Differentiation of the mononuclear phagocyte system during mouse embryogenesis: The role of transcription factor PU.1

During mouse embryogenesis, macrophage-like cells arise first in the yolk sac and are produced subsequently in the liver. The onset of liver hematopoiesis is associated with the transition from primitive to definitive erythrocyte production. This report addresses the hypothesis that a similar transition in phenotype occurs in myelopoiesis. We have used whole mount in situ hybridization to detect macrophage-specific genes expressed during mouse development. The mouse c-fms mRNA, encoding the receptor for macrophage colony-stimulating factor (CSF-1), was expressed on phagocytic cells in the yolk sac and throughout the embryo before the onset of liver hematopoiesis, Similar cells were detected using the mannose receptor, the complement receptor (CR3), or the Microphthalmia transcription factor (MITF) as mRNA markers. By contrast, other markers including the F4/80 antigen, the macrophage scavenger receptor, the S-100 proteins, S100A8 and S100A9, and the secretory product lysozyme appeared later in development and appeared restricted to only a subset of c-fms-positive cells. Two-color immunolabeling on disaggregated cells confirmed that CR3 and c-fms proteins are expressed on the same cells. Among the genes appearing later in development was the macrophage-restricted transcription factor, PU.1, which has been shown to be required for normal adult myelopoiesis. Mice with null mutations in PU.1 had normal numbers of c-fms-positive phagocytes at 11.5dpc. PU.1(-/-) embryonic stem cells were able to give rise to macrophagelike cells after cultivation in vitro. The results support previous evidence that yolk sac-derived fetal phagocytes are functionally distinct from those arising in the liver and develop via a different pathway. (C) 1999 by The American Society of Hematology.

A review of the Apocreadiidae Skrjabin, 1942 (Trematoda : Digenea) and description of Australian species

The Apocreadiidae is reviewed and is considered to include genera recognised previously within the families Apocreadiidae, Homalometridae, Schistorchiidae, Sphincterostomatidae and Trematobrienidae. Key features of the family are extensive vitelline follicles, eye-spot pigment dispersed in forebody, I-shaped excretory vesicle, no cirrus-sac and genital pore opening immediately anterior to the ventral sucker (usually) or immediately posterior to it (Postporus Manter, 1949). Three subfamilies and 18 genera are recognised within the Apocreadiidae. The Apocreadiinae comprises Homalometron Stafford, 1904 (new syn. Barbulostomum Ramsey, 1965), Callohelmis n. g., Choanodera Manter, 1940, Crassicutis Manter, 1936, Dactylotrema Bravo-Hollis & Manter, 1957, Marsupioacetabulum Yamaguti, 1952, Microcreadium Simer, 1929, Myzotus Manter, 1940, Neoapocreadium Siddiqi & Cable, 1960, Neomegasolena Siddiqi & Cable, 1960, Pancreadium Manter, 1954, Procaudotestis Szidat, 1954 and Trematobrien Dollfus, 1950. The Schistorchiinae comprises Schistorchis Luhe, 1906, Sphincterostoma Yamaguti, 1937, Sphincteristomum Oshmarin, Mamaev & Parukhin, 1961 and Megacreadium Nagaty, 1956. The Postporinae comprises only Postporus. A key to subfamilies and genera of the Apocreadiidae is provided. It is argued that there is no convincing basis for the recognition of the genus Apocreadium Manter, 1937 and all its constituent species are combined with Homalometron. The following new combinations are proposed for species previously recognised within Apocreadium: Homalometron balistis (Manter, 1947), H. caballeroi (Bravo-Hollis, 1953), H. cryptum (Overstreet, 1969), H. longisinosum (Manter, 1937), H. manteri (Overstreet, 1970), H. mexicanum (Manter, 1937) and H. vinodae (Ahmad, 1985). Apocreadium uroproctoferum Sogandares-Bernal, 1959 is found to lack a uroproct and is made a synonym of H. mexicanum. Homalometron verrunculi nom. nov. is proposed to replace the secondarily pre-occupied H. caballeroi Lamothe-Argumedo, 1965. Barbulostomum is made a synonym of Homalometron and H. cupuloris (Ramsey, 1965) n. comb. is proposed. Neochoanodera is made a synonym of Choanodera and Choanodera ghanensis (Fischthal & Thomas, 1970) n. comb. is proposed. Species within the Apocreadiinae and Postporinae are reviewed and the following are recorded or described from Australian fishes: Homalometron wrightae n. sp. from Achlyopa nigra (Macleay), H. synagris (Yamaguti, 1953) n. comb. from Scolopsis monogramma (Cuvier), H. stradbrokensis n. sp. from Gerres subfasciatus Cuvier, Marsupioacetabulum opallioderma n. sp. from G. subfasciatus, Neoapocreadium karwarensis (Hafeezullah, 1970) n. comb. from G. subfasciatus, N. splendens n. sp. from S. monogramma and Callohelmis pichelinae n. g., n. sp. from Hemigymnus melapterus (Bloch), H. fasciatus (Bloch), Stethojulis bandanensis (Bleeker) andChoerodon venustus (De Vis). Callohelmis is recognised by the combination of absence of tegumental spines, caeca terminating midway between the testes and posterior end of body, ventral sucker enclosed in a tegumental pouch, prominent muscles radiating through the body from the ventral sucker, vitelline follicles not extending into the forebody, and a very short excretory vesicle that opens ventrally. New combinations for species previously recognised within Crassicutis are proposed as follows: Neoapocreadium caranxi (Bilqees, 1976) n. comb., N. gerridis (Nahhas & Cable, 1964) n. comb., N. imtiazi (Ahmad, 1984) n. comb. and N. marina (Manter, 1947) n. comb. The host-specificity and zoogeography of the Apocreadiinae are considered.

The genus Deretrema Linton, 1910 (Digenea : Zoogonidae) from southern Great Barrier Reef fishes, with a description of Deretrema woolcockae n. sp.

Two species of Deretrema (Zoogonidae) are reported from labrid fishes from the Great Barrier Reef. D. nahaense Yamaguti, 1942 is recorded from the gall-bladders of the labrids Thalassoma hardwicke (Bennett), T. jansenii (Bleeker), T. lunare (Linnaeus) and T. lutescens (Lay & Bennett). This species is recognised, despite having been formerly synonymised with D. pacificum Yamaguti, 1942. In addition to morphological distinction, D. nahaense appears to have strict host-specificity for the genus Thalassoma. D. woolcockae n.sp. is described from the gall-bladder of Hemigymnus fasciatus (Bloch). The new species is close to D. acutum Pritchard, 1963 and D. plotosi Yamaguti, 1940, but differs slightly in the distribution of the vitelline follicles, the sucker-ratio and the position of the cirrus-sac. In addition, this species also appears to have a distinct host-specificity, being restricted to one labrid species.

The status of the genera Hysterolecithoides Yamaguti, 1934, Neotheletrum Gibson & Bray, 1979 and Machidatrema Leon-Regagnon, 1998 (Digenea : Hemiuroidea), including a description of M. leonae n. sp from Australian waters

A diagnosis is given for the lecithasterid genus Hysterolecithoides Yamaguti, 1934, which is now found to have two to six (possibly seven) vitelline masses. The species H. frontilatus (Manter, 1969) is returned to the genus, having been considered a member of the bunocotylid genus Neotheletrum by recent authors. It is redescribed from Siganus nebulosus, Moreton Bay, and S. doliatus, Lizard Island, Great Barrier Reef and New Caledonia, with emphasis on the presence of Juel's organ, a uterine seminal receptacle and the blind sac associated with the genital atrium. It differs from its congeners in the trajectory of the pars prostatica which recurves dorsally to the sinus-sac. Oligolecithoides Shen, 1982 is synonymised with Hysterolecithoides and O. trilobatus Shen, 1982 is synomised with H. epinepheli Yamaguti, 1934. Machidatrema Leon-Regagnon, 1998 is diagnosed, and found to be close to Hysterolecithoides, but differs in the lack of a blind-sac projecting from the dorsal genital atrium, by its tandem testes, the coiling of the uterus between the testes and the ovary, and the ventral excretory pore. M. leonae n. sp. is described from Siganus fuscescens, S. lineatus, S. doliatus, S. corallinus, S. vulpinus and Scarus globiceps at Heron Island, Queensland. It differs from its closest congener, M. akeh, in the muscular and tegumental flap over the genital pore and details of the terminal genitalia. M. chilostoma (Machida, 1980) and M. kyphosi (Yamaguti, 1970) are redescribed from Kyphosus vaigiensis from Heron Island. Neotheletrum Gibson & Bray, 1979 is diagnosed: it differs from Hysterolecithoides in its confluent excretory arms, blind seminal receptacle (no Juel's organ) and uniformly tripartite vitellarium. A cladistic analysis suggests that M. chilostoma and M. kyphosi are not best accommodated in Machidatrema, that Machidatrema (sensu stricto) is monophyletic and that Hysterolecithoides is paraphyletic. Hysterolecithoides and Machidatrema are considered hysterolecithine lecithasterids, whilst Neotheletrum is retained as an opisthadenine bunocotylid.

The status of Petalocotyle Ozaki, 1934 (Digenea : Gyliauchenidae), including the description of two new species from acanthurid fishes in Queensland, Australia

The status of Petalocotyle Ozaki, 1934 within the Gyliauchenidae Goto & Matsudaira, 1918 is reviewed. Two new species, P. adenometra from Prionurus microlepidotus (Amity Point, Queensland, Australia) and P. diverticulata from Acanthurus nigrofuscus and A. lineatus (Heron Island, Queensland, Australia), are described. The body plan of Petalocotyle conforms to that of members of the Gyliauchenidae (oral sucker absent, well-developed pharynx, complex oesophagus and characteristic male terminal genitalia), indicating justifiable inclusion in this family. A new diagnosis is given for the genus, such that Petalocotyle is now identified by the presence of an anterior, protuberant ventral sucker, long caeca, a large, sigmoid cirrus-sac containing a coiled ejaculatory duct, and an extensive vitellarium. We suggest that, of all the known genera of gyliauchenids, Petalocotyle may most closely resemble the 'archaetypal gyliauchenid', that is, it may be placed basally within the radiation of the Gyliauchenidae. However, derived characters, like diverticula in the reproductive system, indicate that some characters of individual members of Petalocotyle may be considered advanced and do not reflect an archaetypal condition. Parallels in the structure of the male and female genitalia of Robphildollfusium Paggi & Orecchia, 1963 and Petalocotyle, along with the shared morphology of the digestive tract, indicate possible phylogenetic links between the two genera. This affinity is difficult to infer using morphology alone and recommend that Robphildollfusium remain detached from the Gyliauchenidae.

Scorpidotrema longistipes n. g., n. sp. (Digenea: Opecoelidae) from Scorpis georgiana (Teleostei: Scorpididae) from southern Western Australia

Scorpidotrema longistipes n. g., n. sp. is described from the intestine of Scorpis georgiana Valenciennes (Scorpididae) from off Point Peron, Western Australia. The new genus is distinguished by the combination of a remarkably long and retractable ventral sucker peduncle, a possible uroproct, well-developed cirrus-sac and a uterine seminal receptacle. The subfamilial relationships of the new genus are troublesome. It incorporates features of the Opecoelinae, Stenakrinae and Plagioporinae. The absence of a canalicular seminal receptacle suggests a relationship with the Opecoelinae and Stenakrinae, whereas the well-developed cirrus-sac suggests a relationship with the Plagioporinae and Stenakrinae. The overall arrangement of the gonads is not similar to that of existing genera of Stenakrinae. It is concluded that the genus is best placed in the Stenakrinae although that subfamily may now be an artificial assemblage. This new genus forms part of a distinctive fauna of trematodes restricted to Australian southern temperate fishes.

Metformin therapy and diabetes in pregnancy

No adverse pregnancy outcomes with metformin use have been reported, except in one unmatched study. Otherwise, the studies are small and non-randomised, with the exception of one prospective, randomised controlled trial, currently under way, comparing metformin with insulin in women with gestational diabetes mellitus (the MiG trial). No long-term follow-up data for offspring of mothers receiving metformin have been published. Any woman with diabetes should be as close to euglycaemia as possible before pregnancy. In some circumstances (eg, severe insulin resistance), metformin therapy during pregnancy may be warranted. When metformin treatment is being considered, the individual risks and benefits need to be discussed with the patient so that an appropriate decision can be reached.

Successful outcome of a surgically treated giant retroperitoneal liposarcoma during pregnancy

Background Retroperitoneal liposarcomas occur more frequently between the fifth and seventh decades. Fortunately, these tumors are exceedingly rare in pregnancy, but when they occur, their management becomes even more challenging. Case A pregnant patient with a retroperitoneal liposarcoma was treated by complete surgical resection at 13 weeks of gestation. The patient gave birth at 37 weeks of gestation to a normal newborn and remains free of disease after a year. Conclusions Individualized approach should be taken according to gestational phase, tumor kinetics and overall patient`s condition. Any disturbance to materno-fetal equilibrium, should be an indication for urgent therapeutic approach.

Cardiac troponin T as a biochemical marker of cardiac dysfunction and ductus venosus Doppler velocimetry

Objectives: The aim of this study was to determine the correlation between ductus venosus (DV) Doppler velocimetry and fetal cardiac troponin T (cTnT). Study design: Between March 2007 and March 2008, 89 high-risk pregnancies were prospectively studied. All patients delivered by cesarean section and the Doppler exams were performed on the same day. Multiple regression included the following variables: maternial age, parity, hypertension, diabetes, gestational age at delivery, umbilical artery (UA) S/D ratio, diagnosis of absent or reversed end-diastolic flow velocity (AREDV) in the UA, middle cerebral artery (MCA) pulsatility index (131), and DV pulsatility index for veins (PIV). Immediately after delivery, UA blood samples were obtained for the measurement of pH and cTnT levels. Statistical analysis included the Kruskal-Wallis test and multiple regressions. Results: The results showed a cTnT concentration at birth >0.05 ng/ml in nine (81.8%) of AREDV cases, a proportion significantly higher than that observed in normal UA S/D ratio and UA S/D ratio >p95 with positive diastolic blood flow (7.7 and 23.1%, respectively, p < 0.001). A positive correlation Was found between abnormal DV-PIV and elevated cTnT levels in the UA. Multiple regression identified DV-PIV and a diagnosis of AREDV as independent factors associated with abnormal fetal cTnT levels (p < 0.0001, F(2.86) = 63.5, R = 0.7722). Conclusion: DV-PIV was significantly correlated with fetal cTnT concentrations at delivery. AREDV and abnormal DV flow represent severe cardiac compromise, with increased systemic venous pressure, and a rise in right ventricular afterload, demonstrated by myocardial damage and elevated fetal cTnT. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

Growth Hormone Receptor Exon 3 Isoforms and Their Implication in Growth Disorders and Treatment

Human recombinant growth hormone (hGH) has been used to treat short stature in several different conditions, but considerable inter-individual variation in short- and long-term growth response exists. Pharmacogenomics can provide important insights into hGH therapy. The GH receptor (GHR) is the first key molecule mediating GH action. In the past 3 years, a common GHR polymorphism reflecting the presence (GHRf1) or absence (GHRd3) of exon 3 has been under intensive investigation regarding its influence on the response to hGH therapy. Studies that evaluated response to GH treatment determined by these two GHR isoforms in children with GH deficiency, girls with Turner syndrome, children born small for gestational age and patients with acromegaly showed that patients carrying the GHRd3 allele demonstrated a greater GH sensitivity than patients homozygous for the GHRf1 allele. Other studies presented contradictory data, however, which may be caused by confounding factors such as small sample sizes and differences in experimental design. This GHR exon 3 genotype is the first identified genetic factor found to modulate the individual response to GH therapy. This article reviews the historical aspects and pharmacogenetic studies published to date in relation to this GHR polymorphism. The analyses of present and future validation studies may define the use of this and other polymorphisms in clinical practice, moving from pharmacogenetics to routine application and allowing individualization of hGH doses to optimize final outcome. Copyright (C) 2009 S. Karger AG, Basel

Morphometric Study on the Anatomy of the Fetal Cricoid Cartilage and Comparison Between Its Inner Diameter and Endotracheal Tube Sizes

Objectives: The high incidence of respiratory disorders is one of the main problems in perinatal medical care. With the increased use of intubation, the incidence of laryngeal injury causing stenosis has also increased. The principal constriction point in the infant`s larynx is the midcricoid area. We sought to provide detailed morphometric data on the anatomy of the cricoid cartilage and its relationship with growth and body characteristics of fetuses at 5 to 9 months of gestational age. Methods: Nineteen larynges obtained from 17 stillborn infants and 2 newborn infants ranging in gestational age from 5 to 9 months were studied. Measurements of the cricoid cartilage were made with a millimeter-graded caliper. Results: Weight was the variable most correlated with cricoid measurements. The cricoid lumen configuration showed an almost elliptic shape and did not change with gestational age. The mean inner subglottic cricoid area was 19.27 +/- 9.62 mm(2) and was related to weight and body surface area. Cricoid growth was more pronounced at the outer portion of the cartilage. Conclusions: The cricoid lumen configuration was elliptic, and its mean area was smaller than that of available endotracheal tubes. This lumen area was most influenced by weight and height.

Subpopulations of corticotrophs in the sheep pituitary during late gestation: Effects of development and placental restriction

The prepartum surge in fetal plasma cortisol is essential for the normal timing of parturition in sheep and may result from an increase in the ratio of ACTH to proopiomelanocortin (POMC) in the fetal circulation. In fetuses subjected to experimental induction of placental restriction, the prepartum surge in fetal cortisol is exaggerated, whereas pituitary POMC mRNA levels are decreased, and in vitro, unstimulated ACTH secretion is elevated in corticotrophs nonresponsive to CRH. We therefore investigated the changes in the relative proportions of cells expressing POMC, ACTH, and the CRH type 1 receptor (CRHR1) shortly before birth and during chronic placental insufficiency. Placental restriction (PR) was induced by removal of the majority of placental attachment sites in five ewes before mating. Pituitaries were collected from control and PR fetal sheep at 140 d (control, n = 4; PR, n = 4) and 144 d (control, n = 6; PR, n = 4). Pituitary sections were labeled with specific antisera raised against POMC, ACTH, and CRHR1. Three major subpopulations of corticotrophs were identified that expressed POMC + ACTH + CRHR1, ACTH + CRHR1, or POMC only. The proportion of pituitary corticotrophs expressing POMC + ACTH + CRHR1 decreased (P < 0.05) between 140 (control, 60 +/- 1%; PR, 66 +/- 4%) and 144 (control, 45 +/- 2%; PR, 56 +/- 6%) d. A significantly higher (P < 0.05) proportion of corticotrophs expressed POMC + ACTH + CRHR1 in the pituitary of the PR group compared with controls. This study is the first to demonstrate subpopulations of corticotrophs in the fetal sheep pituitary that differentially express POMC, ACTH, and CRHR1 and the separate effects of gestational age and placental restriction on these subpopulations of corticotrophs.

Regulation of CSF-1 receptor expression

Cells of the mononuclear phagocyte lineage possess receptors for macrophage colony-stimulating factor (CSF-1) encoded by the c-fms protooncogene and respond to CSF-1 with increased survival, growth, differentiation, and reversible changes in function. The c-fms gene is itself a macrophage differentiation marker. In whole mount analyses of mRNA expression in embryos, c-fms is expressed at very high levels on placental trophoblasts. It is detectable on individual cells in the yolk sac around 8.5 to 9 days postcoitus, appears on isolated cells in the head of the embryo around 9.5 dpc, and appears on numerous cells throughout the embryo by day 10.5. The extent of c-fms expression is much greater than for other macrophage-specific genes including lysozyme and a macrophage-specific protein tyrosine phosphatase. Our studies of the cis-acting elements of the c-fms promoter have indicated a key role for collaboration between the macrophage-specific transcription factor, Pu.1, which functions in determining the site of transcription initiation, and other members of the Ets transcription factor family. This is emerging as a common pattern in macrophage-specific promoters. We have shown that two PU box elements alone can function as a macrophage-specific promoter. The activity of both the artifical promoter and the c-fms promoter is activated synergistically by coexpression of Pu.1 and another Ets factor, c-Ets-2. A 3.5kb c-fms exon 2 promoter (but not the 300bp proximal promoter) is also active in a wide diversity of tumor cell lines. The interesting exception is the melanoma cell line K1735, in which the promoter is completely shut down and expression of c-fms causes growth arrest and cell death. The activity of the exon 2 promoter in these nonmacrophages is at least as serum responsive as the classic serum-responsive promoter of the c-fos gene. It is further inducible in nonmacrophages by coexpression of the c-fms product. Unlike other CSF-1/c-fms-responsive promoters, the c-fms promoter is not responsive to activated Ras even when c-Ets-2 is coexpressed. In most lines, production of full length c-fms is prevented by a downstream intronic terminator, but in Lewis lung carcinoma, read-through does occur, and expression of both c-fms and other macrophage-specific genes such as lysozyme and urokinase becomes detectable in conditions of serum deprivation. (C) 1997 Wiley-Liss, Inc.

Postlepidapedon Zdzitowiecki, 1993 and Gibsonivermis n g (Digenea: Lepocreadiidae) from fishes of the southern Great Barrier Reef, Australia, and their relationship to Intusatrium Durio & Manter, 1968

The genus Intusatrium Durio & Manter, 1968 is redefined based on a re-examination of paratypes of the type-species, I. robustum Durio & Manter, 1968, and is considered monotypic with characteristic terminal genitalia: internal seminal vesicle elongate tubular, with rather thick wall, divided by slight change in wall thickness into longer proximal and shorter distal region; pars prostatica subcylindrical; ejaculatory duct relatively short, with wrinkled/wall. The genus Postlepidapedon Zdzitowiecki, 1993 is redefined and Intusatrium secundum Durio & Manter, 1968 is attributed to it as a new combination. Postlepidapedon secundum n. comb. is redescribed from a paratype and new material from Choerodon graphicus. P. spissum n. sp. from Choerodon venustus, C. cyanodus, C. fasciatus and C. schoenleinii is recognised on the basis of its thick-walled internal seminal vesicle. I! uberis n. sp. from Choerodon schoenleinii and C. venustus is distinguished by the shape and contents of the cirrus-sac with narrow, convoluted internal seminal vesicle, large vesicular pars prostatica and short, muscular ejaculatory duct. A new genus, Gibsonivermis, erected for Intusatrium berryi Gibson, 1987, is characterised by the elongate narrow cirrus-sac and a uroproct. G. berryi n. comb. is redescribed from Sillago ciliata, S. maculata and Sillago sp.

Growth hormone-binding protein in normal and pathologic gestation: Correlations with maternal diabetes and fetal growth

To date, measurements of GH-binding protein (GHBP) during human pregnancy have been carried out using;assays susceptible to interference by the elevated levels of human placental GH typical of late gestation. We recruited a large cohort of pregnant women (n = 140) for serial measurements of GHBP and used the ligand immunofunctional assay for GHBP. For normal gravidas, GHBP levels fell throughout gestation. Mean levels were 1.07 nmol/L (SE = 0.18) in the first trimester, 0.90 nmol/L (SE = 0.08) at 18-20 weeks, 0.73 nmol/L (SE = 0.05) at 28-30 weeks, and 0.62 nmol/L (SE = 0.06) at 36-38 weeks. GHBP levels in the first trimester correlated significantly with maternal body mass index (r = 0.58; P < 0.01). GHBP levels in pregnancies complicated by noninsulin-dependent diabetes mellitus (NIDDM) were substantially elevated at all gestational ages. The mean value in the first quarter (2.29 nmol/L) was more than double the normal mean (P < 0.01). In contrast, patients with insulin-dependent diabetes mellitus (IDDM) showed reduced GHBP concentrations at 36-38 weeks. The correlation between body mass index and GHBP is consistent with a metabolic role for GHBP during pregnancy, as is the dramatic elevation in GHBP observed in cases of NIDDM. At 36 weeks gestation, GHBP was significantly elevated (P < 0.01) in those women whose neonates had low birth weight (