What is already known about this topic? An increasing number of studies have projected temperature-related mortality, but few consider the change of population’s adaptability to future temperature and mortality burden from cold and heat effects. What is added by this report? This study offers a comprehensive characterization of human adaptability and excess mortality burden of temperature across various regions of China. What are the implications for public health practice? The temperature-related excess mortality was projected to increase in the 2050s and decrease in the 2080s. Heat adaptability was projected to increase in the future, but along with the rising temperatures, the heat-related excess mortality continuously rose, except for the low-speed rising scenario. Although the excess mortality of cold was projected to decrease in the nearer future, it might not keep declining in the long run, due to the decreasing cold-adaptability, which deserves more attention.

Bovine Spongiform Encephalopathy (BSE) is a new fatal degenerative disease of the Central Nervous System, belonging to the unconventional slow virus diseases. These diseases are characterised by the formation and accumulation of the amyloid protein PrP in the brain of affected individuals. Thus, the development of antisera which immunostain PrP can be useful for the diagnosis of BSE. We have screened a number of polyclonal antisera prepared against the proteinase K resistant portion of PrP (PrP27–30) purified from brains of 263K scrapie-infected hamster, and ME7 scrapie-infected mouse. Moreover, we prepared polyclonal antibodies against peptides corresponding to 4 different regions of the bovine PrP purified from brainstems and cortices of five clinically suspected cases of BSE. Antisera showed that the band pattern of bovine PrP27–30 (PrPBSE) differs from that of PrP27–30 purified from brains of hamsters with experimental scrapie (PrP263K) and from brains of patients dying of CJD (PrPCJD). The amount of PrPBSE in affected brain was at least 10-fold less than that found in CJD brains and 10000-fold less than in scrapie-infected hamsters.

It has been recently shown that NGF is not only involved in the survival and development of sympathetic and neural crest-derived sensory neurons, but also in some mechanisms of the immune system. For this reason, we studied the content of NGF in CSF samples from patients with diseases in which neuroimmonological mechanisms seem to be involved (multiple sclerosis, amyotrophic lateral sclerosis, Alzheimer disease, chronic relapsing polyradiculoneuritis, Guillain-Barré syndrome, and tumors of the nervous system), as well as from a number of normal control subjects. We setup an ELISA aimed at the beta subunit of NGF, obtaining good validation tests and a detection limit of 28 pg βNGF per ml. None of the samples was found to contain detectable levels of NGF and, when a concentration method for sample enrichment was used, only one patient was NGF-positive. This suggests that NGF is probably not involved in the neuroimmunological mechanisms underlying some inflammatory and degenerative diseases of the nervous system.

The gene (NGFB) encoding the beta subunit of mature human nerve growth factor (hNGFB) was subcloned into the pJLA503 expression vector under the control of bacteriophage promoters PR and PL, and expressed in Escherichia coli. The recombinant protein represented approximately 3% of the total cellular protein. Biologically active hNGFB was solubilized (0.2% total NGFB) and purified by cation-exchange chromatography and it yielded two bands on polyacrylamide-gel electrophoresis under nonreducing conditions, corresponding to the monomeric (14 kDa) and homodimeric (26.5 kDa) forms of the molecule. Both hNGFB forms were immunopositive on Western blots with rabbit anti-NGFB antibodies; however, following additional purification, only the species corresponding to the hNGFB homodimer was biologically active on cultured chicken dorsal root ganglion neurons. These results demonstrate the feasibility of synthesizing the biologically active form of hNGFB in E. coli.

The cDNA for human ciliary neuronotrophic factor (CNTF) has been cloned into an expression vector under the control of the T7 promoter. The BL21 strain of E. coli was transformed with this vector. Human CNTF accounted for about 30% of the total bacterial protein after induction with isopropyl-B-D-thiogalactopyranoside. This human CNTF was purified to homogeneity from inclusion bodies by a combination of ion exchange chromatography and reverse-phase high performance liquid chromatography. The amino-terminal amino acid sequence of the purified protein was identical to the deduced amino acid sequence; however, the methionyl residue has been removed. On SDS-PAGE gels, human CNTF displayed a molecular weight of about 24 kDa, in accord with its deduced molecular mass; a pI of 5.8 indicates the acidic nature of the molecule. A proposed structure for human CNTF includes major alpha helical regions. The ED50 of purified human CNTF was approximately 30 pM, using cultured embryonic day 10 chicken dorsal root ganglion neurons; no activity was observed with neurons from embryonic day 8 ganglia. Polyclonal antibodies prepared against both a synthetic peptide of CNTF and the entire human CNTF protein recognized a single 24 kDa band on Western blots, corresponding to human CNTF. However, only the antibodies against intact CNTF blocked its biological activity. This represents the first molecular expression and purification of human CNTF.

Two polyclonal antibodies were raised by immunizing rabbits with two non carrier-linked synthetic peptides whose amino acid sequences corresponded to codons 89-107 (peptide P1) and 219-233 (peptide P2) of the translated cDNA sequence of murine PrP protein. These free peptides, whose structural characteristics in solution were studied by circular dichroism, elicited a reasonable immunologic response in animals. Both antibodies still recognized the corresponding immunogens after affinity chromatography purification. However, only antibodies raised to the former sequence reacted by immunoblot with a purified preparation of murine scrapie amyloid protein. These findings are discussed together with their correlation to peptide structure and the effectiveness of this simplified immunization procedure.

1. The plasma decay, tissue uptake and biotransformation of radiolabelled phosphatidylserine (PS) liposomes have been investigated in rats following bolus i.v. injection (2 mg kg-1). 2. PS plasma concentration showed a biexponential decay with half-lives of 0.85 and 40 min. The following interpretation of the biphasic decay is proposed: (1) The rapid initial decline is due to the irreversible uptake of PS liposomes by the mononuclear phagocyte system, as demonstrated by the almost exclusive accumulation of PS in liver and spleen. (2) The slow decay phase reflects the elimination of that fraction of PS that has been incorporated into high density plasma lipoproteins (HDL). A kinetic model has been developed to describe these phenomena and a good agreement has been observed between experimental data and theoretical values. 3. Evidence has been obtained that a large fraction of PS is hydrolyzed at the injection site, probably by phospholipase A2 and other hydrolytic enzymes released by platelets. Hydrolysis at the injection site has also been observed following intraperitoneal and intramuscular injections. 4. As shown by the comparative analysis of the biotransformation products found in tissues after administration of either [3H]-glycerol-PS or [14C]-serine-PS, parenterally administered PS follows two distinct metabolic pathways: (1) decarboxylation to phosphatidylethanolamine and (2) extensive hydrolytic degradation with release of the individual components of the molecule. These pathways probably reflect the two main mechanisms of PS uptake, incorporation into the plasma membrane and internalization by endocytosis, respectively.

The effects of vinblastine (VNB) and nerve growth factor (NGF) administrations were assessed on sympathetic nerve terminals by measuring the noradrenaline (NA) content in the heart, spleen and kidneys of developing animals. Six-day-old rats, treated with 0.15 mg/kg VNB on postnatal day 3 (P3) showed a dramatic decrease of NA content in all these organs. This reduction was prevented by daily administrations of NGF on P3, P4 and P5. The effectiveness of NGF in inhibiting the VNB-induced sympathectomy was related to the dose administered and to the time interval between the VNB administration and the first NGF injection given on P3. Dose-response curves to NGF (ranging from 0.01 to 0.5 mg/kg) were obtained in both heart and spleen of VNB-treated animals. Thus, this experimental paradigm provides a quantitative assessment of the NGF effects in vivo. Furthermore, the above-mentioned experimental model was utilized to evaluate the capability of GM1 to modulate NGF activity in vivo. The systemic administration of GM1 (30 mg/kg) on P3, P4 and P5, was able to potentiate the NGF activity in preventing the VNB-induced sympathectomy. This GM1 effect was more evident in the heart and may be, at least in part, attributed to increased NGF prevention of neuronal cell death due to VNB. These results suggest an in vivo interaction between exogenous GM1 and NGF and are consistent with the view that neuronal cell repair related to in vivo administration of this ganglioside may rely on its capability to modulate the activity of endogenously occuring neuronotrophic factors.

The concentration of phospholipids and proteins was determined in 23 inflammatory synovial fluids obtained from human knee joints. The synovial fluid to plasma phospholipid ratio (0.48 and 0.37 at high and low inflammatory state) was lower than the value found for the total protein content (0.68 and 0.53, respectively) indicating that phospholipids were more discriminated than proteins in their transfer from plasma to the synovial space. Constant amounts of phosphatidylinositol were found in all synovial fluids, whereas trace amounts of lysophosphatidylethanolamine and phosphatidylserine were more frequent in the active inflammatory state. A decrease in the relative amounts of phosphatidylcholine and phosphatidylinositol with respect to plasma suggested the possibility of phospholipid hydrolysis in the synovial compartment. In agreement, determinations of phospholipase activity disclosed the presence of a phospholipase A2 in the fluid phase of synovial effusions. Phospholipid derivatives formed in the synovial space may thus contribute to the amplification of the inflammatory response.

In rat peritoneal mast cells tetradecanoylphorbolacetate (TPA) induced a non cytotoxic histamine release in the absence of extracellular calcium. The addition of calcium prevented the TPA effect but micromolar concentrations of lysophosphatidylserine (lysoPS) converted the calcium-induced inhibition into a stimulation. Other lysophospholipids were inactive. In agreement with a mutual influence between lysoPS and TPA, minimal TPA concentrations enhanced the calcium-dependent histamine release induced by lysoPS in the presence of nerve-growth factor. It is proposed that the calcium-dependent pathway promoted by lysoPS and the activation of protein kinase C by TPA act synergically to induce histamine release from mast cells.

The lysophosphatidylserine-induced activation of mast cells has been studied in preparations obtained from different rodents. In mouse and gerbil peritoneal mast cells lysophosphatidylserine behaves as an agonist, inducing noncytotoxic histamine release at 0.2–8 μM. In rat peritoneal and pleural mast cells lysophosphatidylserine is ineffective, but the histamine-releasing activity becomes manifest upon the addition of suboptimal concentrations of other mast cell activators. The common structure-activity relationship shows the link between these effects of lysophosphatidylserine but the calcium requirement indicates differences in the mechanism of action. Histamine release in mouse mast cells is independent of external calcium. Thus, lysophosphatidylserine induces mobilization of endogenous calcium stores in these cells. By contrast, histamine release in gerbil and rat mast cells is dependent on the addition of external calcium indicating that the phospholipid promotes calcium influx. While in gerbil mast cells calcium influx is promoted by lysophosphatidylserine alone, in rat it requires the combined action of the phospholipid and other mast cell agonists. Differently from lysophosphatidylserine, compound 48/80 elicits histamine release in rat and gerbil mast cells. Mouse mast cells are unaffected. Thus, gerbil mast cells are the only preparation in which the action of these two agonists can be observed simultaneously.

Lysophosphatidylserine is a specific inducer of histamine release in isolated mast cells. To determine whether a similar effect is manifestin vivo, the phospholipid was injected (1–5 mg/kg i.v.) into mice and rats. A dosedependent rise in blood histamine was observed in both animals. The several-fold increase in blood histamine occurred in the first minutes and was followed by a slower decline toward normal values. A second dose of lysophosphatidylserine was without effect. Systemic manifestations (depression, hypothermia, hypotension) were associated with the increased blood histamine level. When the tissue histamine stores accessible to lysophosphatidylserine were previously decreased by repeated phospholipid injections, no systemic symptoms occurred. Mobilization of carbohydrate reserves was also manifest during the action of lysophosphatidylserine. Prior treatment with compound 48/80 induced sustained refractoriness to lysophosphatidylserine. Structure-activity relationship demonstrated that the property to induce histamine release was linked to the structure of serine head group. Thus, other natural phospholipids or lysophospholipids were inactive. It is concluded that in analogy with the effect seenin vitro lysophosphatidylserine producesin vivo release of mast cell histamine.

FEBS LettersVolume 138, Issue 2 p. 190-192 Full-length articleFree Access Interaction between nerve growth factor and lysophosphatidylserine on rat peritoneal mast cells A. Bruni, A. Bruni Institute of Pharmacology, University of Padova, Largo E. Meneghetti 2, 35100 Padova ItalySearch for more papers by this authorE. Bigon, E. Bigon Institute of Pharmacology, University of Padova, Largo E. Meneghetti 2, 35100 Padova ItalySearch for more papers by this authorE. Boarato, E. Boarato Fidia Research Laboratories, Department of Biochemistry, Via Ponte della Fabbrica 3/A, 35031 Abano Terme, ItalySearch for more papers by this authorL. Mietto, L. Mietto Fidia Research Laboratories, Department of Biochemistry, Via Ponte della Fabbrica 3/A, 35031 Abano Terme, ItalySearch for more papers by this authorA. Leon, A. Leon Fidia Research Laboratories, Department of Biochemistry, Via Ponte della Fabbrica 3/A, 35031 Abano Terme, ItalySearch for more papers by this authorG. Toffano, G. Toffano Fidia Research Laboratories, Department of Biochemistry, Via Ponte della Fabbrica 3/A, 35031 Abano Terme, ItalySearch for more papers by this author A. Bruni, A. Bruni Institute of Pharmacology, University of Padova, Largo E. Meneghetti 2, 35100 Padova ItalySearch for more papers by this authorE. Bigon, E. Bigon Institute of Pharmacology, University of Padova, Largo E. Meneghetti 2, 35100 Padova ItalySearch for more papers by this authorE. Boarato, E. Boarato Fidia Research Laboratories, Department of Biochemistry, Via Ponte della Fabbrica 3/A, 35031 Abano Terme, ItalySearch for more papers by this authorL. Mietto, L. Mietto Fidia Research Laboratories, Department of Biochemistry, Via Ponte della Fabbrica 3/A, 35031 Abano Terme, ItalySearch for more papers by this authorA. Leon, A. Leon Fidia Research Laboratories, Department of Biochemistry, Via Ponte della Fabbrica 3/A, 35031 Abano Terme, ItalySearch for more papers by this authorG. Toffano, G. Toffano Fidia Research Laboratories, Department of Biochemistry, Via Ponte della Fabbrica 3/A, 35031 Abano Terme, ItalySearch for more papers by this author First published: February 22, 1982 https://doi.org/10.1016/0014-5793(82)80438-9Citations: 132AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat No abstract is available for this article. References 1 E. Bigon, E. Boarato, A. Bruni, A. Leon, G. Toffano, Brit. J. Pharmacol., 67, (1979), 611– 616. 2 A. Bruni, E. Bigon, G. Cappellazzo, E. Boarato, A. Leon, G. Toffano, L. Amaducci Aging of the Brain and Dementia (1980), Raven New York 119– 122. 3 E. Bigon, A. Bruni, L. Mietto, G. Toffano, Brit. J. Pharmacol., 69, (1980), 11– 12. 4 T.W. Martin, D. Lagunoff, Nature, 279, (1979), 250– 252. 5 G.A. Smith, T.R. Hesketh, R.W. Plumb, J.C. Metcalfe, FEBS Lett., 105, (1979), 58– 62. 6 E. Bigon, A. Bruni, L. Mietto, E.M. Tarjan, G. Toffano, Brit. J. Pharmacol., 72, (1981), 108– 109P. 7 J.R. Carstairs, D.C. Edwards, F.L. Pearce, C.A. Vernon, S.J. Walter, Eur. J. Biochem., 77, (1977), 311– 317. 8 V. Bocchini, P.U. Angeletti, Proc. Natl. Acad. Sci. USA, 64, (1969), 787– 794. 9 R.A. Hogue-Angeletti, W.A. Frazier, J.W. Jacobs, H.D. Niall, R.A. Bradshaw, Biochemistry, 15, (1976), 26– 34. 10 L.A. Greene, Dev. Biol., 58, (1977), 106– 113. 11 D. Lagunoff, Biochem. Pharmacol., 21, (1972), 1889– 1896. 12 P.A. Shore, A. Burkhalter, V.H. Cohn, J. Pharmacol. Exp. Ther., 127, (1959), 182– 186. 13 S. Varon, J. Nomura, E.M. Shooter, Biochemistry, 6, (1967), 2202– 2209. 14 R.A. Bradshaw, Annu. Rev. Biochem., 47, (1978), 191– 216. 15 H. Thoenen, Y.A. Barde, Physiol. Rev., 60, (1980), 1284– 1335. 16 R. Merrell, M.W. Pulliam, L. Randono, L.F. Boyd, R.A. Bradshaw, L. Glaser, Proc. Natl. Acad. Sci. USA, 72, (1975), 4270– 4274. 17 P. Vadas, S. Wasi, H.Z. Movat, J.B. Hay, Nature, 293, (1981), 583– 585. Citing Literature Volume138, Issue2February 22, 1982Pages 190-192 ReferencesRelatedInformation

In the presence of mouse plasma, lysophosphatidylserine stimulates histamine secretion from isolated mast cells. The extensive modification of carbohydrate metabolism produced by lysophosphatidylserine in mice was largely prevented by the antihistaminic drug, pyrilamine. However, to prevent completely the change in carbohydrate metabolism induced by lysophosphatidylserine the administration of an antihistamine and an adrenoceptor antagonist was required. It is concluded that the effect of lysophosphatidylserine in mice is due to release of intracellular amines. Histamine and catecholamines are involved.