 25 wk), selected to occur during and between the last two waves of testicular development. Serum was analyzed for testosterone and RNA was extracted from the testes for Northern and dot-blot analysis of IGF mRNA. Testosterone concentrations declined (p = 0.01) from 7 days to 10 wk of age and rebounded at 25 wk, indicating completion of the second and third waves of testicular development. The quantity of testicular mRNA for IGF-I increased gradually with age, and that for IGF-II decreased. We therefore conclude that regulation of the expression of the mRNAs for IGF-I and -II in the pig testis is not a function of either the waves of testicular development or the level of steroidogenesis.

Effects of growth hormone and gonadotropin on the insulin-like growth factor system in the porcine ovary.

The effects of growth hormone (GH) +/- pregnant mare's serum gonadotropin (PMSG) on levels of insulin-like growth factor (IGF)-I and -II and IGF binding protein (BP)-2 and -3 in serum and follicular fluid (FFI) and on the expression of their mRNA in the ovaries of prepubertal gilts were determined. Steroids in FFI were also quantified. In the first experiment, GH, given for either 20 or 40 days, caused a distinct (threefold, p < 0.05) increase in IGF-I in both serum and FFI with no change in the FFI:serum ratio (0.65). Effects of GH on IGF-II were opposite, with a drop in circulating and FFI levels (p < 0.05). In contrast to data for IGF-I, FFI levels were higher than those in serum for IGF-II (1.42, FFI:serum); IGF-II levels and the ratio fell after GH treatment. GH for either 20 days or 40 days increased serum IGBP-3 to 140% and 250% of control values while decreasing serum IGFBP-2 by 46% and 31%, respectively (p < 0.001). FFI IGFBP-3 was increased to a similar extent by GH (p < 0.005), but IGFBP-2 was not affected. Neither progesterone (P4) nor estradiol (E2) was affected by treatment with GH. However, androstenedione (A4) was decreased by 20-day and 40-day GH treatment relative to the respective controls (p < 0.05). In the second experiment, PMSG resulted in a modest (28%) increase in intrafollicular IGF-I (p < 0.06).(ABSTRACT TRUNCATED AT 250 WORDS)

Expression of the mRNAs for the insulin-like growth factors and their binding proteins during development of porcine ovarian follicles.

We examined the expression of the mRNAs for the insulin-like growth factors (IGFs) and two of their binding proteins (BPs), IGFBP-2 and IGFBP-3, in individual follicles of the porcine ovary. Follicular development was synchronized with a progestin (altrenogest). Individual follicles were isolated on days 1, 3, 5 and 7 after progestin withdrawal. No IGFBP-3 mRNA was detected. While IGF-II mRNA was easily detected, the levels of expression did not change. IGF-I and IGFBP-2 mRNAs increased and decreased, respectively, with follicle development until day 7 when IGF-I expression declined. Regression analysis of IGF-I and IGFBP-2 mRNA expression was performed to assess the relative strength of correlations with day, diameter and steroid concentrations as covariates. IGFBP-2 mRNA was correlated with both day and diameter (r = -.713 and -.705, respectively, n = 24) and neither estrogen (E2) nor progesterone (P4) contributed to the fit. While IGF-I mRNA expression was correlated to both day (r = .483) and diameter (r = .587), the strongest predictor was E2 concentration (r = .694, n = 27). In conclusion, the expression of IGF-I and IGFBP-2 mRNAs in the ovarian follicle are discordantly regulated during follicular growth and maturation. The observed changes in these parameters should result in increased bioavailable IGF-I. This supports a pivotal autocrine/paracrine role for these factors during follicle growth and development.

Divergent actions of prostaglandins-E2 and -F2 alpha on the regulation of insulin-like growth factor-binding protein-3 in luteinized granulosa cells.

Insulin-like growth factor (IGF)-I synergizes with gonadotropin to further stimulate ovarian steroidogenesis. In contrast, the IGF-binding proteins (IGFBPs) produced by granulosa cells have been shown to antagonize the stimulatory actions of the IGFs and gonadotropins. The purpose of the present study was to examine the effects on IGFBP-3 production of prostaglandin (PG)-E2, a compound known to stimulate luteal function and prevent/delay luteal regression (a luteotropic compound), and PGF2 alpha, a compound known to be luteolytic. PGF2 alpha significantly stimulated IGFBP-3 production to 2.6-fold of control (P < 0.05) while PGE2 attenuated its production to half of control (P < 0.05). In contrast to the effects of IGFBP-3, PGE2 stimulated progesterone production to 8-fold of control (P < 0.05) while PGF2 alpha had no effect. Possible mechanisms of action of PGE2 and PGF2 alpha were also examined. PGE2, but not PGF2 alpha, stimulated cAMP accumulation which has been previously shown to inhibit IGFBP-3 production. PGF2 alpha is suspected to act via activation of protein kinase-C. However, a phorbol ester did not mimic PGF2 alpha's action toward IGFBP-3. This study demonstrated that PGE2 and PGF2 alpha conversely modulate IGFBP-3 production. Since IGFBPs have been shown to antagonize gonadotropin and IGF actions, this action of the prostaglandins may impact on the synergism between IGFs and gonadotropin necessary for follicular growth and luteal function.

The role of insulin-like growth factors and epidermal growth factor-related peptides in intraovarian regulation in the pig ovary.

The autocrine and paracrine role of the insulin-like growth factors (IGFs) and epidermal growth factor (EGF)-related peptides in pig ovary are reviewed. For convenience, each of these regulatory systems is divided into several interactive components: regulated expression of the growth factors, growth factor reception at the cell surface and intracellular action of the growth factors. In addition, the concept of regulated bioavailability and targeting of growth factors in the extracellular space is developed as an important control locus and area for future study. With regard to the IGF system, these components include two ligands--IGF-I and -II, both expressed in the porcine ovary--and the possibility of three receptors. IGF-I and the type I IGF receptor appear to be the most important in stimulating ovarian function and amplifying hormone action. In addition, the 'set-point' of the ovarian IGF system may be determined by the activity of several IGF-binding proteins (IGFBPs). At least four of these proteins are expressed in the pig ovary. Studies of their regulation and action in ovarian cells indicate that they can function as antagonists to FSH and the IGFs. However, preliminary evidence suggests a more dynamic model in which these proteins may direct the site and timing of IGF effects. There are fewer data on the EGF system. At least four EGF-related peptides are expressed in pig ovaries, but insufficient information is available to predict their physiological regulation. These peptides are potent mitogens for ovarian cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Expression of insulin-like growth factor-binding protein-2 and -3 messenger ribonucleic acid in the porcine ovary: localization and physiological changes.

Insulin-like growth factor-binding protein (IGFBP)-2 and -3 are the most prevalent IGFBPs in porcine follicular fluid, as determined on ligand blots, but little is known about the localization and regulation of their synthesis in vivo. This study was designed to investigate the localization and cyclic regulation of the mRNA for these two IGFBPs in the porcine ovary, RNA was extracted from whole ovaries morphologically classified as immature, preovulatory, and luteal. Northern hybridization analysis of this RNA showed no significant difference in the expression of IGFBP-2 mRNA in these ovaries (OD for preovulatory, luteal, and immature ovaries, 0.076 +/- 0.01, 0.071 +/- 0.01, and 0.10 +/- 0.008/micrograms RNA, respectively). IGFBP-3 mRNA was not different in immature and preovulatory ovaries, but was 10-fold greater (P less than 0.025) in luteal ovaries. Northern analysis of RNA extracted from ovaries also showed no significant change in IGFBP-2 mRNA on days (d) 11, 16, and 21 of the estrous cycle. IGFBP-3 mRNA tended to decrease between d11-16 with the onset of luteal regression and was significantly decreased in d21 preovulatory ovaries to 22% of the values in d11 ovaries. Granulosa, thecal, and luteal cells were also analyzed for IGFBP mRNA. IGFBP-2 mRNA was most abundant in granulosa cells, lower in thecal cells, and lowest in luteal cells. No IGFBP-3 mRNA could be detected in granulosa cells, and luteal cells expressed 15- to 63-fold greater levels than thecal cells. These results show that IGFBP-2 and -3 mRNAs are expressed in specific ovarian cell types and that their expression appears to be independently regulated during the reproductive cycle. This provides further evidence for the importance of these proteins as paracrine/autocrine regulators of ovarian function.

Gonadotropin and cAMP modulation of IGE binding protein production in ovarian granulosa cells.

Porcine granulosa cells (GC) produce insulin-like growth factor (IGF) binding protein (BP)-3 and IGFBP-2 in culture. A gonadotropin, follicle-stimulating hormone (FSH), dramatically inhibited GC production of these IGFBPs in control cultures and in cultures stimulated by insulin plus epidermal growth factor (EGF) or IGF-I plus EGF. Stimulators of adenylate cyclase (forskolin, cholera toxin) and a derivative of adenosine 3',5'-cyclic monophosphate (cAMP), 8-bromoadenosine 3',5'-cyclic monophosphate, inhibited IGFBP synthesis in a manner similar to FSH. In contrast, the antagonist of cAMP action, (R)-p-adenosine 3',5'-cyclic phosphorothioate [(R)-p-cAMPS], significantly stimulated production of IGFBP-3 and IGFBP-2 compared with controls. This stimulatory effect of (R)-p-cAMPS was counteracted by cotreatment with FSH in a dose-dependent manner. Finally, treatment of GC cultures with FSH plus 3-isobutyl-1-methylxanthine resulted in a significant reduction in cellular content of mRNA coding for IGFBP-3 with no change in IGFBP-2 mRNA. In summary, agents that elevate intracellular cAMP were found to mimic the effects of FSH on IGFBP production.

The ovarian insulin-like growth factors, a local amplification mechanism for steroidogenesis and hormone action.

The importance of the ovarian insulin-like growth factors (IGFs) has been suggested by data from numerous laboratories and several approaches in the last several years. In the aggregate, these data indicate that this system could function as an important local amplification mechanism for steroidogenesis and gonadotropin action. Studies supporting this hypothesis have described several interacting components of this autocrine/paracrine system. First, the several types of ovarian cells possess an IGF-response system, which includes receptors for IGFs and an effective intracellular transduction system. The IGFs can promote growth and/or differentiation of ovarian cells, and their predominant actions depend on the nature of the cells and the presence of additional modulating factors. The biochemical events leading to enhanced steroidogenesis are now understood in considerable detail and include induction of several steps in the cAMP-dependent steroidogenic cascade. The second component of the ovarian IGF system comprises hormone-responsive local production of IGFs. Both IGF-I and IGF-II may be secreted; gonadotropins, gonadal steroids and locally produced growth factors can regulate the IGF system at this level. Finally, ovarian cells secrete a heterogeneous and complex family of IGF-binding proteins (IGFBPs). These proteins can impact on multiple ovarian functions in a manner which is generally opposite to that of the IGFs themselves. As is the case for the IGFs, the secretion of these proteins by ovarian cells is regulated by gonadotropins and locally produced ovarian factors. Collectively, these several components provide an integrated, synergistically cooperative local network to promote gonadotropin-dependent growth and differentiation in the ovary.

The effect of severing the spinal cord of fetal lambs on length of gestation.

Lambs with spinal cords that had been severed near the head at 50 days of gestation were born after a shortened gestation of about 128 days. Lambs with severed spinal cords born as a twin with an intact lamb had a normal gestation of 148 days. The presence of an intact spinal cord or the signals that it might carry apparently influence the length of gestation in the ewe.