Enhancement of hormonal activity with a monoclonal antibody specific to porcine growth hormone

Abstract

The potentiation of the biological effects of recombinant porcine growth hormone (pGH) by immunologic manipulation was investigated. A monoclonal antibody (mAb), designated PS‐7.6, was raised against pGH and repeatedly shown to enhance the responsiveness of hypophysectomized (hypox) rats to pGH. As a result, animals receiving a combination treatment of pGH and mAb PS‐7.6 together gained significantly more weight than those receiving the same doses of pGH alone. The enhancing action of the mAb was a rapid process and its effective doses ranged from 0.1 to 2 mg/injection. The ability of the antibody to augment the hormonal activity persisted beyond the 5‐day treatment period and the differences in net weight gain between treated and control animals remained significant for 28 days. Results from treatment frequency studies further suggested that pGH when complexed with mAb PS‐7.6 required fewer injections and produced a greater efficacy than being administered alone. Therefore, present findings suggest that mAb PS‐7.6 may prove useful for not only improving the efficiency of pGH, but also developing a novel formulation for sustained pGH release.     

Differential response to hepatic differentiation stimuli of amniotic epithelial cells isolated from four regions of the amniotic membrane

Human Amniotic Epithelial Cells (hAEC) isolated from term placenta are a promising source for regenerative medicine. However, it has long been debated whether the hAEC population consists of heterogeneous or homogeneous cells. In a previous study, we investigated the characteristics of hAEC isolated from four different regions of the amniotic membrane finding significant heterogeneity. The aim of this study was to evaluate the hepatic differentiation capability of hAEC isolated from these four regions. Human term placentae were collected after caesarean section and hAEC were isolated from four regions of the amniotic membrane (R1-R4, according to their relative distance from the umbilical cord) and treated in hepatic differentiation conditions for 14 days. hAEC-derived hepatocyte-like cells showed marked differences in the expression of hepatic markers: R4 showed higher levels of Albumin and Hepatocyte Nuclear Factor (HNF) 4α whereas R1 expressed higher Cytochrome P450 enzymes, both at the gene and protein level. These preliminary results suggest that hAEC isolated from R1 and R4 of the amniotic membrane are more prone to hepatic differentiation. Therefore, the use of hAEC from a specific region of the amniotic membrane should be taken into consideration as it could have an impact on the outcome of therapeutic applications.     

The "Alzheimer's disease signature": potential perspectives for novel biomarkers

Alzheimer's disease is a progressive and neurodegenerative disorder which involves multiple molecular mechanisms. Intense research during the last years has accumulated a large body of data and the search for sensitive and specific biomarkers has undergone a rapid evolution. However, the diagnosis remains problematic and the current tests do not accurately detect the process leading to neurodegeneration. Biomarkers discovery and validation are considered the key aspects to support clinical diagnosis and provide discriminatory power between different stages of the disorder. A considerable challenge is to integrate different types of data from new potent approach to reach a common interpretation and replicate the findings across studies and populations. Furthermore, long-term clinical follow-up and combined analysis of several biomarkers are among the most promising perspectives to diagnose and manage the disease. The present review will focus on the recent published data providing an updated overview of the main achievements in the genetic and biochemical research of the Alzheimer's disease. We also discuss the latest and most significant results that will help to define a specific disease signature whose validity might be clinically relevant for future AD diagnosis.     

Population genetics of coagulant factor IX: frequencies of two DNA polymorphisms in five ethnic groups.

Two frequently used restriction-enzyme polymorphisms (RFLPs) of coagulant F.IX, TaqI and XmnI, have been examined in five ethnic groups: white Americans, black Americans, East Indians, Chinese, and Malays. There is a distinct "cline" in the frequencies of both polymorphisms, from white Americans to Malays. The rarer type 2 alleles of both polymorphisms, in which middle recognition sites are present--and which in our sample reach their highest frequencies in white Americans--are marginally higher in four groups of Europeans previously reported by others. The frequencies of the rarer alleles are significantly higher in Europeans than in black Americans and East Indians, and these alleles are essentially absent in Chinese and Malays. The frequency of heterozygosity diminishes in the same order, being zero in Malays for both polymorphisms. The polymorphisms are in strong linkage disequilibrium, and in all groups the type 1 allele for TaqI is disproportionately accompanied by the type 1 allele for XmnI. The paucity of type 2 alleles and the low rate of heterozygosity in four non-European groups suggest that the polymorphisms will be of little diagnostic value south of Gibraltar and east of Suez. This prediction is confirmed by the observed haplotype frequencies in the black American and the Oriental groups.     

Characterization of DNA sequences that mediate nuclear protein binding to the regulatory region of the Pisum sativum (pea) chlorophyl a/b binding protein gene AB80: identification of a repeated heptamer motif

Two protein factors binding to the regulatory region of the pea chlorophyl a/b binding protein gene AB80 have been identified. One of these factors is found only in green tissue but not in etiolated or root tissue. The second factor (denominated ABF-2) binds to a DNA sequence element that contains a direct heptamer repeat TCTCAAA. It was found that presence of both of the repeats is essential for binding. ABF-2 is present in both green and etiolated tissue and in roots and factors analogous to ABF-2 are present in several plant species. Computer analysis showed that the TCTCAAA motif is present in the regulatory region of several plant genes.     

The malmö polymorphism of coagulation factor IX, an immunologic polymorphism due to dimorphism of residue 148 that is in linkage disequilibrium with two other F.IX polymorphisms

A mouse monoclonal antibody (MAB 9.9) to coagulation factor IX (F.IX) detects a polymorphism in the plasma of normal people. Its epitope has been narrowed down to <6 amino acids in the activation peptide of the X-linked F.IX protein. The activation peptide contains a dimorphism—Thr:Ala—at position 148 of the protein. Using synthetic oligonucleotides, we have demonstrated that (1) the F.IX which reacts with 9.9 has Thr at position 148 and (2) that which does not has Ala. Positive reactors (148^thr) are designated Malmö A, and negative reactors (148^ala) are designated Malmö B. The plasma levels of AA women are indistinguishable from those of A men, and both B men and BB women are null against MAB 9.9. The plasma level of Malmö A in AB women is approximately half that of AA women, and “lyonization” is clearly operating in the heterozygotes. The dimorphism is in strong linkage disequilibrium with two other intragenic RFLPs, TaqI and XmnI. Furthermore, intragenic crossing-over—including double crossing-over—appears to have occurred between the three sites. Seven of the eight possible haplotypes have been identified, five in men and two others in women. The immunoassay that identifies ～50% of the AB women in the pool of Malmö A females with 95% confidence identifies men unambiguously as A or B. The assay would be very useful for population-genetic studies of the Malmö epitope if the studies were limited to men.     

Eye factors and lens-forming transformations of outer cornea in Xenopus laevis larvae

Outer cornea of lensectomized Xenopus laevis tadpoles at state 50 (according to Nieuwkoop, P.D. and Faber, J., ('56) Normal Table of Xenopus laevis, Daudin, North-Holland, Amsterdam, pp. 1-243) was removed 3, 7 and 10 days after lensectomy and implanted between the outer and the inner cornea of larvae of the same species at stage 51-52. In these conditions, the implanted outer cornea remained isolated from the retinal factor of the vitreous chamber, although it received the nutritional factors normally reaching the outer cornea. Results show that lens-forming transformation process of the outer cornea is arrested, and lens-forming structures undergo regression at speed which increases with increasing precocity of the stage of lens-forming transformation undergone by the implanted cornea. These data suggest that the process of lens-forming transformation is not a single-step process, but a sequence of interactions extending over a long period of time requiring the continuous presence of the retinal factor in the vitreous chamber until complete differentiation of the lens is achieved.     

Spontaneous binding ofYersinia enterocolitica to murine leukocytes

Twelve strains ofYersinia enterocolitica were examined for their ability to bind spontaneously to murine leukocytes. Each of eight HeLa cell invasive strains exhibited nonselective binding to peritoneal leukocytes, lymph node leukocytes, and thymocytes, whereas four noninvasive strains lacked binding properties. Like the HeLa cell invasion, the binding ofY. enterocolitica to leukocytes was much less efficient for bacteria grown at 37°C than for bacteria grown at 22°C. The binding properties were not influenced by the virulence plasmid that codes for Vwa⁺ phenotype. This leukocyte binding test is proposed as a simple assay for invasive properties ofY. enterocolitica.     

Thirst and artificial heat acclimatization in man

The purpose of this study was to investigate the relationship between serum osmotic changes, water intake and water balance in four, fit young men during and after exercise in the heat, before and after artificial heat acclimatization. During exercise, before steady-state conditions were reached, voluntary water intakes generally paralleled but were not proportional to the serum osmotic pressure. In steady-state conditions, drinking was approximately proportional to the effective osmotic pressure of the serum. During the post-exercise recovery period, when serum osmolarity returned to normal levels, water intake also subsided even though there was a total body water (wt) deficit of about 3%. Body weight did not return to control levels until 62 to 86 hr following the exercise. This slow recovery could not be accounted for by a loss of water associated with glycogen utilization during exercise or by sweat electrolyte depletion. In general, the results supported Dill's hypothesis but plasma volume changes, in addition to osmotic factors,are very likely operative in the initiation and satiation of drinking under these conditions. Perhaps slower acting volume control mechanisms mediate the slow recovery of total body water.

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Zusammenfassung Untersucht wurde die Beziehung zwischen osmotischen Veränderungen im Serum, Wasseraufnahme und Wassergleichgewicht bei 4 gesunden,jungen Männern während und nach körperlicher Arbeit in der Hitze vor und nach künstlicher Hitzeakklimatisation. Die freiwillige Wasseraufnahme während der Arbeit, ehe ein steady state erreicht wurde, war im allgemeinen gleichlaufend, aber nicht proportional dem osmotischen Serumdruck. Im steady state war die Trinkmenge ungefähr proportional dem effektiven osmotischen Serumdruck. Wenn die Serumosmolarität während der Erholungsphase zur normalen Werten zurückkehrte, liess die Wasseraufnahme ebenfalls nach,obwohl das Gesamtkörperwasserdefizit ungefähr 3% betrug.Die Körpergewichte erreichten die Ausgangswerte 62–86 Std nach der Arbeit. Diese langsame Erholung war nicht bedingt durch einen Wasserverlust dur Glykogenverwertung während der Arbeit oder durch Elektrolytverluste beim Schwitzen. Die Ergebnisse stützen Dill's Hypothese, doch unter diesen Bedingungen wirken Plasmavolumen-Veränderungen zusätzlich zu osmotischen Faktoren sehr wahrscheinlich mit bei der Auslösung des Trinkens und der Sättiging mit Wasser. Vielleicht vermitteln langsamer wirkende Volumenkontrollmechanismen die langsame Wiederherstellung der Gesamtkörper-Wasservorräte.

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Resume On a examiné les relations existant chez 4 jeunes hommes sains entre les variations de la pression osmotiques du sérum d'une part,l'ingestion d'eau et l'équilibre acqueux du corps d'autre part et cela pendant et après un travail corporel à la chaleur tant avant qu'après une acclimatisation artificielle au chaud.L'ingestion volontaire d'eau évolue similairement à la pression osmotique du sérum, mais ne lui est pas proportionnel. Ce phénomène s'observe jusqu'à ce qu'un état dit stationnaire soit atteint. Dès ce moment, les quantités bues sont à peu près proportionnelles à la pression osmotique effective. Pendant la phase de récupération, la pression osmotique redevient normale et l'eau ingérée diminue bien que le déficit en eau pour le corps entier soit encore de 3% environ.Le poids du corps ne retrouve sa valeur initiale que 62 à 86 heures après la période de travail. Ce lent rétablissement n'est pas dû à des pertes d'eau par suite de l'utilisation de glycogènes pendant l'exercice ou par suite de la perte d'électrolytes par la sueur. Ces résultats viennent confirmer les hypothèses de Dill. Pourtant, dans ces conditions, les variations de volume du plasma agissent très vraisemblablement en plus des facteurs osmotiques dans le besoin de boire ou l'état de satiété. Il est possible que des mécanismes de contrôle du volume agissant plus lentement président aux phénomènes de rétablissement des réserves en eau dans son ensemble.

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Results were presented in part at the "Symposium on Nutrition and Physical Activity", The Swedish Nutrition Foundation, Tylösand, Sweden, 15–17 August,1966.