bluuebunny/crossref_metadata_2025 huggingface dataset:
Fetal pig islet xenografts in NOD/Lt mice: The effect of peritransplant anti‐CD4 monoclonal antibody and graft immunomodification on graft survival, and lack of expression of Gal(α1–3)Gal on endocrine cells
Authors: Koulmanda Maria , McKenzie I.F.C. , Sandrin M.S. , Mandel T.E.
Published: November 2008

Abstract: Transplantation of tissues that are vascularized by ingrowth of host vessels, as opposed to primarily vascularized grafts, may be one way of avoiding hyperacute rejection (HAR). Pretransplant treatment of the graft to eliminate from it highly immunogenic cells may also improve graft survival. We examined organ cultured fetal pig pancreas in prediabetic NOD/Lt female mice treated with peritransplant anti‐CD4 MAb (GK 1.5) and in separate experiments also studied the expression of Gal(α 1–3)Gal, an epitope that may be a major target of a human anti‐pig response. Immunocytochemistry to detect insulin, glucagon, and somatostatin and Gal(α 1–3)Gal showed that differentiated (i.e., hormone containing) endocrine cells were Gal(α 1–3)Gal negative but hormone‐negative ductal cells showed strong Gal(α1–3)Gal staining on their luminal border and interstitial cells were also positive particularly early after transplantation. Rejection occurred in all animals, but its rate was markedly altered by transient immunosuppression with the depleting anti‐CD4 MAb. Cell infiltration was first detected in controls 2 days posttransplant, was pronounced by 4 days, and grafts showed advanced rejection by 7 days whether or not they had been “immunomodified” by 2 days exposure to 90% O2. In contrast, peritransplant immunosuppression with GK 1.5 improved graft survival but rejection was advanced by 28 days. The control graft sites had many eosinophils, macrophages and mast cells by 4 days, but little infiltration was seen until after 14 days in the immunosuppressed mice and was predominantly by mononuclear cells. Use of “immunomodified” grafts was of no benefit. Thus, graft immunomodification per se, that often allows islet allografts to survive in rodents, had no effect on xenograft survival in this model. A major potential target antigen for natural Ab in humans, Gal(α 1–3)Gal, is not present on differentiated graft endocrine cells.
Recognition of an octapeptide sequence by multiple Galα(1,3)Gal‐binding proteins
Authors: Vaughan Hilary A. , Oldenburg Kevin R. , Gallop Mark A. , Atkin Julie D. , McKenzie Ian F.C. , Sandrin Mauro S.
Published: November 2008

Abstract: Strategies are now being designed to overcome the hyperacute rejection of vascularized pig xenografts by human natural anti‐Galα(1,3)Gal antibodies. We now demonstrate that a synthetic octapeptide, Gal pep 1 (DAHWESWL), isolated from a peptide epitope library using the α‐galactosyl specific lectin IB4, “mimics” the carbohydrate epitope Galα(1,3)Gal. In vitro studies demonstrated that the binding of the IB4 lectin or human natural antibodies to pig cells could be specifically blocked by Gal pep 1, in several different serological assays: a) the hemagglutnation of pig erythrocytes; b) cytofluorographic analysis of pig lymphocytes and endothelial cells; c) cytotoxicity of human serum against pig endothelial cells; d) an ELISA assay. The relative affinity of the peptide for the IB4 lectin was similar to that of α‐galactosyl sugars, although it was bound at a lower affinity by human antibodies. The implications of these observations to xenotransplantation are that peptides could theoretically be used to block hyperacute rejection.
Protection from complement‐mediated injury in livers and kidneys of transgenic mice expressing human complement regulators
Authors: Mora Marirosa , Mulder Lubbertus C.F. , Lazzeri Massimo , Boschi Manuela , Ciccopiedi Eugenia , Melli Cristina M. , Bruzzone Paolo , Alfani Dario , Cortesini Raffaello , Rossini Mara
Published: November 2008

Abstract: The major problem in the use of phylogenetically distant donors is a fast, strong reaction called hyperacute rejection. This reaction mediated by complement is directed against the vascular endothelia of the transplanted organ. Complement activation is tightly controlled by several regulatory proteins which inhibit the formation and function of different complement components. To verify the hypothesis that organs expressing such inhibitory factors could be spared from complement‐mediated hyperacute rejection, we have generated mice transgenic for the human complement inhibitor membrane cofactor protein (hMCP) and decay accelerating factor (hDAF). Different levels of hMCP and/or hDAF expression, according to the promoter used, were detected by RNA analysis in the major organs, specifically on the organ vascular endothelia, as revealed by immunohistochemical analysis. The development of an in vivo model of human plasma perfusion allowed the characterization of complement‐mediated damage in control animals and the degree of protection due to the presence of hMCP, hDAF, or both in the organs derived from single or double transgenic mice. In this paper we compare the level of expression of complement regulators with the degree of protection in two major organs: liver and kidney.
Pediatric cardiac xenograft growth in a rhesus monkey‐to‐baboon transplantation model
Authors: Matsumiya Goro , Gundry Steven R. , Fukushima Norihide , Kawauchi Motohiro , Zuppan Craig W. , Bailey Leonard L.
Published: November 2008

Abstract: It is unclear that organs from other species could grow successfully in the environment of another species. We evaluated the growth capacity of a cardiac xenograft in a rhesus monkey‐to‐juvenile baboon orthotopic heart transplantation model where there exists a discrepancy of the organ size and its growth rate between donor and recipient species. Eight recipient baboons who survived over 60 days were followed for body weight (BW) and the size of xenografts (left ventricular end‐diastolic volume, LVEDV; left ventricular mass, LVM; and left ventricular posterior wall thickness, LVPWT) by echocardiography at 1 week and monthly after xenotransplantation. One baboon is growing along the normal growth curve. The other three baboons gained BW in parallel with normal growth curve except for a few months after surgery. LVEDV, LVM, and LVPWT increased constantly in these four baboons. In one baboon, BW increased over 1 year to 5.5 kg (which was almost the same as BW of adult rhesus monkey) and then plateaued. LVEDV, LVM, and LVPWT increased until 1 year posttransplantation, but then stopped when BW plateaued. In the other four baboons who did not gain BW because of infectious complications or side effects by immunosuppressive therapy, LVEDV, LVM, and LVPWT did not change significantly. We conclude that a cardiac xenograft from a rhesus monkey can successfully grow and support normal growth of a juvenile baboon, but whether a recipient can grow beyond the donor size is yet unknown.
Introduction of α(1,2)‐fucosyltransferase and its effect on a‐Gal epitopes in transgenic pig
Authors: Koike Chihiro , Kannagi Reiji , Takuma Yoshihiro , Akutsu Fumiko , Hayashi Satoshi , Hiraiwa Nozomu , Kadomatsu Kenji , Muramatsu Takashi , Yamakawa Hirohito , Nagai Takuya , Kobayashi Shoji , Okada Hidechika , Nakashima Izumi , Uchida Kazuharu , Yokoyama Itsuo , Takagi Hiroshi
Published: November 2008

Abstract: Hyperacute rejection (from pig to human) is thought to result from activation of complement initiated by the binding of host natural antibodies to α‐galactosyl (α‐Gal) epitopes of donor endothelial cells. However, α‐Gal epitope shares a common precursor with H antigen in humans. This means that H antigens as well as α‐Gal epitopes are synthesized in a competitive manner by different enzymes. We thought that it would be possible to convert α‐Gal epitopes into H antigens by introducing cDNA of α(1,2)‐fucosyltransferase (α1–2FT) into porcine cells, and so, pig embryos were microinjected with αl‐2FT cDNA. Transgenic pigs that carried α1–2FT were thus established. Cytotoxicity of fibrocytes derived from skin of transgenic pig was measured by 51Cr release assay, which showed that H antigen‐expressing cells were significantly resistant to a challenge with human sera. These experiments indicate that our method provides a new strategy which contributes to a successful discordant xenotransplantation.
Reduction of the major porcine xenoantigen Galoc(1,3)Gal by expression of α(1,2)fucosyltransferase
Authors: Sandrin Mauro S. , Fodor William L. , Cohney Shlomo , Mouhtouris Effie , Osman Narin , Rollins Scott A. , Squinto Stephen P. , McKenzie Ian F. C.
Published: November 2008

Abstract: Although removal of the Galα(1,3)Gal antigen from pigs would prevent hyperacute graft rejection, the technique of homologous recombination to knock out the α 1,3 galactosyltransferase gene is not available for pigs, and an alternative strategy is presented. As both α 1,3 galactosyltransferase and α 1,2 fucosyltransferase use the same substrate (N‐acetyl lacto‐samine), competition between the transferases in vitro and in vivo was examined. The data show that there is indeed a hierarchy of these gly‐cosyltransferases competing for the same substrate, and that α 1,2 fuco‐syltransferase takes precedence over α 1,3 galactosyltransferase: a) COS cells simultaneously transfected with cDNA clones encoding α, 2 fuco‐syltransferase and α 1,3 galactosyltransferase show preferential expression of the H substance (synthesised by α 1,2fucosyltransferase) rather than Galα(1,3)Gal (synthesised by α 1,3galactosyltransferase), even though α 1,3galactosyltransferase mRNA and functional enzyme was present, b) In a pig kidney cell line that expressed both the Galα(1,3)Gal and H, the increased expression of H induced by the transfection and stable expression of α 1,2fucosyltransferase resulted in decreased expression of Galα(1,3)Gal. c) Coexpression of α 1,2fucosyltransferase and α 1,3galactosyltransferase in either COS cells or the pig cell line resulted in decreased human antibody binding and complement‐mediated cell lysis, d) Transgenic mice, ubiquitously expressing α 1,2fucosyltransferase show a major decrease in Galα‐(1,3)Gal expression and a decrease in natural human antibody binding. These findings have important implications for xenotransplantation in that α,2fucosyltransferase transgenic pigs could be a source of donors for xenotransplantation to humans.
Direct activation of human responder T cells by porcine stimulator cells leads to T cell proliferation and cytotoxic T cell development
Authors: Satake M. , Kawagishi N. , Möller E.
Published: November 2008

Abstract:Functional activities of highly purified T human responder lymphocytes reactive against porcine stimulator cells were studied to evaluate whether porcine stimulator cells can directly activate a xenospecific cellular immune response. Mixed lymphocyte culture (MLC) tests revealed that CD4+ human responder cells proliferate when stimulated in vitro with porcine cells, a reaction similar to that of alloresponses regarding magnitude and tempo. A direct pathway of activation was verified by the requirement for adherent porcine stimulator cells and the partial blocking by monoclonal antibodies against porcine major histocompatibility complex (MHC) class II antigens. An analysis of proliferative CD4+, CD3+, CD16‐, and 56‐ human T cell clones revealed that some clones were seemingly recognizing allele‐specific determinants, whereas others could be restimulated by a wide range of porcine stimulator cells. Cytotoxic T cells (CTLs) were generated following direct recognition of pig cell ligands by human T cells in the absence of autologous antigen‐presenting cells (APCs). Although the identification of target antigen(s) on the pig cell recognized by the CTL warrants some discussion, the pattern of killing exhibited by the CTLs indicates the recognition of porcine polymorphic determinant(s). The implications of these findings for cellular reactivity against porcine transplants are discussed.
Protection of guinea pig cells from rat complement‐mediated lysis by the transfection of rat complement regulatory factor 512 gene
Authors: Hayashi Shuji , Okada Hidechika , Emi Nobuhiko , Okada Noriko , Yokoyama Itsuo , Takagi Hiroshi
Published: November 2008

Abstract: 512 antigen, rat complement regulatory factor, has been reported to possess the same structure as Crry/p65, mouse complement regulatory protein, which functions like decay‐accelerating factor (DAF) and/or membrane cofactor protein (MCP). This study was designed to evaluate the Nagoya, Japan protective effect on guinea pig cells transfected with 512 cDNA from rat complement mediated lysis. 512 antigen was expressed on rat cells alone, not on guinea pig, pig, or human cells. Guinea pig cells transfected with 512 cDNA were protected from rat complement mediated injury through the inhibition of C3 activation. The magnitude of injury was inversely proportional to the intensity of 512 antigen expression, although the protective effect was not always species‐specific. These results demonstrate that the genetic engineering to guinea pig cells with 512 cDNA is effective for the inhibition of rat complement activation.
Mechanisms of hyperacute rejection of discordant liver xenograft
Authors: Hengster P. , Lsnke R. , Feichtinger J. , Hechenleitner P. , Mark W. , Eberl T. , Klima G. , Huemer H. , Daha M. , Margreiter R.
Published: November 2008

Abstract: Use of discordant liver xenografts is a possible means of overcoming the ever‐increasing organ shortage. Immunological barriers, however, remain the major obstacle to the application of this therapeutic option. The aim of the study was to establish the model and to further investigate mechanisms of hitherto assumed liver xenograft rejection in the discordant guinea pig to rat combination and to determine the role of complement activation in this model.

Male guinea pigs served as donors, and all recipient Lewis rats underwent splenectomy prior to orthotopic liver transplantation with a modified cuff technique. Animals were grouped according to their pharmacologic therapy: group 1: control group, no further therapy; group 2: plasmapheresis immediately prior to transplantation; group 3: Cobra venom factor; group 4: plasmapheresis and Cobra venom factor; group 5: Cyclosporin A; and group 6: RS 61443 with three animals in each group. Median survival was 62 (52–68) minutes in group 1, 150 (129–190) in group 2, 745 (260–1034) in group 3, 45 (18–60) in group 4, 91 (76–118) in group 5, and 212 (170–245) in group 6. Histologically, portal and sinusoidal congestion, interstitial haemorrhages, intragraft granulocyte deposits, and parenchymal damage were found to be most pronounced in groups 1 and 3. In all groups endothelial cells stained positive for IgM and complement component C3 compatible with classical pathway activation. From our findings where deposition of Factor H was noted it is concluded that in this discordant model the complement system is activated by the alternative pathway. Intragraft IgM deposits, however, suggest that the classical activation pathway may also be involved.
Protective genes expressed in endothelial cells of second hamster heart transplants to rats carrying an accommodated first graft
Authors: Hechenleitner Paul , Mark Walter , Candinas Daniel , Miyatake Tsukasa , Koyamada Nozomi , Hancock Wayne W. , Bach Fritz H.
Published: November 2008

Abstract: Accommodation refers to survival of a xenograft despite the presence of anti‐donor organ antibodies and complement. We have recently shown that accommodation of a hamster heart transplanted to a rat receiving short‐term cobra venom factor (CVF) and continuing cyclosporine A (CyA) therapy is associated with i) the expression in the endothelial cells (EC) and smooth muscle cells of the graft of a number of “protective” genes, ii) a prominent intragraft Th2 cytokine profile, and iii) the relatively heavy deposition of IgG2c antibodies on the EC of the graft. In contrast, rejecting grafts do not express the protective genes, have a Th1 cytokine profile, and apparently have lesser amounts of IgG2c. These findings are consistent with host factors (Th2 cytokines and IgG2c) contributing to xenograft accommodation. To test whether these host factors may predispose to the development of accommodation, we placed a second hamster heart into each of 12 rats carrying a surviving first heart; recipients were, at the time, receiving only CyA. Whereas first grafts transplanted to rats receiving only CyA survive for 3 to 4 days, 11 out of 12 second transplants survived more than 20 days, and the other survived for 7 days. Nine of the twelve were not rejected: of these, four were removed between day 35 and 132 for study, and the remainder are still beating at 35 to 52 days. The surviving second hearts we studied had accommodated in that the picture on immunopathology was the same as for surviving first hearts. We suggest that the Th2 cytokines and perhaps the IgG2c response are factors in allowing prolonged survival of the second grafts and, further, that these factors contribute to the expression in the EC and smooth muscle cells of the surviving second hearts of the protective genes.
Development of thymus autografts under the kidney capsule in the pig: A new “organ” for xenotransplantation
Authors: Lambrigts D. , Franssen C. , Martens H. , van Calster P. , Meurisse M. , Geenen V. , Charlet‐Renard C. , Dewaele A. , Coignoul F. , Lamy M. , Alexandre G.P.J.
Published: November 2008

Abstract:Ten piglets, 7 to 16 weeks old, were partially thymectomised and 1 to 4 cm3of minced thymic fragments autografted under the renal capsule. They were sacrificed, respectively, after 2, 4, 6, 8, 12, and 20 weeks. After 2 weeks, irregular whitish zones are present under the renal capsule. They were composed principally of two cell types: the first type was characterized by small round basophilic nuclei and little cytoplasm typical of lymphocytes; the second cell type had larger ovoid nuclei and a large vacuolised cytoplasm. Each cell type could be found in separate lobules or mixed in variable proportion in the same structure. The thymic autografts grew to form a layer up to 4 mm thick after 20 weeks. In the meantime, at the beginning of 4th week, the lobular structure became well organized with the cell type presenting large nuclei and cytoplasm being restricted to the center of the lobules while lymphocytes composed a peripheral layer. Hassal corpuscles (HC) appeared in the center of the lobules. Immunohistochemical labeling with anti‐cytokeratin mono‐ and poly‐ clonal Ab and with anti‐neurophysin polyclonal Ab displayed all the characteristics of normal functional thymic microenvironment. It is proposed that this novel experimental preparation ending up as a neo‐organ (thymo‐kidney) be used for xenotransplantation in an attempt to produce specific xenotolerance.
Cellular rejection of fetal pancreas grafts: Differences between alio‐ and xenograft rejection
Authors: Mandel T. E. , Kovarik J. , Koulmanda M. , Georgiou H.M.
Published: November 2008

Abstract: Hyperacute rejection (HAR) is the major immunologic problem with vascularized xenografts between discordant donor/recipient combinations but does not occur in neovascularized grafts of organ‐cultured fetal pig pancreas in either mice or cynomolgus monkeys. However, a form of cell‐mediated acute rejection with quite different histopathologic features does occur with kinetics that are similar to acute cellular rejection of fetal pancreas allografts in non‐immunosuppressed MHC‐mismatched mice. Xenograft rejection is dominated by non‐lymphoid cells, mostly eosinophils, that appear some days after transplantation. In contrast, in mouse allografts, mononuclear cells are the dominant population throughout the rejection process^. The rejected allograft site rapidly resolves to form a mature non‐irifiltrated scar whereas the infiltrate in the xenograft site remains for weeks and forms a large granuloma before its eventual resolution. There are also differences in the intra‐graft cytokine profile in the graft site between alio‐ and xenografts during acute rejection with an early predominance of IL‐5 and TNF‐α and an absence of TNF‐γ in the xenografts. Immunosuppression with a depleting anti‐CD4 mAb shows that xenograft rejection is more dependent on CD4+ve T cells but xenografts are more difficult to maintain with conventional immunosuppression that is often effective for allografts. Limited studies in primates have shown that the histopathology of fetal pig pancreas rejection is similar to that seen in mice but occurs at a faster tempo. Thus, although HAR may not be a problem in rejection of neovascularised xenografts, a vigorous form of cellular rejection is present that may require different immunosuppression than is usually used for the I control of allograft rejection.
The role of monocytes and macrophages in delayed xenograft rejection
Authors: Fryer Jonathan P. , Chen Sally , Johnson Eric , Simone Pamela , Sun Lin Hong , Goswitz Joseph J. , Matas Arthur J.
Published: November 2008

Abstract: Despite the development of successful strategies for averting hyperacute rejection (HAR) in both small and large animal xenograft models, a delayed xenograft rejection (DXR) ultimately occurs. This process is characterized by endothelial cell activation and graft infiltration with activated monocytes and natural killer (NK) cells. We evaluated the role of monocytes and macrophages in a guinea pig‐to‐rat model of DXR. Our results suggest that specific interactions between these cells and the xenograft occur that result in their activation, since adoptive transfer of xenoactivated splenocytes significantly accelerated both DXR and allograft rejection, while adoptive transfer of alloactivated splenocytes did not. Furthermore, while normal splenocytes caused antibody‐dependent cell‐mediated cytotoxicity (ADCC) of xenogeneic endothelial cells, xenoactivated splenocytes caused significantly greater endothelial cytotoxicity by antibody‐independent mechanisms. Both normal and xenoactivated splenocytes were significantly less cytotoxic if adherent cells, consisting predominantly of monocytes and macrophages, were first removed. In vivo recipient macrophage depletion, using liposome‐encapsulated dichloromethylene diphosphonate, did not influence DXR and this may indicate that nonphagocytic circulating monocytes may be more important in DXR. However, adoptive transfer of splenocytes from a macrophage depleted, xenoactivated donor did not accelerate xenograft rejection, while splenocytes from a nondepleted xenoactivated donor did, thereby supporting the importance of monocytes and macrophages in this I phase of xenograft rejection.
Isolation of porcine pancreatic islets for xenotransplantation studies: Effects of low collagenase digestion temperatures
Authors: Sun YiSu , Zhou Daobiao , Vacek Ivan , Sun Anthony M.
Published: November 2008

Abstract: Islets of Langerhans were isolated from porcine pancreata by a modification of our previously described method. The modification involved the use of a low temperature of collagenase digestion (30°C) during the process of islet isolation. The resulting islets were then evaluated in vitro and in vivo and compared to islets isolated at the regular 37°C temperature.

The islets produced at the low temperature were more compact compared to the control islets. In the dextran density gradient these islets were deposited at the interface of the 1.060 and 1.068 g/ml density bands as compared to 1.050 and 1.060 g/ml for the control islets. In addition, the experimental islets contained a higher proportion of compact, unfragmented islets (68%) compared to the regular islets (55%), and their uptake of the dithizone stain was considerably slower than with the control islets. All ten batches of freshly isolated microencapsulated islets produced at both temperatures responded to the glucose stimulation. After 4 weeks of in vitro culture the islets of both groups microencapsulated in alginate‐polylysine‐alginate (APA) microcapsules still retained glucose responsiveness, with the experimental islets demonstrating significantly higher responsiveness to the high glucose (16.7 mM) and 0.1 mM IB MX stimulation. The morphology of unencapsulated islets in the experimental group following 4 weeks of in vitro culture indicates much firmer islet structure compared to the control islets. In addition, the unencapsulated experimental islets following the 4 week culture were still found to have secreted insulin when exposed to glucose. In transplantation studies both the experimental and the control islets normalized diabetic hyperglycemia in diabetic mice in a comparable fashion. In general, the low temperature digestion results in superior islets in terms of their morphology, viability, and physiological function.
Xenogeneic bone marrow transplantation: II. Porcine‐specific growth factors enhance porcine bone marrow engraftment in an in vitro primate microenvironment
Authors: Giovino Maria A. , Hawley Robert J. , Dickerson Matt W. , Glaser Roseann , Meshulam Deborah H. , Ardtsini Robert , Rosa Margaret D. , Monroy Rod L.
Published: November 2008

Abstract: Establishment of mixed bone marrow chimerism in pig‐to‐primate transplantation, as a means of inducing specific immune tolerance, will require that both immune and nonimmune barriers be overcome. As a preliminary step in evaluating nonimmune barriers in this system, we have developed an in vitro model of engraftment in which long‐term culture of porcine bone marrow‐derived hematopoietic cells is supported on preformed primate bone marrow stromal layers. In the absence of cytokine supplementation, primate stromal cells were unable to support long‐term porcine hematopoiesis in these cultures. Supplementation with porcine Steel Factor was required for long‐term maintenance of hematopoietic progenitor cell content and total hematopoietic activity. Addition of porcine IL‐3, in combination with porcine Steel Factor, increased long‐term progenitor cell content and hematopoietic activity on primate stroma to levels comparable to that obtained in cultures on porcine stroma. The combination of porcine GM‐CSF and Steel Factor increased progenitor cell content and hematopoietic activity early in the cultures, but had little effect in long‐term cultures. The Steel Factor and IL‐3 combination was species‐specific in its action in these cultures, as the corresponding human cytokines were unable to effectively support long‐term porcine hematopoiesis. Likewise, the combination of porcine cytokines had only minimal effects on long‐term bone marrow culture of primate CD34+ cells I on primate stroma.
A study of three candidate genes for pre‐eclampsia in a Sinhalese population from Sri Lanka
Authors: Dissanayake Vajira H. W. , Giles Victoria , Jayasekara Rohan W. , Seneviratne Harshalal R. , Kalsheker Noor , Pipkin Fiona Broughton , Morgan Linda
Published: November 2008

Aim:  The aim of these investigations was to study three candidate genes for pre‐eclampsia – epidermal growth factor (EGF), transforming growth factor alpha, and angiotensinogen – in pregnant Sinhalese women from Sri Lanka, the first such study undertaken in this ethnic group. Reproducibility of results of genetic association studies of candidate genes for pre‐eclampsia has not been consistent across populations. One of the factors that may contribute to such inconsistencies is genetic stratification due to population admixture. We therefore compared the allele frequencies of these candidate genes in healthy Sri Lankan subjects from three ethnic groups – Sinhalese, Sri Lankan Tamils and Moors – and in white Western Europeans.

Methods:  Allele frequencies were established in 80 subjects from each of four populations (Sinhalese, Sri Lankan Tamils, and Moors in Sri Lanka and white Western Europeans in the UK). A further 175 Sinhalese women with pre‐eclampsia and 171 normotensive Sinhalese controls were genotyped at eight single nucleotide polymorphisms in the candidate genes.

Results and Discussion:  In all genes haplotype and allele frequencies were comparable within the three Sri Lankan populations, but differed significantly from those in the white Western European population. Consequently cryptic population stratification is unlikely to have significant effects on allele or haplotype frequencies of the genes examined in this case–control study of Sinhalese women which showed a marginal association for EGF haplotypes and genotypes with pre‐eclampsia (P = 0.031). This association requires replication in other populations.
Maternal and fetal serum transformed alpha‐fetoprotein levels in normal pregnancy
Authors: González‐Bugatto Fernando , Foncubierta Esther , Ángeles Bailén María de los , Illanes Sebastián , Hervías‐Vivancos Blas , Bartha José L.
Published: November 2008

Aim:  To evaluate transformed alpha‐fetoprotein (t‐AFP) (a new molecular conformation of alpha‐fetoprotein) levels in maternal serum and fetal serum in normal pregnancy.

Methods:  Prospective longitudinal study. Fifty pregnant women were studied in two groups: 25 were evaluated in each trimester of pregnancy and near term (12, 20, 32 and 36 weeks) and the other 25 were evaluated at the time of planned cesarean section at term. In the first group, maternal serum t‐AFP was measured and in the second group, maternal and fetal serum t‐AFP were analyzed.

Results:  Maternal serum t‐AFP levels (medians) were 14.73 ng/mL in the first trimester, 28.29 ng/mL in the second trimester, 30.45 ng/mL in the early third trimester and 8.06 ng/mL in late pregnancy. t‐AFP levels were significantly higher in maternal than in fetal serum (P < 0.001). There were no significant correlations between AFP and t‐AFP levels in maternal versus fetal serum.

Conclusions:  t‐AFP increases during pregnancy until the early third trimester and then falls before delivery. t‐AFP levels are higher in maternal than in fetal serum which suggests that native AFP is transformed to t‐AFP either in the mother or in the placenta.
The effect of dietary protein intake on coronary heart disease risk
Authors: Papakonstantinou E. , Zampelas A.
Published: November 2008

There is cumulative evidence that there is a relationship between diet and coronary heart disease (CHD) risk. The majority of studies have focused on the role of carbohydrate and fat on CHD risk; yet the role of protein has, until recently, been neglected. The effects of dietary protein on modulating CHD risk factors are still unclear. The aim of this review is to summarise available data from epidemiological and randomised control trials on the effects of animal and vegetable protein intake on markers of CHD risk; that is, obesity, hyperlipidaemia, glucose intolerance, diabetes and hypertension. Regarding the management of body weight, reviewed data suggest that there is no convincing evidence that higher protein (HP) diets enhance weight loss compared with low protein (LP) diets, but there is some evidence that HP diets may lead to better weight maintenance. Moreover, HP diets seem to have a more favourable effect on triglyceride levels than LP diets, but on the other hand, there is no convincing evidence that there is a significant beneficial effect on total and low‐density lipoprotein cholesterol levels. In the literature, there is also a concern that higher protein intakes may increase blood glucose levels and may aggravate glycaemic control. Currently, data suggest that HP diets may have a favourable effect on postprandial glucose and glycated haemoglobin levels, whereas there is no convincing evidence on their effects on fasting glucose and insulin levels. There has also been concern that higher protein intakes may increase arterial blood pressure because of higher levels of sodium in protein‐rich foods. The data we reviewed suggest that HP diets may be more beneficial in ameliorating hypertension than LP diets. Finally, the effect of a higher protein intake on kidney function has been debated.
Mycoprotein and health
Authors: Denny A. , Aisbitt B. , Lunn J.
Published: November 2008

Mycoprotein is a high protein, high fibre, low fat food ingredient derived from fermentation of the filamentous fungus Fusarium venenatum. Interest in the putative role of mycoprotein in lowering blood cholesterol concentrations, reducing energy intakes and controlling blood sugar levels has generated a small number of human studies investigating the effects of mycoprotein on cholesterol reduction, satiety and insulinaemia/glycaemia.

In today's ‘obesogenic’ environment, in which there is an abundance of foods high in fat and/or sugar available to consumers, there is growing interest in foods that are both nutritious and satiating, but that are of low‐energy density, and are low in saturates, salt and sugar. Mycoprotein has a favourable fatty acid profile (being relatively low in saturates), a fibre content that is comparable with other vegetarian protein sources, and a naturally low sodium content. Mycoprotein is a good source of zinc and selenium but the levels of iron and vitamin B12 in mycoprotein are low in comparison to red meat.

A small number of studies investigating the cholesterol‐lowering effects of mycoprotein have been carried out among normo‐ and hypercholesterolaemic adults. The published studies to date have a number of limitations (including small sample sizes and short study durations), but overall the studies report statistically significant reductions in total cholesterol amongst hypercholesterolaemic subjects (in the order of 4–14%). These results look promising in terms of the ability of mycoprotein to contribute modest but meaningful effects on blood cholesterol concentrations, as part of a varied and balanced diet. However, the exact amount of mycoprotein that would need to be consumed in free‐living populations to have meaningful effects on cholesterol is a candidate for further confirmatory research.

A number of studies have investigated the effects of mycoprotein in comparison with other protein sources on satiety. Several studies suggest that the effects of mycoprotein on satiety are greater than an equivalent amount of chicken but it is unclear what mechanism underlies this. The studies conducted so far are relatively small, and carried out under controlled conditions, so it is difficult to extrapolate the results to larger free‐living populations.

The promotion of mycoprotein could potentially be useful, alongside other strategies, in the management of obesity and type 2 diabetes, as it appears to show beneficial effects on glycaemia and insulinaemia in the small number of studies where this has been investigated. More research is needed to better understand the mechanism of action whereby mycoprotein influences glycaemia and insulinaemia, and whether there is any dose‐dependent effect.

This paper reviews the published evidence for mycoprotein and the topics above, draws interim conclusions about the role of mycoprotein in human health and identifies areas for future research.
Perspectives for metabolomics in human nutrition: an overview
Authors: Koulman A. , Volmer D. A.
Published: November 2008

Metabolomics describes the measurement of the full complement of the products of metabolism in a single biological sample and correlating these metabolomic profiles with known physiological or pathological states. The metabolome offers the possibility of finding unique fingerprints responsible for different phenotypes. Analytical techniques such as nuclear magnetic resonance or mass spectrometry measure thousands of compounds within the metabolome simultaneously, and appropriate data mining and database tools allow the finding of significant correlations among the measured metabolomes. The first direct outcome of nutritional metabolomics will be the discovery of biomarkers, which can reveal changes in health and disease, but also indicate short‐term and long‐term dietary intake. The concerted actions of nutrigenomics and metabolomics will play a crucial role in understanding how specific interactions of single nucleotide polymorphisms influence a person's response to a diet. Finally, systems biology approaches to human nutrition combine transcriptomics, proteomics and metabolomics, with the aim of understanding how diets interact within the human being.
Real Wage Cyclicality in Italy
Authors: Peng Fei , Siebert W. Stanley
Published: November 2008

This paper analyses the cyclical behaviour of male real wages in Italy, distinguishing between North and Centre‐South, using the European Community Household Panel 1994–2001. We separate job stayers (remaining in the same job), from within‐ and between‐company job movers. Stayers are the large majority. We find stayers in the North to have high procyclicality of real wages, higher in fact than the USA and the UK. Procyclicality is highest in small firms and the private sector, as expected. In contrast, we find that real wages in the Centre‐South exhibit little cyclicality, responding rather to Northern than to local unemployment conditions.
Legislative Pay and Legislator Quality
Authors: Mixon Jr Franklin G. , Salter Sean P.
Published: November 2008

This study extends previous work by Mixon and Wilkinson (1999, Public Finance Review 27: 418–433) and Palia (2000, RAND Journal of Economics 31: 165–179) suggesting that formal human capital attainment is lower in political and managerial roles wherein expected compensation is less. The present study constructs a cumulative probability distribution function for years of education attained for both branches of the 109th US Congress and finds that the distribution of years of education attainment in the US Senate second‐order dominates that in the US House due to differences in the expected compensation levels favoring the former congressional branch. The stochastic dominance test results are supported by findings of significant differences in education attainment (favoring senators) at different quantiles of the joint education attainment distribution. Finally, goodness‐of‐fit tests also indicate that the distribution of education attainment quality in the US Senate, as measured by various academic institution quality indicators, is significantly different from that in the US House, again owing to the differences in expected compensation.
The Early‐career Gender Wage Gap among University Graduates in the Finnish Private Sector
Authors: Napari Sami
Published: November 2008

In the Finnish private sector, the gender wage gap increases significantly during the first 10 years after labour market entry, accounting for most of the lifetime increase in the gender wage differentials. This paper investigates the reasons for this gender difference in early‐career wage development. By focusing on university graduates the paper considers several explanations based on the human capital theory, job mobility, and labour market segregation. The results suggest that only about 20–26 per cent of the average early‐career gender wage gap is explained by gender differences in experience, the field of education, employer characteristics, and mobility. A substantial unexplained gap thus remains. Of the investigated factors gender differences in the field of education and work experience matter most.
Transcranial magnetic stimulation improves naming in Alzheimer disease patients at different stages of cognitive decline
Authors: Cotelli M. , Manenti R. , Cappa S. F. , Zanetti O. , Miniussi C.
Published: November 2008

Objective:  Word‐finding difficulty (anomia) is commonly observed in Alzheimer’s dementia (AD). The aim of this study was to assess the effect of repetitive transcranial magnetic stimulation (rTMS) applied to the dorso‐lateral prefrontal cortex (dlPFC) on picture naming in 24 probable AD patients with different degrees of cognitive decline.

Methods:  High‐frequency rTMS was applied to the left and right dlPFC during object and action naming in AD patients. A sham stimulation was used as a control condition.

Results:  Whilst, as previously reported, stimulation to both the left and the right dlPFC improved action, but not object naming in the mild AD group; an improved naming accuracy for both classes of stimuli was found in the moderate to severe group.

Conclusions:  Repetitive transcranial magnetic stimulation applied to the dlPFC improves naming performance also in the advanced stages of AD. Moreover, in the severe group the effect is not specific for action naming, as in the case of the mild AD group. These findings suggest that rTMS can affect the intrinsic ability of the brain to restore or compensate for damaged function and may represent an useful new tool for cognitive rehabilitation.
The long‐term functional status in patients with Guillain‐Barré syndrome
Authors: Rudolph T. , Larsen J. P. , Farbu E.
Published: November 2008

Background and purpose:  The purpose of this study was to analyse the long‐term impact of Guillain‐Barré syndrome (GBS) on quality of life, and the relationship between clinical variables at disease onset and symptoms at follow‐up to general health status.

Methods:  Forty‐two GBS patients were examined at median 6 years after disease onset and were compared with 50 healthy controls. The fatigue severity scale (FSS), visual analogue scale (VAS) for pain, disability rating index (DRI) and medical outcome study 36‐item short‐form health status scale (SF‐36) were applied. Variables at onset and symptoms at follow‐up were correlated with outcome measurements in GBS.

Results:  VAS [2.9 (SD 3.3) vs. 1.5 (SD 1.9); P = 0.01] and DRI [2.5 (SD 2.1) vs. 1.0 (SD 1.5); P < 0.001] were significantly higher in patients with GBS, compared with healthy controls. Decreased physical functioning and general health were found on SF‐36. Differences between GBS patients with shorter (<6 years) and longer (≥6 years) follow‐up after onset were not found.

Conclusions:  Relatively independent from various variables at onset, patients with GBS seem to have a reduced quality of life and functioning, and the distress seems to have become persistent after the first few years with improvement following the acute disease.
Body temperature and response to thrombolytic therapy in acute ischaemic stroke
Authors: Millán M. , Grau L. , Castellanos M. , Rodríguez‐Yáñez M. , Arenillas J. F. , Nombela F. , Pérez de la Ossa N. , López‐Manzanares L. , Serena J. , Castillo J. , Dávalos A.
Published: November 2008

Objective:  To determine the relationship between body temperature (BT), arterial recanalization, functional outcome, and hemorrhagic transformation (HT) of cerebral infarction in patients treated with i.v. tissue plasminogen activator (tPA).

Methods:  We studied 254 patients treated with tPA within 3 h from stroke onset. National Institute of Health Stroke Scale score, BT, and transcranial Doppler ultrasound (n = 99) on admission and at 24 h were recorded. Hypodensity volume and HT were evaluated on CT at 24–36 h. Poor outcome (Rankin Scale > 2) was evaluated at 3 months.

Results:  Arterial recanalization at 24 h was found in 70.7% of patients, HT in 24.8% (symptomatic in 4.7%) and poor outcome in 44.1%. Baseline BT was not associated with greater stroke severity at admission or at 24 h, HT or poor outcome. However, BT at 24 h correlated to stroke severity (P < 0.001) and hypodensity volume (P < 0.001) at 24 h, and was higher in patients who did not recanalize (P = 0.001), had symptomatic HT (P = 0.063) and poor outcome (P < 0.001). The adjusted odds ratio of poor outcome for patients with BT at 24 h ≥ 37°C was 2.56 (1.19–5.50, P = 0.016).

Conclusion:  Body temperature ≥37°C at 24 h, but not at baseline, is associated with a lack of recanalization, greater hypodensity volume and worse outcome in stroke patients treated with tPA.
Subcutaneous histamine versus botulinum toxin type A in migraine prophylaxis: a randomized, double‐blind study
Authors: Millán‐Guerrero R. O. , Isais‐Millán S. , Barreto‐Vizcaíno S. , Rivera‐Castaño L. , Rios‐Madariaga C.
Published: November 2008

Objectives: To compare the efficacy and tolerability of the subcutaneous administration of histamine and botulinum toxin type A (BoNTA) in migraine prophylaxis.

Background: Histamine has a selective affinity for H3 receptors and it may specifically inhibit the neurogenic edema response involved in migraine pathophysiology.

Methods: One hundred patients with migraine were selected in a 12‐week double‐blind controlled clinical trial to evaluate the efficacy of subcutaneous administration of histamine (1–10 ng twice a week)n = 50, compared with administration of 50 U of BoNTA (one injection cycle)n = 50.

Results: The data collected during the 4th week of treatment revealed a significant decrease in all parameters studied, in histamine and BoNTA (P < 0.001). After 4 weeks of treatment, but one injection cycle of 50 U BoNTA had only a 40‐day period of efficacy.

Conclusions: This randomized study demonstrated that both histamine and BoNTA are similarly effective and well tolerated in reducing or eliminating headache in migraine prophylaxis. Low doses of histamine applied subcutaneously may represent a novel and effective therapeutic alternative in migraine patients and lay the clinical and pharmacological groundwork for the use of H3 agonist in migraine prophylaxis.
R632W mutation in PLA2G6 segregates with dystonia‐parkinsonism in a consanguineous Iranian family
Authors: Sina F. , Shojaee S. , Elahi E. , Paisán‐Ruiz C.
Published: November 2008

Background:  PLA2G6 mutations are known to be responsible for infantile neuroaxonal dystrophy (INAD) and neurodegeneration with brain iron accumulation (NBIA). In addition, novel mutations in PLA2G6 have recently been associated with dystonia‐parkinsonism in two unrelated consanguineous families.

Methods:  Direct sequencing analysis of the PLA2G6 gene.

Results:  Here, we report the segregation of R632W with disease in an Iranian consanguineous dystonia‐parkinsonism pedigree. The identical mutation was previously observed in a patient affected with NBIA.

Conclusion:  We conclude that different and even identical PLA2G6 mutations may cause neurodegenerative diseases with heterogeneous clinical manifestations, including INAD, NBIA and dystonia‐parkinsonism.
Degenerative cervical radiculopathy: diagnosis and conservative treatment. A review
Authors: Kuijper B. , Tans J. Th. J. , Schimsheimer R. J. , Van Der Kallen B. F. W. , Beelen A. , Nollet F. , De Visser M.
Published: November 2008

Degenerative cervical radiculopathy: clinical diagnosis and conservative treatment. A review. To provide a state‐of‐the‐art assessment of diagnosis and non‐surgical treatment of degenerative cervical radiculopathy a literature search for studies on epidemiology, diagnosis including electrophysiological examination and imaging studies, and different types of conservative treatment was undertaken. The most common causes of cervical root compression are spondylarthrosis and disc herniation. Diagnosis is made mainly on clinical grounds, although there are no well‐defined criteria. Provocative tests like the foraminal compression test are widely used but not properly evaluated. The clinical diagnosis of degenerative cervical radiculopathy can be confirmed by magnetic resonance imaging. The role of electromyography is mainly to rule out other conditions. Cervical radiculopathy is initially treated conservatively, although no treatment modality has been evaluated in a randomized controlled trial. Degenerative cervical radiculopathy: diagnosis and conservative treatment. A review.