we (784 Effects of Growth, Fasting, and Trauma on the Concentrati ‘Connective Tissue Hexosamine and Water. (21203) Nowstax F, Boss axo Josep B. Fouty. From the National Institute of Arthritis ond Metabolic Diseases, National Inst "Mh, Health Service, U. §. Department of Health, Education end Welfare, Bethesda, We9 for 18 hours, reweighed, then extract’ Soxhlet for 3 hours with redbtiled The dry, eRe Hexosaininest are present in the ground substance of connective tissue as components ‘of mucopolysaccharides(!) and mucoproteins (2). Sinee the ground substance is altered in samples were ela the collagen diseases, it has been suggested contents calculated. The amount of ‘by many that changes in mucopalysaccharides wet sample of erase nce to these obverved lesions. may vary from 2-407, causing a Wide Ey ‘2, | Scblamowita ‘al.(3) have demonstrated that tion in calculated water and hensaciae S14 “nodules”, experimentall si pealyzing induced by injec- values from sample to sample. The Samara eos of trypsin and other enzymes, contain in- trations of tissue hexosamine and wale creased concentrations of hexosamine. This independent of the amount of fat: rose Jereiio been noted in nodules obtained from therefore, all expressions of concentra vetints with rheumatic fever(2) and in the water and hexosamine are presented Porium and subcutaneous connective tissue of to fat-free tissue. Water exprewed at pe patients with scleroderma(4). In ‘order to cent (wet wt—dry wt) x 100/(wet te | ea te Whe factors tauencing con. and hexosamine as mg/100 g of fat served a5 6 sents, Co seca body ss 100 gd Set tissue hexosamine levels, the effects of tissue. To determine the bexosaming 62> tent, the dry defatted samples were ee growth, fasting and various traumatic agents have been studied. - placed in 10m glass-stoppered aoe a Methods, Male Sprague-Dawley rats were Sik After adding one rl of 4 3 ERS eneneee aes fala dock ict(S) and Hoc acid, they were hydrolyzed |S veaieiermitied wo. drink water od libitem. 200°C: taken to volume Witt water aad ae tin ae cach cx. tered, Hlexosamines were deteminid bi ree eet. At he end of an experiment the ™bdifiation ofthe Elon wd erg Periment hed with ether, Tn ikose instances ter separation from the hydreliagy TE Tuech orbilal connective. tissue was ob- columas of Dowex-50(6), tained the rats were killed by exsanguination funder ether anesthesia. Subcutaneous con- nective tissue samples were obtained from an frea of the back, caudal to the scapular adi pose tissue. The skin was eut along the Tine of the back, retracted and the sample (60-200 mg) removed by scraping the loose connective tissue from the underside of the Coriuum with a. pair of wing-tipped forceps. ‘Orbital connective tissue was obtained follow {ng removal of the eyeballs by dissection from the Harderian glands! After weighing, all the ‘sation an i se secre ‘1c instance 28 probabil Festing fice (wns 800% (0% WATER (PERCENT) : Re stwere placed in an oven at TOL-ION"C yy & The Z amino supers normally encountered in eae aa i 212 and 3 ¥ he medod of Elson and Morgan wed here does (ycenieations in ipetacons couneetre king on th not distinguish betireen ther ‘of rate of racy Ses 3 eeCoxwecrive Tissue Hexosa: oor ations of ‘Results, Growth, Four groups of rats, 5, the experiment), The normal relationship be-| 13) ze, §,and 11 weeks of age, with 46 rats in each tween water and hexosamine wes thos alvened Beroup, Were used. Subcutaneous connective by the second day of fasting (compare with Bisse samples were obtained and analyzed Fig. 1). During this fast the mean. body tutes of Heabh, ig. 1). The water and hexosamine concen- Weight fell from 109 = 7 4 at the start of the Bechesd, Ma tons, high in the youngest group, de- €xPeriment to $0 3 4 g on the third day Greased with increasing age. Water and hexo. Since fasting did not affect besosamine loctto then extracted in z [gine values obtained on the same samples in connective tissue, food intake was not edistlled petroleus each animal were found to have a high ™easured in subsequent experiments The kiss faire Pabfesee of correlation (r = +082), which was Loc trauma. Following a preliminary ob- [ the water and {at cant at a probability Ievel of 0.01. servation that epinephrine injections increased Elbe effect of body weight on the concen- the hexosamine concentration at an injection Pixtions of water end hexosamine in orbital - site, an experiment was designed to deternine 7 ective tissue ‘was similarly studied by the tate of this change. In the frst experi, lvzing the data from 93 rats which had ment, 45 rats weighing 110-130 g were weed, Bred as controls in several different experi- § were used as an initial conteol group; a Riis, Conelation studies were made be- group of 20 rats was injected subcutaneously hee body weight (range was 60-235 g) and twice daily in the same site (right dorsum) Bexosamine concentration (range was 357-493 with O.1 ml of physiological saline; a group amount of fat in z us connective tissue using a wide vars. Cand hexosamine ple. The conces vine and water a: int of fat prese if concentration fe PRAMen TGS J EBe100 s ey wt), body veight and water of 20 rats vas injected snilaly wih Go ea Too eaten ih aumeeteation (range was 76.6-70.8%), and of epinephrine (123000), Five tats in eae On deggie de 4 Speesmine and water concentrations, There injected group were sacifced at 6 2h aa, © hie PRES sienitcant negative correlation betveen and 72 hours and comective tigve, aoiphe e bes Pine « : Eiusamine concentration and body weight obtained (Fig. 2). In both groups the tow, Ge — 81), and also between water concen- centrations of water’ and. hexsemine ins aN hee fpgegion and body weight (e=~—77). ‘Thus, creased, reaching highest obseencd.eahes vc nad of 4 hye increasing body sive there was a progres. 24 hours (epinephrine) and 48 hours (sane) rolyzed 13 beuts decrease in the concentrations of heyo- The increases in the hexosamine levels in re: Bie and water. The hexosamine and water sponse to saline and epinephrine were not sige 8 were tested and found to have a sig- nificantly diferent. At 24 hours, the increas " positive correlation (r= -} 69). In in Water concentration produced by epintghe instance the values for r were significant rine was greater than that produced by saline probability level of <.01. Jn subsequent studies, a response time of 48 ting. 8 reduced food intake decreases eEtoount of plasma hesosamine(S), Tt was Eilore considered of interest to assess the ‘of fasting on the concentrations of heno- Bee in normal connective tissue. Sixteen Bevere used, and sacrificed in groups of 4. BE croup was killed at the start of the ex: fEkoeot and the remaining groups following 2, and 3 days of tol fasting, ‘The rats gee drinking water available. [Easting had no Geant etfect on the cobtentrations of suming which were $81 - 20, 599 = 20, amples were «x? sith water, anc! £ - determined y= pd Morgan mete @ hydrolysates [ sume +f Berens 6, Gnd 584 2c 11 mp/100 g dry wt on a 2, and 3 days, respectively. [There was Five i nouns eon a eet went 20 8, Bt of he tt ep St with 81.6 296 at the start of | '* MME connec arash 21 Vineste=i injected with jution) eating cted with tan pour fast. Ail following the swater conte jth $5.3 = 4 | Hbitup), the injected grou e plasma bes re as ine the exossmin “tissue patio ctress the ed under et > sample Sment anol Ie ce the Ha tne oF ce teres” Hosa Pantornesie Set Dericrescy v that mucopolysaccharides, which 2¢- {Bont for approximately 50% of the total connective tissue hexosamine(8) and bebave ge hedrophilic colloids, would largely account t the parallelism between the tissue water (Gerd hesosamine concentrations lation between hexosamine and water has ficen observed in experimental myxedema(9), Ged in umen skin(4). This relationship is ted rats and in trau- is well recognized that following many Pes of Body injury the plasma hexosamine increases(3,5,10). The site of produc. ® oF source of this hexosamine has nat been feined. ‘The synthesis of hexosamine by iso. ed connective tissue cells has recently, been enstrated(11) and it therefore is possible t the increased amount of plasma. hexo- ine following trauma may be produced by Although it was not possi. to detect changes in hexosamine concen: ation in non-traumatized connective tissue Palowing ‘trypsin injections(3) or fracture, as sBperted here, the possibility thet a concurrent in plasma hexosamine(3,5) may re- is by connective tissue is 1. The concentrations of hexo- ‘and water in connective tissue decrease growth in the rat. ‘There is a high de- of correlation between the concentrations Bigater and hexosamine in normal connective "Pantothenic Acid Deficiency Induced in Human Subjects. tissue, 2. Most traumatic agents. (physical and chemical) effect local non-specine ine crease in hexosamine and water concentra. tions, The degree of change of each of these constituents varies with the agent adminis, tered. Fasting does not influence this. in tease in hexosamine in response to. local trauma, or the hexosamine concentration in untraumatized connective tissue, 3. Changes in the concentration of counective tissue hexo. samine were not detectable following systemic stress (fracture), 3. Meyer, Rand Rapport, NM. M, Sawa, 1951, wit3, 566 2. Consdea, R, Glenn, L. Biochem, J, 1983, x35, 299. 3 Schlomowitz, ST, DeGraf, A. Cy and Schubert, M., Circulation, 1950, v1, 82, 853, sa 4 Boas, NF, Van Scott, E. Ju and Bunim, J. Jy in preparation, Boat, N. F, and Peterman, A. F, Proc. Soc. EXE. Boor. 409 Mem, 1955, v82, 19.) 6 Boas, NF, J. Biel. Chem, 1958, os Geo Sobel Hy Zutrauen, H. A, and Marmorston, J, rch, Biock. and Bioph, 1953, v46, 2, 8. Boas, N.F, in preparation, 9. Ludiviz, AW. Boas. N. Fy and L. J. Soffer, Proe. Soe, Exe, Biot. 480 Mio, 1950, vis, 187° 10, West, R, snd Clnks, D. Hf, J. Cli, Toes, 1938, v17, 173. 21. Bolle A. J, Boas, N. F, and Bunion, J. J Science, 1958, v120, 348, E, and Suanier, W. My Received June 29, 1938. PSERBE i9ly ven, (21204) WriitaM B. Bea ax Roper E, Honcrs, Ome the Ae alter the classic demonstration: by (1) of the molecular antagonism of Faamide by para-amino benzoic acid, one SS (WBB) became interested in seeking relationships between individual Be Departnent Trust Company, The Nu- "edation end the National Vitamin Founda. ale Ward, Department of Internal Medicine, Sate University of owe, Tews City. complex vitamins and their chemical ana~ logues. A series of pyridine compounds was studied for vasodilating effects and therapeutic Potency against pellagra(?2). . Observations at that time suggested thst some of the com. pounds might have toxic properties, ‘This sus, bicion had ipeen expressed by Woolley(3) as & result of his work with canine ‘blacktongue Mellwain(4) suggested that pyridine-Soul,