[Retrospective analysis of dental trauma among soldiers in the Israel Defense Forces].

OBJECTIVE: Determination of the incidence, types and causes of traumatic dental incidents (TDIs) among Israel Defense Forces (IDF) soldiers. STUDY DESIGN: Dental trauma reports from all active IDF dental clinics between the years 2000-2010 were analyzed. A total of 1671 dental trauma reports were classified according to the incidence, causes and etiologies of the injuries, the number and type of traumatized teeth, and the types of dental injuries. Statistical associations between the number of trauma cases and gender, type of training of the soldier, and the month during which the injury occurred were analyzed. RESULTS: The incidence of dental injuries was 19.65 cases/10,000 soldiers/year. In basic training bases, 75.49 dental trauma cases/10,000 soldiers/year were found in comparison to 14.28 cases in all other bases. Male soldiers were 4.24 times more prone to dental injuries than female soldiers. Significantly more trauma cases occurred during the months of January and August (rate ratio 1.39 and 1.33 respectively), and significantly fewer cases occurred in July (rate ratio 0.59). The most frequent circumstances of TDIs were military training and work related injuries (29.5% and 15% respectively). The etiology of 56.3% of the injuries was trauma from blunt objects that are not a weapon. Of the injuries, 34.9% occurred as a result of trauma from the personal weapon of the soldier. Most trauma cases involved one or two injured teeth (73.2% and 20.1% respectively). Of the trauma cases, 33.8% involved the right maxillary central incisor and 32.5% involved the left maxillary central incisor. There was no significant difference between injuries on the right or left side. The most frequent type of dental injury was a crown fracture (72.8%). CONCLUSION: The risk factors for dental trauma found in this study were male soldiers during basic training in the months of January and August. Most TDIs resulted from blunt objects including personal weapons. Crown fracture was the most frequent type of injury.

[Dental care to military dogs].

Oketz is a military special unit that operates different dog species for various missions. The dogs get routine medical and dental treatments in order to maintain their health and function. The dental treatment is based on the principles of contemporary dentistry for small animals. Furthermore, these working dogs need special care due to higher risk to trauma and attrition. The dogs go through routine dental examination and prophylactic dental cleaning. Each dental procedure is performed under general anesthesia; therefore it is well planned ahead including all the pre-operative workup needed. The article presents the current concepts of dental treatment of dogs especially in respect to their activity.

Quantitative ultrasound of the tibia: a novel approach for assessment of bone status.

An ultrasound instrument has recently been developed for the diagnosis and monitoring of osteoporosis (SoundScan 2000, Myriad Ultrasound Systems Ltd., Israel). The instrument measures the speed of propagation of ultrasound waves (SOS, meters per second) along a fixed longitudinal distance of the cortical layer at the tibial shaft. Its in vivo precision is 0.25%. The performance of the SoundScan 2000 was studied in 307 Caucasian women (age range 24-87 years) who also had their bone mineral density (BMD) measured at the spine, femoral neck, and radial shaft by absorptiometric techniques. The SOS ranged from 3471-4226 m/sec (mean 3867). The standardized coefficient of variation (CV), an expression of the effective clinical precision corrected for the spread of measurements (CV/[range/mean]), was 1.6% for the tibial SOS, compared to 1.5%, 3.8%, and 4.4% for spinal, femoral, and radial BMD, respectively. Tibial SOS significantly correlated with age (r = -0.52), time since menopause (r = -0.43), height (r = 0.29), and weight (r = 0.16), as well as with BMD at the radius (r = 0.63), spine (r = 0.50), and femur (r = 0.47). After classification of bone measurements into tertiles, about 60% of the women with low tertile spinal BMD fell within the low tertile of either tibial SOS, femoral BMD, or radial BMD. The results show that measurement of tibial SOS is a precise method of assessing bone status without exposing the patient to sources of radiation.

Expression and regulation of testicular inhibin alpha-subunit gene in vivo and in vitro.

Inhibin, a gonadal peptide, selectively suppresses FSH release from the pituitary. The cDNAs coding for ovarian inhibin have been isolated and characterized. However, little is known about testicular inhibin. In this study we have isolated inhibin alpha-subunit cDNA from human testicular cDNA libraries and determined inhibin alpha-subunit mRNA levels in testes. The longest cDNA isolated from human testis was 1380 nucleotides long and contained a nucleotide sequence identical to that of human placental inhibin alpha-subunit and isolated human inhibin alpha-subunit gene, but different from human ovarian inhibin alpha-subunit in two amino acids in the signal peptide. A single 1.5-kilobase species of inhibin alpha-subunit mRNA was identified in the testes of several species. This mRNA was the same size as those in human ovary and placenta. The regulation of inhibin alpha-subunit mRNA in rat testis was next examined. The concentration of testicular inhibin alpha-subunit mRNA peaked between 20-25 days of age and gradually declined thereafter. Hypophysectomy decreased testicular inhibin alpha-subunit mRNA levels. Supplementation of hypophysectomized animals with FSH restored inhibin alpha-subunit mRNA levels to those in intact controls. By contrast, treatment with testosterone had no effect. Similarly, in Sertoli cell-enriched cultures, FSH, but not testosterone, increased inhibin alpha-subunit mRNA levels. We conclude that 1) human testicular inhibin alpha-subunit mRNA is similar to that of human ovary and placenta; and 2) inhibin alpha-subunit mRNA in Sertoli cells is regulated by FSH, but not testosterone, both in vivo and in vitro.

Solubilization and purification of rat pituitary gonadotropin-releasing hormone receptor.

Gonadotropin-releasing hormone (GnRH) receptors were solubilized from rat pituitary membrane preparations in an active form by using the zwitterionic detergent CHAPS (3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid). The solubilized receptor exhibits high affinity, saturability, and specificity. The soluble supernatant retained 100% of the original binding activity when stored at 4 or -20 degrees C in the presence of 10% glycerol. The receptors were resolved into two components on the basis of chromatography on wheat germ agglutinin-agarose. Homogeneous receptor preparation was obtained by two cycles of affinity chromatography on immobilized avidin column coupled to [biotinyl-D-Lys6]GnRH. The overall recovery of the purified receptor was 4-10% of the initial activity in the CHAPS extract, and the calculated purification -fold was approximately 10,000 to 15,000. Analysis of iodinated purified GnRH receptors by autoradiography indicated the presence of two bands, Mr = 59,000 and 57,000. This was confirmed by photoaffinity labeling of the partially purified receptors and suggests that both components can specifically bind the hormone.

Gonadotropin releasing hormone activation is mediated by dimerization of occupied receptors.

A dinitrophenyl (DNP)-derivative of a gonadotropin releasing hormone (GnRH) antagonist was prepared by chemical modification of the epsilon amino group in position 6 of [D-pGlu1,D-Phe2,D-Trp3,D-Lys6]GnRH with 1-fluoro-2, 4-dinitrobenzene. The DNP-antagonist D-pGlu-D-Phe-D-Trp-Ser-Tyr-D-Lys(N epsilon-DNP)-Leu-Arg-Pro-Gly-NH2, retained high affinity binding to the GnRH receptor of pituitary membrane preparations and exhibited antagonistic activity when assayed in cultured pituitary cells. Both antibodies against DNP and their Fab fragments were able to bind the DNP-antagonist. However, only the addition of bivalent antibodies (and not the Fab fragments) converted the DNP-antagonist to an agonist. These results suggest that divalency is a critical factor in GnRH action.

Mapping of gonadotropin-releasing hormone receptor binding site.

On the basis of the spatial conformation of gonadotropin-releasing hormone (GnRH), we have predicted that aromatic amino acids and at least one carboxyl group are involved in the recognition site of the receptor. Therefore, various specific reagents were examined for their ability to interfere with the binding of GnRH to its receptor. Pretreatment of pituitary membrane preparations with sodium periodate decreased the specific binding in a dose-dependent manner (IC50 = 0.5 mM) due to a decrease in receptor affinity. This indicated the presence of a sugar moiety in the binding site. Tryptophan is another constituent that participates in the GnRH binding site, as pretreatment of pituitary membranes with 2-methoxy-5-nitrobenzyl bromide inhibited the binding (IC50 = 0.22 mM) by decreasing receptor affinity. In addition, the native hormone conferred on the binding site a protective effect against inactivation by 2-methoxy-5-nitrobenzyl bromide. Pretreatment of membranes with p-diazobenzenesulfonic acid also inhibited the binding of 125I-Buserelin (IC50 = 0.1 mM), indicating the presence of tyrosine within or near the binding site. Pretreatment of pituitary membrane preparations with dithiothreitol also inhibited the binding due to a decrease in the binding affinity, which was accompanied by an increase in receptor number. These data suggest that there are disulfide bonds within or near the binding region. Treatment with 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide and glycine ethyl ester also prevented binding in a dose-dependent manner and implies that free carboxylic groups are involved in the binding site.(ABSTRACT TRUNCATED AT 250 WORDS)

Characterization of GnRH receptors in bovine pituitary membranes.

Bovine pituitary gonadotropin-releasing hormone (GnRH) receptors were characterized and identified utilizing a superactive GnRH analog, Buserelin [( D-Ser(t-Bu)6, des-Gly10-ethylamide]-GnRH), and a photoreactive GnRH analog, [azidobenzoyl-D- Lys6 ]-GnRH. Both analogs bind with high affinity to a single class of receptors, with apparent IC50 values of 0.5 nM and 1 nM, respectively. The binding of 125I-labeled Buserelin to pituitary membranes was inhibited, in a dose-responsive manner, by both trypsin and chymotrypsin, with the former being less effective. Neuraminidase at a concentration up to 100 micrograms/ml did not affect the binding. Lectins, such as concanavalin A and wheat-germ agglutinin, at a concentration range of 20-200 micrograms/ml had no effect on the binding, whereas soybean agglutinin at high concentrations (150 and 200 micrograms/ml) slightly inhibited the specific binding. Photoaffinity labeling of the bovine pituitary GnRH receptors resulted in the identification of two specific bands with apparent molecular weights of 60 K and 30 K daltons. The latter probably represents very low affinity binding sites. Both specific bands were sensitive to trypsin and chymotrypsin treatment but were not affected by neuraminidase treatment. These results suggest a slight difference between rat and bovine pituitary GnRH receptors.

Testicular GnRH receptors: photoaffinity labeling and fluorescence distribution studies.

Specific GnRH receptor proteins of purified rat Leydig cells and membrane preparations were identified using an 125I-labeled bioactive photoaffinity derivative of GnRH. Sodium dodecyl sulfate polyacrylamide gel electrophoresis resulted in the identification of two specific components with apparent molecular weights of 60,000 and 54,000 daltons. Fluorescent visualization of GnRH receptors in these cells, utilizing a bioactive rhodamine derivative of the hormone, indicated that the fluorescently labeled receptors were initially distributed uniformly on the cell surface and then formed clusters which subsequently internalized (at 37 degrees C) into endocytic vesicles. These processes were dependent on specific binding sites for the rhodamine-labeled peptide on Leydig cells. These findings indicate further characterization of the testicular GnRH receptors and may have important implications towards the understanding of the molecular events involved in the action of the hormone in the testis.

Receptor-mediated internalization of LHRH antagonists by pituitary cells.

A fluorescently labeled antagonist of luteinizing hormone releasing hormone (LHRH), D-pGlu-D-Phe-D-Trp-Ser-Tyr-D-Lys6-(tetramethylrhodamine)-Leu-Arg-Pro-Gly-NH2, was prepared. This peptide retained high-affinity binding to the LHRH receptor of pituitary plasma membrane preparations. The analog was able to block LHRH-stimulated LH release from pituitaries incubated in vitro, and exhibited minor agonistic activity. This rhodamine-labeled antagonist was utilized for the microscopic visualization and localization of LHRH receptors in dispersed rat pituitary cells. The fluorescently labeled receptors were initially distributed uniformly on the cell surface. The hormone-receptor complexes were redistributed after incubation at 23 degrees C and formed clusters which subsequently became internalized (at 37 degrees C) into endocytic vesicles. Addition of LHRH (10(-6) M) abolished these processes, indicating specific binding sites for the rhodamine-labeled peptide to the gonadotrope cells. A quantitative comparison of temperature-dependent internalization by iodinated LHRH agonist and antagonist revealed that both analogs were internalized to a similar extent. These findings suggest that LHRH-receptor complex internalization is related to LHRH receptor regulation.

Covalent linking of photoreactive gonadotropin-releasing hormone to gonadotropes produces a prolonged signal.

A bioactive, photoreactive derivative of gonadotropin-releasing hormone (GnRH; gonadoliberin), [azidobenzoyl-D-Lys6]GnRH, was shown to bind covalently to dispersed pituitary cells after irradiation. Approximately 7% of the total cell-associated radioactivity was covalently bound to the receptors. Photolysis of cultured pituitary cells in the presence of the photoreactive derivative resulted in persistent activation of luteinizing hormone (LH; lutropin) release. This persistent response was time dependent and concentration dependent. No increase in the basal rate of LH release was observed with cells incubated in the presence of photoreactive GnRH analog and maintained in the dark or with hormone derivatives that lack the photoreactive azido group. These results suggest that only the covalently bound cell surface receptors account for the persistent activation of LH release after photolysis.

Gonadotropin releasing-hormone receptors: photoaffinity labeling with an antagonist.

A photoaffinity antagonist of gonadotropin releasing hormone (GnRH), D pGlu-D-Phe-D-Trp-Ser-D-Lys6(N epsilon-azidobenzoyl)-Leu-Arg-Pro-Gly-NH2 (photoaffinity antagonist) was prepared by reacting [D-pGlu1, D-Phe2, D-Trp3, D-Lys6]GnRH with the N-hydroxysuccinimide ester of 4-azidobenzoic acid. The analog appeared homogeneous when analyzed by thin-layer chromatography and its photoreactivity was demonstrated by spectral changes when exposed to light. The photoaffinity antagonist retained high affinity binding to the GnRH receptor of pituitary membrane preparations and exhibited antagonistic activity when assayed in vitro in whole pituitaries. Pituitary membrane preparations were incubated with the radioactive photoaffinity GnRH antagonist and irradiated with light. Sodium dodecyl sulfate gel electrophoresis after solubilization and reduction showed the specific labeling of a single specific protein with an apparent molecular weight of 60,000 daltons. These results indicate that GnRH agonists and antagonists bind to the same receptor.

A novel method for localization of gonadotropin releasing hormone receptors.

Two 125I-labeled analogs of GnRH, [acidobenzoyl-D-Lys6]GnRH (I) and [D-Lys6]GnRH (II) were used for the localization of GnRH receptors in pituitary cells. The analogs exhibited high binding affinity to pituitary membrane preparations and after photoactivation (analog I) or cross-linking with glutaraldehyde (analog II), these analogs are bound covalently to pituitary cells. The distribution of the labeled hormones by light and electron microscopic autoradiography indicated that after exposure of pituitary cells to 125I-labeled hormones at 4 C (90 min), most of the labeled hormones were associated with the cell surface membrane, while at 37 degrees C (30 min) most of the cell-bound labeled hormones were internalized.