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Deitel, M, Stone, E, Kassam, HA, et al. Gynecologic-obstetric changes after loss of massive excess weight following bariatric surgery. Journal of the American College of Nutrition. 1988; 7: 147-153. IIb ; Jarow, JP, Kirkland, J, Koritnik, DR, et al. Effect of obesity and fertility status on sex steroid levels in men. Urology 1993; 42: 171-174. IIb ; Stanik, S, Dornfeld, LP, Maxwell, MH, et al. The effect of weight loss on reproductive hormones in obese men. J Clin Endocrinol Metab 1981; 53: 828-832. IIb ; Bates, W. Nutritional aspects of infertility. In: Hammond, MG, Talbert, LM, eds. Infertility. A Practical Guide for the Physician. Third Edition. Boston: Blackwell Scientific Publications; 1992: 182-195. IVa ; Sugerman, HJ, Felton III, WL, Salvant, JB, et al. Effects of surgically induced weight loss on idiopathic intracranial hypertension in morbid obesity. Neurology 1995; 45: 1655-1659. IIb ; Amaral JF, Tsiaris, W, Morgan, T, et al. Reversal of benign intracranial hypertension by surgically induced weight loss. Arch Surg. 1987; 122: 946-949. IIb ; VanItallie, TB, Lew, EA. Assessment of morbidity and mortality risk in the overweight patient. In: Wadden, TA, VanItallie, TB, eds. Treatment of the Seriously Obese Patient. New York: The Guilford Press; 1992: 3-32. IVa ; Hagen, J, Deitel, M, Khanna, RK, et al. Gastroesophageal reflux in the massively obese. Int Surg. 1987; 72: 1-3. IIb ; Egger, G. The case for using waist to hip ratio measurements in routine medical checks. The Medical Journal of Australia. 1992; 156: 280-285. IVa ; Sjostrom, CD, Hakangard, AC, Lissner, L, et al. Body compartment and subcutaneous adipose tissue distribution -- risk factor patterns in obese subjects. Obesity Research. 1995; 3: 9-22. IIa ; Folsom, AR, Kaye, SA, Sellers, TA, et al. Body fat distribution and 5-year risk of death.
FIG. 6. pCoxIV-DHFR boundto outer membrane vesicles or acidic liposomes lacks a protease-resistant DHFR moiety. A , native pCoxIV-DHFR 9 nM ; was incubated with outer membrane vesicles 20 p M ; for 20 min in 0.6 ml of 20 Tris-C1 pH M 7.0 ; , 50 mM KC1, 0.2% BSA. The fusion protein bound to vesicles 100 ; at 30 "C for 20 min in the presence of the indicated concentrations of adriamycin in 20 m Tris-C1 pH 7.0 ; , 50 m of was then re-isolated by ultracentrifugation 122, 000 g, 10 min ; and M M the vesicles were solubilized with the same buffer 0.33 ml ; containing KCl, and 0.2% BSA. 0.5% octyl-POE. Aliquots 0.05 ml ; of the solubilized fraction were incubated with the indicated amounts of trypsin micrograms ml ; a t whether the fusion protein bound to outer membranes or 0 "C for 10 min, and the digestion was then blocked by the addition acidic liposomes Fig. 6 ; was as trypsin-sensitive as the trans- of 1 m phenylmethylsulfonyl fluoride leftpanel, bound precursor ; . M pCoxIV-DHFR 50% of the amount added to the location intermediate bound to the mitochondrial surface As a control, native vesicle suspension ; was digested under the same conditions in the Eilers et al., 1988 ; . Outer membrane vesicles or liposomes CL POPG POPE presence of the same concentrations of detergent-solubilized outer membrane vesicles right panel, control precursor ; . Samples were fusion protein, analyzed by SDS-polyacrylamide gel electrophoresis and fluorograPOPE 5 20 30 were allowed to bind the re-isolated, solubilized with the non-ionic detergent octyl- phy. B, native pCoxIV-DHFR 9 nM ; was incubated with phosphoPOE, and incubated with increasing amounts of trypsin left lipid vesicles CL POPG POPE POPC 5 20 30 panels, bound precursor as a control, native fusion protein 30 "C for 20 min in 0.6 ml of 20 Tris-C1 pH 7.0 ; , 50 mM KCl, was exposed to trypsin in the presence of solubilized outer 0.2% BSA. The fusion protein bound to vesicles was then re-isolated membranes or liposomes under the same conditions right by centrifugation 15, 000 X g, 20 min ; and solubilized with the same buffer 0.33 ml ; containing0.5% octyl-POE. All other conditions were panels, control precursor ; . The bound protein was completely as in Fig. 6 except that thecontrol rightpanel ; contained solubilized digested by low concentrations of trypsin without accumula- phospholipid vesicles instead of outer membranes.

1, 25dihydroxy vitamin D3 receptor on sodium dodecyl sulfate polyacrylamide gels by renaturation and immunoblotting. Proc. Nat. Acad. Sci. USA 82: 7825, 1985. Delmas, P.D.: Treatment of postmenopausal osteoporosis. Lancet 359: 2018, 2002. DeLuca, H.F.: The vitamin D hormonal system, Pharmacological Modulation of Steroid Action'da Ed.: E. Genazzani ve di. ; , s.11, Raven Press, New York, 1980. DeLuca, H.F.: New Concepts of vitamin D function. Ann. N.Y. Acad. Sci. 660, 59, 1992. Dibble, J.B. ve M.S. Losowsky: Osteomalacia in chronic liver disease. Brit. Med. J. 285: 157, 1982. Drake, S. ve di.: Pamidronate sodium and calcitoninresistant Paget's disease. Arch. Int. Med. 49: 401, 1989. Editorial: Treatment of Paget's disease. Lancet 1: 955, 1971. Editorial: Fluoride and the treatment of osteoporosis. Lancet 1: 547, 1984. Editorial: Osteolytic Paget's disease. Lancet 1: 1255, 1986. Favus, M.J. ve di. Ed. ; : Primeron the Metabolic Bone Diseases and Disorders of Mineral Metobolism, 3. Bask , LippincottRaven, Philadelpha, 1996. Fleisch, H.: Biphosphonates in Bone Diseases. From Laboratory to the Patient. 2. Bask , Partenon, New York, 1995. Fraioli, F. ve di.: Subarachnoid injection of salmon calcitonin induces analgesia in man. Eur. J. Pharmacol. 78: 381, 1982. Haussler, M.R. ve T.A. McCain: Basic and clinical concepts related to vitamin D metabolism and action. N. Engl. J. Med. 297: 974 ve 1041, 1977. Haussler, M.R. ve P.E. Cordy: Metabolites and analogues of vitamin D. Which for what. JAMA 247: 841, 1982. Heany, R.P.: Thinking straight about calcium. N. Engl. J. Med. 328: 503 , 1993. Hillyard, C.J. ve di.: Katacalcin: a new plasma calciumlowering hormone. Lancet 1: 846, 1983. Hohmann, E.L. ve di.: Innervation of periosteum and bone by sympathetic vasoactive intestinal peptidecontaining nerve fibers. Science 232: 868, 1986. Koshy, K.T.: Vitamin D, an update. J. Pharmaceut. Sci. 71: 137, 1982. MacIntyre, T.: Calcitonin for prevention of postmenopausal bone loss. Lancet 1: 900, 1988. Malluche, H.H. ve M.C. Faugere: Renal osteodystrophy. N. Engl. J. Med. 321: 317, 1989. Matthews, E.K.: Calcium and membrane permeability. Brit. Med. Bull. 42: 391, 1986. Morel, F.: Sites of hormone action in the mammalian nephron. Am. J. Physiol. 240: F 159, 1981. Motokura, T. ve di.: Parathyroid hormonerelated protein in adult Tcell leukemialymphoma. Ann. Int. Med. 111: 484, 1989. Mundy, G.R. ve L.G. Raisz: Drugs for disorders of bone: pharmacological and clinical considerations. Drugs 8: 250, 1974. Nightingale, S.L.: Etidronate disodium approved. JAMA 258: 2487, 1987. Nilas, L. ve di.: Calcium supplementation and postmenopausal bone loss. Brit. Med. J. 289: 1103, 1984.

7. LOE, H., and SILNESS, J.: Periodontal Disease in Pregnancy: I. Prevalence and Severity, Acta Odontol Scand 21: 533-551, 1963. LINDHE, J.; HAMP, S.E.; and LOE, H.: Experimental Periodontitis in the Beagle Dog, J Periodont Res 8: 1-10, 1973. MARGALIT, D., and GEDALIA, I.: Release of Fluoride into Saliva After Topical Fluoride Application, J Dent Res 48: 93-96, 1969. AASENDEN, R.; BRUDEVOLD, F.; and RICHARDSON, B.: Clearance of Fluoride from the Mouth After Topical Treatment or the Use of a Fluoride Mouth Rinse, Arch Orat Biol 13: 625-636, 1968. LIBRUs, H.; PIETROKOVSKI, J.; ULMANSKI, M.; and GEDALIA, I.: The Effect of Fluoride on Molar Socket Healing in the Rat, Arch Oral Biol 18: 1283-1289, 1973 and flurbiprofen.

Previous studies from this laboratory have shown Morphological studies have shown that Sertoli cells and that Sertoli cell-enriched culture medium contained germ cells are closely associated in the adluminal comparttwo immunologically and structurally related proteins ment of the seminiferous epithelium and that the tight juncdesignated CMB-22 and CMB-23 with M, of 37, 000 tional complexes formed by the Sertoli cells create the bioodand 40, 000, respectively. We have now demonstrated testis barrier that isolates the processes of spermatogenesis that both CMB-22 and CMB-23 are monomeric pro- and spermiogenesis from the somatic cells and theirproducts teins with thefollowing NH, -terminal amino acid se- 1-3 ; . Thus, Sertoli cells play an importantrole in regulating quences: CMB-22, NH2-TPDPSLDVEWNEWRTKHG- germ cell development by determining the ~icroenvironment KTYNMNEERLKR; CMB-23, NH2-XAPXPDPSLDV- of the seminiferous tubules 4 ; . During the pastdecade, studEXNEXRTK. These sequences are virtually identical except that CMB-23 has three extra NH, terminus ies have shown that Sertoli cells are the major secretory amino acids of X-A-P. Comparison of these sequences component in the testis in that they synthesize and secrete with those in the Protein Identification Resource re- most, if not all, of the abundantmacromolecules required for vealed that they are unique proteins. CMB-22 and spermatogenesis behind the blood-testis barrier including CMB-23 are highly concentrated in testes and their rABP' 1, 5 ; , transferrin 6 ; , ceruloplasmin 7 ; , clusterin levels in this tissue increase with age. Studies using sulfated glycoprotein-2 ; 8-ll ; , testibumin 12 ; , plasminogen [36S]methionineincorporation and immunoprecipita- activator 13 ; , macroglobulin 141, inhibin E ; , growth tion demonstrated that Sertoli cells synthesize and se- factor s ; 16 ; , and many others 17, 18 ; . Many of these crete these proteins in vitro. Because they seem not to proteins are also synthesized and secreted by hepatocytes or have been isolated previously, are concentrated in and other cells into the systemic circulation. These macromolesynthesized by the testes, and are structurally related, cules serve as carriers, protease inhibitors, hormones as well we propose that CMB-22 and CMB-23 be designated as a variety of unknown functions. We have previously identestin I and testin 1 , respectively. The distribution of tified two Sertoli cell secretory proteins designated CMB-22 1 these proteins in biological fluids were compared with and CMB-23 18 ; . In thepresent studywe report their unique those of testibumin and rat androgen binding protein NH2-terminal amino acid sequences and propose that these rABP ; , two other Sertoli cell proteins. The results proteins be named testin I and testin 11. The secretion of suggest that testins, unlike testibumin and rABP, are these novel proteins appearsto be suppressed by the presence not transported to the epididymis. Although the of germ cells. amount of testins secreted by Sertoli cells in vitro is similar to that of testibumin and rABP, the concentraEXPERIMENTALPROCEDURES tions in testis and rete testisfluid are several ordersof Biochemicals-'251-Labeled Bolton-Hunter reagent N-succinimimagnitude less than that of testibumin and rABP. specific activity These observations suggest that thesecretion of these dyi 3- 4-hydr0xy-5- '~~I]iodophenyl ; propionate; proteins in vivo might be suppressed by germ cells. The 3056-3324 Ci mmol ; was obtained from ICN Radiochemicals. L-[3sS] fact that 10 times more testins are secreted by tubules Methionine specific activity 1117-1151 Ci mmol ; , methyl-14C-methylated myosin M , M, from immature rats thanby those from adult rats and 69, 000 ; , ovalbumin 200, 000 ; , phosphorylase b M , 97, 000 ; , BSA and Mr45, 000 ; , carbonic anhydrase M , 30, 000 ; , that there is an increase in the testicular content of EnlighteningTMautoradiography enhancer were obtained from DU testins following a single dose of busulfan, which de- Pont-New England Nuclear. Tris, EDTA, Ches, bis-Tris, andphenylpleted the germ cells from the seminiferous epithelium, methylsulfonyl fluoride were obtained from Aldrich. Sodium deoxysupports this hypothesis. Thus, the secretion of testins cholate, BSA RIA grade ; , and bacitracin were from Sigma. Triton by Sertoli cells appears to be tightly coupled to the X-100, sodium dodecyt sulfate SDS ; , 2-mercaptoethanol, glycine, presence of germ cells; there is an inverserelationship and N, N'-diallyltartardiamide werefrom Bio-Rad. DL-Methionine between the amount of testins in the testis and the was from United States Biochemical Corp. Acrylamide, N, N'-methStaphylococnumber of germ cells. These results suggest that testins ylene-hisacrylamide, immunoprecipitin formalin-fixedmarkers were cus a u r cells ; and prestained high molecular weight are unique testicular proteins that can be usedto study from Bethesda Research Laboratories. Sertoli cell-germ cell interactions in the seminiferous Animals-Male Sprague-Dawley rats at 18 days of age were obepithelium.

Causing bacteria. The stickier the snack candy, raisins, dried apricots ; and the more frequently it is ingested, the likelier there will be decay. Use of toothbrush, floss and mouthrinse Advise the patient to floss and to brush with an overthe-counter fluoride toothpaste and to rinse daily with an over-the-counter nonalcohol mouthrinse containing 0.05% NaF. Dentists treating high-risk patients should be aware of the newest protocols for caries management: Fluoride treatments Prescribe a 5, 000 parts per million fluoride toothpaste e.g., Prevident 5000 Plus, ControlRx ; . Rx Prevident Disp 4.2 oz 1 tube ; Sig Use to brush teeth 2 times per day, especially before going to sleep at night In the dental office, apply fluoride varnish e.g., Cavity Shield, Duraflor ; to the teeth twice a year. Antimicrobial products Have the patient rinse and brush his or her teeth with iodine for 1 minute once every 2 months ensure the patient has no allergies to iodine or shellfish ; . Rx Betadine Solution Disp 8 oz bottle Sig Once every 2 months, rinse and brush with 1 teaspoon of Betadine Solution in mouth and around teeth, then spit out. or Have the patient rinse and brush his or her teeth with chlorhexidine once a day for 2 weeks every 3 months. Rx Chlorhexidine 0.12% Disp 16 oz. Sig Every 3 months, rinse and brush with 1 tablespoon of the chlorhexidine solution in mouth for 1 minute once a day for 2 weeks and fluoride. Studying these patients for 30 days. They report an improvement in phrenic nerve function in a few of their patients, but most patients had not improved by 30 days. It will be interesting to hear the results as the authors follow these patients for a longer period of time to document the electrophysiologic recovery ofthe phrenic nerve. It may be that the phrenic nerve will not recover in some patients or, if it does, the underlying lung will be and not "trapped"movefunction will this be restored. As we forward in era of managed care, outcomes will come under more se and formoterol.

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