Determinants of optimal mastectomy skin flap thickness.

BACKGROUND: There is a limited evidence base to guide surgeons on the ideal thickness of skin flaps during mastectomy. Here the literature relevant to optimizing mastectomy skin flap thickness is reviewed, including anatomical studies, oncological considerations, factors affecting viability, and the impact of surgical technique and adjuvant therapies. METHODS: A MEDLINE search was performed using the search terms 'mastectomy' and 'skin flap' or 'flap thickness'. Titles and abstracts from peer-reviewed publications were screened for relevance. RESULTS: A subcutaneous layer of variable thickness that contains minimal breast epithelium lies between the dermis and breast tissue. The thickness of this layer may vary within and between breasts, and does not appear to be associated with obesity or age. The existence of a distinct layer of superficial fascia in the breast remains controversial and may be present in only up to 56 per cent of patients. When present, it may not be visible macroscopically, and can contain islands of breast tissue. As skin flap necrosis occurs in approximately 5 per cent of patients, a balance must be sought between removing all breast tissue at mastectomy and leaving reliably viable skin flaps. CONCLUSION: The variable and unpredictable thickness of the breast subcutaneous layer means that a single specific universal thickness for mastectomy skin flaps cannot be recommended. It may be that the plane between the subdermal fat and breast parenchyma is a reasonable guide for mastectomy flap thickness, but this may not always correspond to a subcutaneous fascial layer.

Seminal fluid and immune adaptation for pregnancy--comparative biology in mammalian species.

Seminal fluid delivered to the female reproductive tract at coitus not only promotes the survival and fertilizing capacity of spermatozoa, but also contains potent signalling agents that influence female reproductive physiology to improve the chances of conception and reproductive success. Male to female seminal fluid signalling occurs in rodents, domestic and livestock animals, and all other mammals examined to date. Seminal plasma is instrumental in eliciting the female response, by provision of cytokines and prostaglandins synthesized in the male accessory glands. These agents bind to receptors on target cells in the cervix and uterus, activating changes in gene expression leading to functional adaptations in the female tissues. Sperm also interact with female tract cells, although the molecular basis of this interaction is not yet defined. The consequences are increased sperm survival and fertilization rates, conditioning of the female immune response to tolerate semen and the conceptus, and molecular and cellular changes in the endometrium that facilitate embryo development and implantation. Studies in porcine, equine, bovine, ovine and canine species all show evidence of male-female signalling function for seminal fluid. There are variations between species that relate to their different reproductive strategies and behaviours, particularly the site of seminal fluid deposition and female reproductive tract anatomy. Although the details of the molecular mechanisms require more study, the available data are consistent with both the sperm and plasma fractions of seminal fluid acting in a synergistic fashion to activate inflammation-like responses and downstream female tract changes in each of these species. Insight into the biological function and molecular basis of seminal fluid signalling in the female will inform new interventions and management practices to support optimal reproductive outcomes in domestic, livestock and endangered animal species.

Two approaches to discovering and developing new drugs for Chagas disease.

This review will focus on two general approaches carried out at the Sandler Center, University of California, San Francisco, to address the challenge of developing new drugs for the treatment of Chagas disease. The first approach is target-based drug discovery, and two specific targets, cytochrome P450 CYP51 and cruzain (aka cruzipain), are discussed. A 'proof of concept' molecule, the vinyl sulfone inhibitor K777, is now a clinical candidate. The preclinical assessment compliance for filing as an Investigational New Drug with the United States Food and Drug Administration (FDA) is presented, and an outline of potential clinical trials is given. The second approach to identifying new drug leads is parasite phenotypic screens in culture. The development of an assay allowing high throughput screening of Trypanosoma cruzi amastigotes in skeletal muscle cells is presented. This screen has the advantage of not requiring specific strains of parasites, so it could be used with field isolates, drug resistant strains or laboratory strains. It is optimized for robotic liquid handling and has been validated through a screen of a library of FDA-approved drugs identifying 65 hits.

NS1 DNA vaccination protects against Zika infection through T cell-mediated immunity in immunocompetent mice.

The causal association of Zika virus (ZIKV) with microcephaly, congenital malformations in infants, and Guillain-Barré syndrome in adults highlights the need for effective vaccines. Thus far, efforts to develop ZIKV vaccines have focused on the viral envelope. ZIKV NS1 as a vaccine immunogen has not been fully explored, although it can circumvent the risk of antibody-dependent enhancement of ZIKV infection, associated with envelope antibodies. Here, we describe a novel DNA vaccine encoding a secreted ZIKV NS1, that confers rapid protection from systemic ZIKV infection in immunocompetent mice. We identify novel NS1 T cell epitopes in vivo and show that functional NS1-specific T cell responses are critical for protection against ZIKV infection. We demonstrate that vaccine-induced anti-NS1 antibodies fail to confer protection in the absence of a functional T cell response. This highlights the importance of using NS1 as a target for T cell-based ZIKV vaccines.

Cytokines in rodent reproduction and the cytokine-endocrine interaction.

Insights derived from recent studies employing rodent models demonstrate that the synthesis of pluripotent cytokines is an important function of resident cells in the female reproductive tract. Through steroid hormone regulated secretion of these mediators, resident cells appear to coordinate the recruitment and action of leukocytes that are centrally implicated in the dramatic remodelling processes characteristic of reproductive events.

Use of a transdermal matrix patch of buprenorphine in cats: preliminary pharmacokinetic and pharmacodynamic data.

Six domestic shorthair cats, aged three to four years and weighing 5.1 to 7.4 kg, were used to assess the thermal antinociceptive effect of a transdermal buprenorphine patch, designed to supply 35 mug buprenorphine/hour, which was applied to the shaved thorax. The cats' thermal thresholds were tested before the patch was applied and two, four, six, eight, 10, 12, 14 and 16 hours after it had been applied, and then every six hours until it was removed after 72 hours, and for a further 24 hours afterwards. Blood was collected at each time to measure the plasma concentration of buprenorphine. The patches did not produce a significant change in the thermal thresholds of the cats throughout the testing period. The mean (sd) peak plasma buprenorphine concentration was 10 (0.81) ng/ml.

Protection against maternal infection-associated fetal growth restriction: proof-of-concept with a microbial-derived immunomodulator.

Infection-associated inflammatory stress during pregnancy is the most common cause of fetal growth restriction and/or miscarriage. Treatment strategies for protection of at-risk mothers are limited to a narrow range of vaccines, which do not cover the bulk of the common pathogens most frequently encountered. Using mouse models, we demonstrate that oral treatment during pregnancy with a microbial-derived immunomodulator (OM85), currently used clinically for attenuation of infection-associated airway inflammatory symptoms in infants-adults, markedly reduces risk for fetal loss/growth restriction resulting from maternal challenge with bacterial lipopolysaccharide or influenza. Focusing on LPS exposure, we demonstrate that the key molecular indices of maternal inflammatory stress, notably high levels of RANTES, MIP-1α, CCL2, KC, and G-CSF (granulocyte colony-stimulating factor) in gestational tissues/serum, are abrogated by OM85 pretreatment. Systems-level analyses conducted in parallel using RNASeq revealed that OM85 pretreatment selectively tunes LPS-induced activation in maternal gestational tissues for attenuated expression of TNF, IL1, and IFNG-driven proinflammatory networks, without constraining Type1-IFN-associated networks central to first-line antimicrobial defense. This study suggests that broad-spectrum protection-of-pregnancy against infection-associated inflammatory stress, without compromising capacity for efficient pathogen eradication, represents an achievable therapeutic goal.

Resource implications of risk-reducing mastectomy and reconstruction.

AIM: Risk-reducing mastectomy (RRM) is on the increase, now frequently combined with breast reconstruction (BR). However, the resource implications associated with bilateral mastectomy and reconstruction are unknown. This study assessed the overall cost of performing risk-reducing surgery. METHODS: All cases of RRM and BR performed between 1991 and 2011 at this hospital were identified from a prospectively collected database. All patients undergoing bilateral mastectomy were included, when at least one mastectomy was risk-reducing. Overall treatment costs for all surgical procedures, complications, revisional procedures and outpatient attendances were calculated and compared to the National Tariff allowed. Mann-Whitney U and Fischer's exact tests were used to calculate levels of significance. RESULTS: Fifty patients underwent bilateral mastectomy and BR (median follow up 20 [range 1-106] months), 72 were Latissimus Dorsi reconstructions (LDR) and 28 were Subpectoral reconstructions (SPR). LDR took longer than SPR (p = 0.001), with a greater length of stay (p = 0.024). Nine percent of patients returned to theatre for early complications, but the type of BR did not influence the early complication rate (LDR versus SPR, p = 0.345) or the need for additional unplanned procedures (LDR versus SPR, p = 0.671). The overall mean cost for bilateral RRM and BR was £14,797 per patient. The inpatient cost for bilateral RRM and LDR was £10,082 compared with £5,905 SPR. Both procedures exceeded the £5,697 tariff allowed in the UK. CONCLUSION: Bilateral RRM and BR is a safe procedure, but the resource implications are considerable and exceed the tariff allowed, particularly when performing more complex techniques.

A review of fundamental principles for animal models of DOHaD research: an Australian perspective.

Epidemiology formed the basis of 'the Barker hypothesis', the concept of 'developmental programming' and today's discipline of the Developmental Origins of Health and Disease (DOHaD). Animal experimentation provided proof of the underlying concepts, and continues to generate knowledge of underlying mechanisms. Interventions in humans, based on DOHaD principles, will be informed by experiments in animals. As knowledge in this discipline has accumulated, from studies of humans and other animals, the complexity of interactions between genome, environment and epigenetics, has been revealed. The vast nature of programming stimuli and breadth of effects is becoming known. As a result of our accumulating knowledge we now appreciate the impact of many variables that contribute to programmed outcomes. To guide further animal research in this field, the Australia and New Zealand DOHaD society (ANZ DOHaD) Animals Models of DOHaD Research Working Group convened at the 2nd Annual ANZ DOHaD Congress in Melbourne, Australia in April 2015. This review summarizes the contributions of animal research to the understanding of DOHaD, and makes recommendations for the design and conduct of animal experiments to maximize relevance, reproducibility and translation of knowledge into improving health and well-being.

The role of cytokines in gestation.

Lymphohemopoietic cytokines are now recognized to be central participants in the cellular communication events underlying the complex and dynamic remodeling processes required to accommodate the semiallogeneic conceptus during mammalian reproduction. Cytokines are identified to be particular importance in mediating communications between the conceptus and maternal cells, particularly the uterine epithelium and infiltrating leukocytes, both prior to implantation and as the placenta develops. In this review we summarize recent experimental data concerning the synthesis of various cytokines in uterine and conceptus-derived tissues and highlight current hypotheses for their roles in establishing and maintaining successful pregnancy. It is concluded that complex and finely balanced cytokine networks underpin precise regulatory mechanisms controlling the rate and degree of conceptus development and invasion into maternal tissues.

Effect of systemic lidocaine on visceral and somatic nociception in conscious horses.

REASONS FOR PERFORMING STUDY: Commonly used analgesics (nonsteroidal anti-inflammatory agents, opioids and alpha2-agonists) have unwanted side effects. An effective alternative with minimal adverse effects would benefit clinical equine pain management. OBJECTIVES: To compare the effect of lidocaine or saline on duodenal and rectal distension threshold pressure and somatic thermal threshold in conscious mature horses. HYPOTHESIS: Systemically administered lidocaine would increase somatic and visceral nociceptive thresholds. METHODS: Lidocaine (2 mg/kg bwt bolus followed by 50 microg/kg bwt/min for 2 h) or saline was administered to 6 horses each carrying a permanently implanted gastric cannula, in a randomised, blinded cross-over design. Thermal threshold was measured using a probe containing a heater element placed over the withers which supplied heat until the horse responded. A barostatically controlled intraduodenal balloon was distended until a discomfort response was obtained. A rectal balloon was inflated until extruded or signs of discomfort noted. RESULTS: Thermal threshold was increased significantly 30 and 90 mins after the start of lidocaine infusion. There was no change in duodenal distension pressure and a small but clinically insignificant change in colorectal distension pressure in the lidocaine group. CONCLUSIONS: At the dose used, systemically administered lidocaine produced thermal antinociception but minimal changes in visceral nociception. POTENTIAL RELEVANCE: At these doses, lidocaine may play a role in somatic analgesia in horses.

Retarded development of exoerythrocytic stages of the rodent malaria parasite Plasmodium berghei in human hepatoma cells by extracts from Dioncophyllaceae and Ancistrocladaceae species.

Retarded development of exoerythrocytic stages of the rodent malaria parasite Plasmodium berghei in human hepatoma cells by extracts from Dioncophyllaceae and Ancistrocladaceae species. International Journal for Parasitology 27: 29-32. Naphthylisoquinoline alkaloid-containing extracts (10 micrograms ml-1) of species belonging to the Dioncophyllaceae and the Ancistrocladaceae, 2 small tropical plant families, display pronounced in vitro activities against exoerythrocytic stages of Plasmodium berghei (Anka), developing in human hepatoma cells (Hep G2). The highest activities were obtained with CH2Cl2 root and bark extracts, and a CH2Cl2/NH3 leaf extract from Triphyophyllum peltatum, a CH2Cl2/NH3 root extract from Ancistrocladus abbreviatus, and a CH2Cl2 leaf extract from A. tectorius. The degrees of growth inhibition ranged within 27.7-70.0%. The commercially available drug primaquine diphosphate (25 micrograms ml-1) caused a comparable effect (62.1%) in the same test system.

Altered placental development in interleukin-10 null mutant mice.

Placental morphogenesis and nutrient transfer function are regulated by growth factors at the foeto-maternal interface. Interleukin (IL)-10, expressed in the decidua and placenta, is implicated in regulating extravillous cytotrophoblast invasion through inhibiting matrix metalloproteinase expression and influencing the quality of the maternal immune response. Our laboratory has previously found that IL-10 deficiency in both the mother and foetus increases foetal weight at day 18 without altering placental weight suggesting that the placenta has a greater functional capacity when IL-10 is absent. The present study has used IL-10 null mutant (IL-10-/-) mice to investigate the role of IL-10 in placental development. Placental structure was assessed in adult virgin IL-10-/- or wild-type (IL-10+/+) mice mated with males of the same genotype and sacrificed at day 18 of gestation. Mid-sagittal cross sections of placental tissue were stained with Masson's trichrome or immuno-labelled with MTS-12 and pan-cytokeratin reactive antibodies to identify foetal endothelial cells and trophoblasts, respectively, and examined with video image analysis. IL-10 deficiency increased the total cross sectional area of the placenta by 28 per cent (IL-10-/- n=22 placentae from 8 dams, IL-10+/+n =21 placentae from 9 dams, P=0.026), principally through increasing the cross sectional area of placental labyrinth by 37 per cent (P=0.025). The proportion of maternal blood space in the labyrinth was increased by 26 per cent (P=0.001) and that of trophoblast was decreased by 16 per cent (P=0.001) in IL-10-/- placentae. The surface area of trophoblast per gram of labyrinth was increased by 41 per cent (P=0.0005) in IL-10-/- placentae. In the absence of IL-10, structural correlates of placental function are enhanced consistent with concomitant increases in foetal growth. These data indicate that IL-10 is a regulator of placental morphogenesis, acting to retard expansion of the placental labyrinth and to modify the architecture of the maternal blood sinuses. Since previous studies have paradoxically shown that postnatal growth is impaired in IL-10 null mutants, these observations are consistent with a role for IL-10 in regulating placental development and foetal programming.

Uterine macrophages and environmental programming for pregnancy success.

Macrophages are ubiquitous cells with an impressive range of functions. These include phagocytosis and coordination of the initiation and effector phases of immune responses, as well as production of bioactive proteins and lipids that profoundly influence cell growth, differentiation and function. Macrophages are highly individualized in tissues, where their activities are a reflection of targeting by systemic and local environmental signals. This review focuses on recent studies where uterine macrophage population densities and distribution have been mapped, chemotaxis, differentiation and activation have been investigated and production of potent effector molecules has been explored. Evidence supporting a major role for female sex steroid hormones and the uterine growth factors they control in governing these features of uterine macrophages is presented.

Reduction of ovulation rate in the rat by administration of a neutrophil-depleting monoclonal antibody.

To investigate the effect of systemic neutrophil depletion on ovulation rate, rats were synchronised with eCG and hCG, and concurrently were administered neutrophil-specific, cytotoxic RP-3 monoclonal antibody (mAb), or an irrelevant, class-matched mAb. Neutrophils in the peripheral blood and in the thecal-luteal area of corpora lutea were detected by immunohistochemical analysis with the neutrophil-specific mAb MCA149 and were found to be reduced in number by 70% and 38% respectively following RP-3 treatment compared to the control group. Ovulation rate, as assessed by counting the number of oocytes in the ampullary region of the oviduct 20 h after hCG administration, was found to be reduced by 27% in the neutrophil-depleted rats. This result provides further evidence that neutrophilic granulocytes play an active role in ovulation in the rat.

The effect of restricted nutrition on uterine macrophage populations in mice.

The abundant macrophage populations present in the endometrium are implicated in the tissue remodelling events and immunological changes necessary for pregnancy. Using two regimens of restricted nutrition (95 and 88% of ad libitum intake for 19 days), we have shown that moderately reduced food consumption can dramatically alter the number of endometrial macrophages and their immunoaccessory function in mice. Restricted nutrition also interfered with the estrous cycle, but the effects on endometrial macrophages were more extensive and qualitatively different than could be explained by diminished ovarian steroid hormone activity. Significantly less F4/80+ and Ia+ cells were found in the endometrium of food restricted mice than in ad libitum mice at the same estrous cycle stage. In the more severely restricted mice the losses were even greater than those seen after ovariectomy. In ad libitum fed animals, uterine but not peritoneal macrophages showed an ovarian hormone-dependent inhibitory phenotype in a splenocyte mitogenesis assay. Macrophages derived from both locations exhibited greater immunostimulatory activity following restricted nutrition. We conclude that endometrial macrophage populations are influenced by nutritional status and this may be mediated through both steroid hormone-dependent and -independent mechanisms. Nutritionally induced aberrations in the number or behaviour of endometrial macrophages during the estrous cycle or in early pregnancy could have important implications for the quality of the pre- and peri-implantation environment and the maternal immune response to pregnancy.

Granulocyte-macrophage colony-stimulating factor (GM-CSF) targets myeloid leukocytes in the uterus during the post-mating inflammatory response in mice.

Factors in seminal plasma elicit a surge of GM-CSF expression in uterine epithelial cells after mating in mice. This study investigates the nature of the endometrial cell populations targeted by epithelial GM-CSF. In quantitative RT-PCR studies, expression of the alpha-subunit of the GM-CSF receptor (GM-CSF-R) parallelled GM-CSF expression, being maximal during the 48 h period after mating and declining thereafter. Expression of mRNA encoding beta-common chain (AIC2B) also increased after mating and remained high until the time of embryo implantation on day 4 of pregnancy. Cells expressing GM-CSF receptors were identified in sections of uterus on the day after mating using 125I-GM-CSF, and were located predominantly in the endometrial stroma subjacent to the luminal epithelium, co-localising with abundant populations of myeloid leukocytes. Cells expressing GM-CSF receptor were identified as macrophages, granulocytes and putative dendritic cells by flow cytometric analysis using lineage and receptor subunit specific antibodies. Recombinant GM-CSF injected into the uterine lumen of ovariectomised mice was found to elicit a dose-dependant accumulation of macrophages and granulocytes in the endometrium, in a pattern of distribution comparable to that seen in uteri after natural mating. Together, these data indicate a role for epithelial cell-derived GM-CSF in mediating the recruitment and potentially in modifying the behaviour of uterine leukocytes during the post-mating inflammatory response in mice.

Tumor necrosis factor alpha in the human ovary: presence in follicular fluid and effects on cell proliferation and prostaglandin production.

OBJECTIVE: To examine the concentration of tumor necrosis factor alpha (TNF alpha) in human follicular fluid (FF) and its effects on cultured human granulosa-lutein cells. DESIGN: The concentration of TNF alpha in FF from hyperstimulated cycles and in conditioned media from cultured granulosa-lutein cells was measured by radioimmunoassay (RIA) and bioassay using L929 cells. The effects of recombinant human TNF alpha (rTNF alpha) on proliferation and production of progesterone (P) and prostaglandin (PG, PGE2, and PGF2 alpha) by cultured human granulosa-lutein cells were assessed. SETTING: In vitro fertilization and embryo transfer (IVF-ET) program at Reproductive Medicine Unit, The Queen Elizabeth Hospital, Woodville, South Australia, Australia. PATIENTS: Twenty-five women undergoing IVF-ET for tubal factor infertility. RESULTS: The concentration of immunoreactive TNF alpha in FF was 0.36 +/- 0.02 microgram/L, and there were no significant correlations between levels of TNF alpha and steroids or FF volume. Bioactivity for TNF alpha was considerably less. Immunoreactive or bioactive TNF alpha was not detected in conditioned media from granulosa-lutein cell culture. Recombinant human TNF alpha dose-dependently stimulated proliferation of cultured granulosa-lutein cells as measured by incorporation of 3H-thymidine, but in contrast to earlier reports, we were not able to demonstrate any effect of rTNF alpha on basal or human chorionic gonadotropin-stimulated P accumulation during culture periods of up to 72 hours. The accumulation of both PGE2 and PGF2 alpha was dose-dependently increased by rTNF alpha during a 48-hour incubation period. Time course studies revealed that maximal levels of both PGE2 and PGF2 alpha were reached within 12 hours of culture. CONCLUSION: Immunoreactive and bioactive TNF alpha is present in FF. Tumor necrosis factor alpha may have a physiological role in stimulating proliferation of follicular cells and PG production at the time of ovulation and formation of the corpus luteum.

Ancistrobertsonine A and related naphthylisoquinoline alkaloids from Ancistrocladus robertsoniorum.

From the as yet unexplored East African Liana, Ancistrocladus robertsoniorum, several naphthylisoquinoline alkaloids have been isolated, based mainly on High Speed Countercurrent Chromatography (HSCCC). The structure of the new compound, ancistrobertsonine A, was elucidated by chemical and spectroscopic methods. Furthermore, the known alkaloids ancistrocladine, its atropo-diastereomer, hamatine, and its regioisomer, ancistrobrevine B, were isolated.