Renal targeting of arginine-vasopressin by modification with carbohydrates at the tyrosine side chain.

To extend the utility of a renal targeting system using carbohydrate derivatives, we investigated the in vivo tissue distribution in rats of arginine-vasopressin (AVP) derivatives modified at the phenolic hydroxy group of tyrosine by linking it to some sugars, namely D-glucose, D-galactose, D-mannose and L-fucose, via an octamethylene group. The glycosyl and mannosyl derivatives of AVP exhibit renal-selective distribution in vivo. In addition, the glucosyl and mannosyl derivatives exhibited specific binding to the kidney microsomal fraction in vitro. Modification with D-glucose or D-mannose at the tyrosine side chain is a suitable methodology for renal targeting, as well as at N-terminal amine.

Specific renal delivery of sugar-modified low-molecular-weight peptides.

To develop a novel delivery system for peptides involving sugar modification, Arg-vasopressin (AVP) was modified by linking it to a variety of sugars via an octamethylene group and the subsequent tissue uptake by rats was then monitored after administration by i.v. injection. The glucosyl, mannosyl, and 2-deoxyglucosyl derivatives of AVP exhibited selective renal uptake. These derivatives were found to be distributed in the proximal tubules of the renal cortex. In addition, they exhibited specific binding to the kidney microsomal fraction in vitro (Kd = approximately 60 nM), suggesting that they are taken up by a specific recognition mechanism located in the kidneys. From the results of the uptake study of glucosyl derivatives, the following points are clear: 1) renal uptake in vivo becomes saturated with increasing dose, and the Km from the uptake study is almost the same as the Kd obtained in the binding assay in vitro and 2) because the renal first-pass uptake extraction is about 70% at a low dose (10 nmol/kg), there is an effective mechanism for uptake from blood. Furthermore, glucosyl and mannosyl derivatives of oxytocin, a neutral peptide, unlike AVP that is basic, also have high renal uptake clearances. Thus, the renal uptake may not be dependent on derivatives having a cationic nature. We conclude that there is a novel transport mechanism in the kidneys that can be used for the specific renal delivery of glycosylated peptides.

Who's up first? Testing for order effects in structured interviews using a counterbalanced experimental design.

A growing body of research suggests that, apart from the wording of specific questions, various aspects of the interview process itself may affect the reliability of information provided by research participants. To examine whether the order of presentation of specific diagnostic modules affects the likelihood of subjects' yes/no responses within the Diagnostic Interview Schedule for Children (DISC), the authors used a counterbalanced design, presenting two DISC diagnostic modules to children and their parents in standard or reversed order. Results indicate that the order of module administration exerts effects on the total numbers of symptoms endorsed, level of impairment, and the likelihood of meeting diagnostic criteria, regardless of whether the information is provided by parent or child respondents. Future child and adult assessment measures should take these difficulties fully into account through novel approaches to instrument design and interview procedures.

Synthesis and antidiuretic activities of novel glycoconjugates of arginine-vasopressin.

Arginine-vasopressin (AVP) was acylated with various acyl azides (2a-j) in pH 9.1 buffer to give AVP derivatives (11a-j) modified at the tyrosine side chain with a carbohydrate via a spacer arm. Glycoconjugates of AVP modified at the N-terminal amide (12a-e) were also synthesized from AVP and carboxylic acids (3a-e) using dicyclohexylcarbodiimide and 1-hydroxybenzotriazole as coupling agent. Analogues (11a-j) exhibited greater in vivo antidiuretic activity than AVP. AVP and glycoconjugates (12a-e) were stable in rat plasma. On the other hand, glycoconjugates (11a-i) were found to readily convert to AVP according to first order kinetics. Hence, 11a-j are considered to be prodrugs of AVP.

Scales, diagnoses, and child psychopathology: II. Comparing the CBCL and the DISC against external validators.

Using a multimethod multistage screening procedure, the authors interviewed 201 parents and their children with the Diagnostic Interview Schedule for Children (DISC 2.1). In addition, parents completed the Child Behavior Checklist (CBCL) and other survey measures, while their children completed self-report scales. Receiver operating characteristic (ROC) analyses were done to determine optimal cutpoints on the CBCL, referenced to DISC diagnostic "caseness." DISC diagnoses, DISC "stem" symptoms, CBCL scores, and CBCL ROC-outpoints were compared against "external validators," in order to determine the comparative advantages of each approach for assessing child psychopathology. Overall findings suggest that the controversies about "best" assessment strategies may be artificial: When both assessment approaches are compared using similar methods, they are reasonably comparable. However, highly specific diagnostic categories may show fewer relationships with external validators and may therefore need more systematic validational studies.

The exceptional family member program: perceptions of active duty enrollees.

Fifty-four soldiers who had a family member enrolled in the Army Exceptional Family Member Program (EFMP) were surveyed as to their experiences and perceptions of the program. A high percentage of these soldiers reported satisfaction with treatment of the EFMP family member. They generally perceived the EFMP positively and were very supportive of the program. The majority of these soldiers also reported that their units were supportive of them and that they encountered no significant obstacles to career advancement.

Prevalence of mental disorder in military children and adolescents: findings from a two-stage community survey.

OBJECTIVE: Because previous reports have suggested that children of military families are at greater risk for psychopathology, this study examines the levels of psychopathology in an epidemiological community sample of military children all living on a military post. METHOD: Standardized psychopathology rating scales and a structured diagnostic interview (the Diagnostic Interview Schedule for Children [DISC], version 2.1) were used in a multimethod, multistage survey; 294 six- to seventeen-year-old military children and their parents participated in the study. RESULTS: Parent- and child-administered structured DSM-III-R DISC interviews indicated that children's levels of psychopathology were at levels consistent with studies of other normal samples. In addition, parents' and children's symptom checklist ratings of children were at national norms, as were parents' ratings of their own symptoms. CONCLUSIONS: Overall results do not support the notion that levels of psychopathology are greatly increased in military children. Further studies of military families should address the effects of rank and socioeconomic status, housing, and the current impact of life stressors on the parents as well as the children, in order to avoid drawing erroneous conclusions about parts or all of the military community.

Soldier functioning under chronic stress: effects of family member illness.

The psychological functioning of soldiers with a handicapped child in the family were compared with that of soldiers without such a child through a survey of 443 soldiers. Self-report questionnaires were utilized to measure depressive symptoms, martial adjustment, social supports, stressful life events, military satisfaction, military performance, and coping. Differences between the 147 soldier-parents with a handicapped child and those without were examined using one-way analyses of variance. The results indicated that soldier-parents with a handicapped child showed significantly higher depressive symptoms, including lower scores on coping, less favorable perception of their military skills and abilities, and more pessimistic attitudes about their long-term military career options, than did the comparison group. Differences in marital satisfaction were not found. Also, perceived social supports played a significantly greater role in buffering the effects of stress on marital adjustment among families with a handicapped child than among those without.

Synthesis of artificial glycoconjugates of arginine-vasopressin and their antidiuretic activities.

Arginine-vasopressin (AVP) derivatives modified at the glutamine side chain amide with carbohydrate via an alkylene spacer (1a--d) were synthesized from new glycosylated glutamine derivatives (3a--d) by solid-phase synthesis. Glycoconjugates of AVP modified at the C-terminal amide (2a--d) were also synthesized from vasopressionic acid. All of them exhibited antidiuretic activity.

The Exceptional Family Member Program: views of unit leaders.

This survey indicated that unit leaders perceived the Army Exceptional Family Member Program (EFMP) positively and considered the program beneficial to the Army, the soldiers, and their families. Furthermore, the unit leaders considered the EFMP soldiers to be well accepted in their units and their duty performance to be comparable to that of non-EFMP soldiers.

Scales, diagnoses, and child psychopathology: I. CBCL and DISC relationships.

OBJECTIVE: To clarify the relationship between scales and structured diagnostic interview diagnoses, the authors used a two-stage screening method to study 201 military families with one or more children ages 5 to 17. METHOD: Parents and children were interviewed with the Diagnostic Interview Schedule for Children (DISC 2.1); parents also completed the Child Behavior Checklist (CBCL) while the children completed other self-report symptom scales. RESULTS: Results indicate only a modest ability of scales to discriminate among discrete DISC-derived DSM-III-R diagnoses. Inclusion of diagnostic information from both parents and children resulted in more diagnoses than from either informant alone, and the additional diagnoses consisted mostly of internalizing disorders contributed by child-derived DISC information. In general, correlations were larger between scales and diagnoses within the same informant (regardless of diagnostic construct) than across informants (but within the same diagnostic construct). Child self-report measures tended to outperform the CBCL as screeners against the overall "caseness" criterion on the DISC. However, child self-report scales were relatively nonspecific and showed little ability to selectively identify internalizing disorders such as anxiety and/or depression. Compared with single informant diagnoses, combined-informant diagnoses were generally superior in demonstrating broader relationships to both parent and child symptom scales. CONCLUSIONS: Additional research is needed in order to build careful crosswalks between the various approaches to assessing childhood psychopathology, to decide on optimal rules for combining information to establish diagnoses, and to validate the currently available assessment alternatives.

Synthesis of omega-(methoxycarbonyl)alkyl and 9-(methoxycarbonyl)-3,6-dioxanonyl glycopyranosides for the preparation of carbohydrate-protein conjugates.

omega-(Methoxycarbonyl)alkyl glycopyranosides of D-mannose having C4, C7, C9, C12, and C15 carbon chains, L-fucose and 2-acetamido-2-deoxy-D-mannose having C7 and C9 carbon chains, D-xylose and 2-acetamido-2-deoxy-L-fucose having a C9 carbon chain, and 9-(methoxycarbonyl)-3,6-dioxanonyl glycopyranosides of D-mannose, 2-acetamido-2-deoxy-D-mannose, and L-fucose were synthesized as intermediates for coupling to human serum albumin in order to examine the effect of chain length and hydrophobicity of the spacer arm on the binding specificity of lectins. 8-(Methoxycarbonyl)octyl glycosides of beta-D-Man-(1----2)-alpha-D-Man, alpha-D-Man-(1----2)-alpha-D-Man, alpha-D-ManNAc-(1----2)-alpha-D-Man, beta-D-GlcNAc-(1----2)-alpha-D-Man, and their 6-O-positional isomers, beta-D-Man-(1----6)-alpha-D-Man, alpha-D-Man-(1----6)-alpha-D-Man, alpha-D-ManNAc-(1----6)-alpha-D-Man, and beta-D-GlcNAc-(1----6)-alpha-D-Man, were also synthesized.

Effects of subacute treatment with cocaine on activities of N-demethylase, UDP-glucuronyltransferase and sulfotransferase in WKY and SHR rat liver--sex and strain differences.

The effects of subacute treatment with cocaine on activities of cocaine N-demethylase, UDP-glucuronyltransferase (GT) toward 4-nitrophenol and phenolphthalein and sulfotransferase (ST) toward androsterone and 4-nitrophenol in livers from Wistar Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) were investigated. Hepatic metabolism of cocaine was different between the sexes (with males having higher N-demethylase activity) and the strains (with WKY rats having higher activity). The effects of subacute cocaine administration on the activity of cocaine N-demethylase were also sex- and strain-related. Whereas cocaine administration increased activity of hepatic N-demethylase in both female strains, it decreased activity in male WKY and had no effect on activity in male SHR. Sex and strain-related as well as cocaine-induced differences were also found in activities of hepatic GT toward 4-nitrophenol and phenolphthalein as well as in activity of hepatic ST towards andersterone and 4-nitrophenol. These results suggest that some of the individual variation in the effects of cocaine may be due to sex and genetic differences in the hepatic metabolism of cocaine and/or in sexually and/or/genetically-determined differences in how cocaine affects hepatic metabolism of other xenobiotics.

Effects of cold stress on brain regional calcium content in rats and mice.

The calcium levels of striatum, cerebellum, cortex, medulla and diencephalon from rat brain were significantly reduced after 30 minutes of cold exposure at 8 degrees C even though body temperatures were no different from pre-cold exposure values. Calcium content remained significantly depressed in striatum, cerebellum and diencephalon after 2 hours of cold exposure, at which time body temperatures were significantly depressed. A similar effect of short-term cold stress on calcium content in these brain regions from mice was observed. These results are consistent with a role of calcium in the maintenance of temperature and in adaptive responses to cold environments.

Sensitivity difference to hepatotoxicity of cocaine in spontaneously hypertensive and Wistar Kyoto rats.

Experiments were conducted to determine the hepatic damage of cocaine in spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto (WKY) rats in terms of serum glutamic-oxaloacetic transaminase (SGOT) activity, liver weight/body weight ratio and hepatic microsomal enzyme activity, i.e., N-demethylase activity or UDP-glucuronyltransferase (GT) activity. In subacute experiments, 2, 4 and 10 daily cocaine treatments elevated the level of SGOT activity and reduced the liver weight/body weight ratio in SHR rats. The ethylmorphine N-demethylase activity and the cocaine N-demethylase activity in SHR rats were significantly greater (31% and 26%, respectively) than those in WKY rats. Ten daily treatments with cocaine diminished the ethyl morphine N-demethylase activity and the cocaine N-demethylase activity in SHR and WKY rats. However, attenuation of 4-nitrophenol GT activity was only observed in SHR rats. In acute experiments, a single dose of cocaine, 40 mg/kg, elevated the SGOT activity in SHR rats and reduced the 4-nitrophenol GT activity in SHR rats, but it did not affect the activities of SGOT and 4-nitrophenol GT in WKY rats. A higher dose of cocaine, 60 mg/kg, elevated the SGOT activity and reduced cocaine N-demethylase activity and 4-nitrophenol GT activity in both SHR and WKY rats. The present studies suggest that N-demethylation of cocaine plays an important role in the hepatotoxicity of cocaine in animals.

Relationship between the neurotoxicities of Soman, Sarin and Tabun, and acetylcholinesterase inhibition.

Acetylcholinesterase (AChE)-induced chewing movements, tremors, convulsions and hind limb abduction at doses of 50-85% LD50 in rats were monitored in order to determine whether the severity of these different signs would correlate with brain AChE levels and the time course of such a relationship. 30 min after subcutaneous (s.c.) injection of Soman, the intensities of toxic signs were significantly correlated with the degree of striatal AChE inhibition. In the case of Sarin, the corresponding r-values were not significant except for tremors. For Tabun-induced chewing, tremor and hind-limb abduction, the r-values were significant. The neurotoxicity was most intense between 15 min to 2 h after treatment, but at 2 or 6 h, the r-values were well below 0.5. The inhibition of brain AChE was maximal by 30 min and was still high at 24 h.

Selective inhibitory effect of organophosphates on UDP-glucuronyl transferase activities in rat liver microsomes.

The effects of acute and subacute administration of diisopropylfluorophosphate (DFP) and acute administration of Soman, Sarin and Tabun on UDP-glucuronyltransferase (GT) activity towards 4-nitrophenol, 4-methylumbelliferone, phenolphthalein and testosterone in rat liver microsomes were investigated. Twenty-four hours after a single injection of DFP, the activity of GT towards 4-nitrophenol and 4-methylumbelliferone was inhibited, and the inhibitory effect continued for 3 days. The activity had recovered by 7 days after injection. The activity of GT towards phenolphthalein and testosterone was not affected at any time after injection. Soman, Sarin and Tabun showed the same effect as DFP after a single injection. After daily DFP injections, the activity of GT towards 4-nitrophenol and 4-methylumbelliferone was decreased to the same level as found following acute treatment with DFP. The in vitro addition of DFP to liver microsomes did not affect GT activity towards 4-nitrophenol. It is suggested that these changes are not due to a direct effect of DFP. Furthermore, the effects of two enzyme inducers on GT activity in the presence and absence of DFP were investigated. In the 3-methylcholanthrene (MC) pretreatment group, DFP inhibited only the GT activity towards 4-nitrophenol and 4-methylumbelliferone. On the other hand, in the phenobarbital (PB) pretreatment group, DFP did not inhibit the GT activity towards 4-nitrophenol and 4-methylumbelliferone. It was also demonstrated that MC pretreatment increased the mortality in the DFP-treated rats but that PB pretreatment suppressed it. These results suggest that DFP and other organophosphorus agents may be useful agents for studies on the heterogeneity of GT.

Acute effects of Soman, Sarin, and Tabun on cyclic nucleotide metabolism in rat striatum.

Rats were injected sc with 120 micrograms/kg Soman, 120 micrograms/kg Sarin or 240 micrograms/kg Tabun. At 15 min, 2 h, or 6 h after administration, animals were decapitated along with saline-treated controls, and striatal activities of nucleotide cyclases and phosphodiesterases and striatal cyclic nucleotide levels were determined. All three agents had two similar effects on rat striatal cyclic nucleotide systems: they all increased cyclic GMP levels 15 min after their administration, and they all decreased guanylate cyclase activity 2 h after administration. There were also some different effects elicited by these three organophosphorus compounds. Different effects of Soman and Sarin seem to be mainly due to their different potencies, which in turn influence the time course of their actions. Tabun is quite different from Soman and Sarin in several respects: it rarely causes convulsions at sub-lethal doses, it has no effects on striatal cyclic AMP levels, and it affects enzyme activities 6 h after its administration. These differences may be due to the presence of cyanide instead of fluoride in its structure: i.e., this may be responsible for the different effects of Tabun on striatal cyclic nucleotide systems, and perhaps other biochemical effects. These results also indicate that other neurotransmitter systems, in addition to the cholinergic system, may be involved in organophosphate-induced toxicity.

A survey of adolescent military family members' self-image.

The Offer Self-Image Questionnaire was administered to 135 adolescent military family members, ages 13 through 18. The data suggest that the usual demands placed on the military family do not deter the adolescent from developing a healthy self-image.

Recognition and treatment of acute alcohol withdrawal syndromes.

The alcohol withdrawal syndromes are generally self-limited processes from which spontaneous recovery can be anticipated. To achieve this outcome, the various types of withdrawal must be managed in such a way as to prevent the occurrence of life-threatening situations. This begins with a good initial evaluation, followed by the appropriate pharmacologic and behavioral steps to control the severity of withdrawal symptoms and to manage complications. Once the withdrawal process is completed, the patient can then be entered into a long-term treatment program.