Hydroquinone derivatives and monoterpenoids from the Neotropical liverwort Plagiochila rutilans.

The structure of a prenylbenzene derivative isolated previously from a Cuban specimen of the liverwort Plagiochila rutilans is revised to 2-methoxy-6-prenylhydroquinone. The hydroquinone was observed as a prominent component of the NMR and GC-MS fingerprints of five recent specimens of the liverwort from Bolivia, Brazil and Costa Rica. The corresponding quinone was observed as a minor component. Two new methylated derivatives of the hydroquinone were observed as prominent components in one specimen from Bolivia; these were isolated, characterized, and their structures elucidated as 2-methoxy-1-O-methyl-6-prenylhydroquinone and 2-methoxy-4-O-methyl-6-prenylhydroquinone using 1H NMR spectroscopy. The liverwort has a strong peppermint-like odour that is caused by the presence of several menthane monoterpenoids, including notably pulegone, menthone, isomenthone. terpinolene and limonene. One of the Costa Rican specimens contained considerable amounts of the new lactone 3,7-dimethyl-2,6-octadien-1,6-olide as the principal monoterpenoid in place of pulegone. Two Costa Rican specimens distinguished morphologically as Plagiochila standleyi (a taxon closely related to P. rutilans and reduced elsewhere to a variety of that species) are characterized by large amounts of 3-hydroxy-4'-methoxybibenzyl. P. standleyi was also reported to have a peppermint-like odour in the field. Menthane monoterpenoids were again responsible but in this case the major components were limonene, beta-phellandrene, alpha-terpinene and the endoperoxide ascaridole.

Effect of exogenous somatotropin on hematological variables of lactating cows and their offspring.

Eighty-two lactating Holstein cows in their first, second, or third lactation received either one, three, or five concurrent i.m. injections of a unit dose (0.6 g) of zinc methionyl bovine somatotropin (bST) or five doses of the vehicle. Injections were administered at 14-d intervals from 60 +/- 3 d postpartum until the end of lactation or until necropsy. Thirty-eight cows were continued on the treatment for a 2nd yr. Blood samples were collected at wk -2, -1, 3, and 7 relative to the start of treatment and then every 8 wk (yr 1) or 4 wk (yr 2) thereafter. Untreated cows that were included in a survey of the resident herd were bled at wk 7 or 8, wk 10 or 11, and wk 13 or 14 of lactation and every 4 or 8 wk thereafter. Calves were bled within 72 h of birth and at approximately 5 wk of age. Most parameters associated with erythrocytes were decreased mildly in cows that were treated with bST. However, data remained within generally accepted reference ranges, and changes were not of clinical importance. Decreased hematocrit was not associated with increased hemolysis, hemodilution, or clinical anemia. No morphological lesions related to treatment were noted in the bone marrow or spleen; bST did not affect the incidence of immature cell types. Energy and protein balances did not significantly affect the hematological results of the cows. Calves generally were unaffected by bST treatment of the dam, but heavier calves had higher parameters associated with erythrocyte and lymphocyte counts than did calves with lower body weight. Exogenous bST treatment caused predictable changes in hematological parameters of dairy cows.

Effect of bovine somatotropin administered to periparturient dairy cows on the incidence of metabolic disease.

Thirty-eight dry, pregnant Jersey cows were assigned to diet and bST treatment in a 2 x 2 factorial design. During the dry period, half of the cows were fed a normal TMR (0.4% Ca; 0.3 to 0.4% P), and half of the cows were fed a high Ca TMR (1.5 to 1.6% Ca; 0.4 to 0.7% P). The high Ca diets were designed to induce milk fever and were relatively cationic (194 to 293 meq/kg) compared with the normal diets (-131 to 30 meq/kg). A standard dairy diet was fed to all cows postcalving. Cows received subcutaneous injections of either an oil-based excipient or 500 mg of bST in an oil-based excipient every 14 d from 28 d before expected calving until approximately 14 d postcalving. Peripartal bST treatment decreased the incidence of clinical mastitis, did not affect incidence of milk fever, and increased the duration, but not the incidence, of ketosis in mature Jersey cows. Blood data confirmed the clinical responses and indicated that treated cows mobilized more bone Ca than did controls, as was evidenced by increased hydroxyproline concentrations. Treatment with bST did not affect blood concentrations of 1,25-dihydroxyvitamin D, Ca, or Mg. High Ca diets increased the incidence of milk fever and downer cow syndrome compared with normal diets. The effect of bST on mastitis and milk production must be considered as preliminary given the small size of the study. Although bST treatment increased Ca mobilization, the effect was insufficient to prevent milk fever in this model.

Effect of long-term administration of a prolonged release formulation of bovine somatotropin (sometribove) on clinical lameness in dairy cows.

A matched case-control study design was used to assess the effects of long-term administration of a prolonged release formulation of bovine somatotropin (sometribove) on clinical lameness and limb lesions in dairy cows. Cows treated with sometribove for at least 2 lactations (cases) and nontreated dairy cows matched by herd, parity, age, and stage of lactation (controls) in 8 herds were evaluated for clinical lameness (as assessed by gait abnormality) and limb lesions by 2 observers, using a standardized scoring procedure at a single herd visit. Although a high proportion of the study cows were clinically lame (43%), an association was not detected between chronic administration of sometribove and prevalent lameness. Of 21 types of limb lesions identified, 2 were positively associated and 2 were negatively associated with long-term sometribove use. Superficial laceration of the tarsus (odds ratio [OR] = 2.1) and superficial swelling of the metatarsophalangeal joint (OR = 4.5) were positively associated with sometribove treatment, whereas femoral lesions (OR = 0.2) and superficial lacerations of the femur (OR = 0.14) were negatively associated with sometribove treatment.

Extended function of the corpus luteum and earlier development of the second follicular wave in heifers treated with bovine somatotropin.

Effects of recombinant bovine somatotropin (bST) on growth of the corpus luteum (CL) and development of ovarian follicles were tested. Starting at estrus (Day=0), the following treatments were administered: control (saline injected Days 0 to 19, n=5); bST[0-9] (25 mg bST injected Days 0 to 9, saline injected Days 10 to 19, n=5); bST[10-19] (saline injected Days 0 to 9, 25 mg bST injected Days 10 to 19, n=5); and bST[0-19] (25 mg bST injected Days 0 to 19, n=6). Blood was collected daily for progesterone analysis, and ultrasound examinations were performed daily for measurement of follicles and CL. Compared with the heifers treated with saline, those treated with bST had larger CL and more progesterone during the early (/=10 mm) follicles was greater (P<0.01) and largest follicles were smaller (P<0.001) in bST than in saline-treated heifers. Estrous cycle length and ovulation rate were similar for each group. In conclusion, bST increased initial development of the CL and extended its function. Furthermore, the second follicular wave was earlier with bST.

[FeS/FeS2], a redox system for the origin of life (some experiments on the pyrite-hypothesis).

The FeS/FeS, redox system, whose importance is stressed in recent theories on the origin of life, has been tested experimentally. In this paper it is demonstrated by thermodynamical calculations as well as by experiments, that cyclohexanone, which served as model compound, can be reduced by the aforementioned redox system. Reactions were carried out in methanol and DMF at 25 degrees C and at 100 degrees C. Besides products that were synthesised in both solvents, like cyclohexanethiol and dicyclohexydisulphide, special compounds were obtained in methanol and in DMF, because of the involvement of the respective solvent in the reaction. Yields of reduced compounds were lower in methanol owing to compound that hindered the reduction (cyclohexylketal). With increasing temperature and duration the amount of reduced compounds increased. Further experiments have shown that 1,1-cyclohexanedithiol is likely to be a necessary intermediate for the reduced products. The experiments give evidence to the 'pyrite hypothesis', which postulates that the FeS/FeS2, redox system was of importance for the origin of life.

Response of dairy cows to high doses of a sustained-release bovine somatotropin administered during two lactations. 2. Health and reproduction.

Eighty-two lactating Holstein cows in their first, second, or third lactation received either one, three, or five concurrent i.m. injections of a unit dose (.6 g) of zinc methionyl bST (sometribove) or five doses of the vehicle. Injections were given at 14-d intervals from 60 +/- 3 d postpartum until the end of lactation or necropsy. Thirty-eight animals were continued on treatment for a 2nd yr. Sometribove did not affect the incidence of ketosis, milk fever, tetany, or pneumonia. Digestive disorders, primarily cows going off feed, were increased by bST during yr 1 only. The incidence of lameness was increased by bST in some time frames because of an increase in the 3.0-g bST group. Lameness was not associated with treatment-specific histopathologic changes or with abnormalities in cartilage or bone. Reproductive health generally was unaffected by treatment, but delayed conception and increased incidence of abortion were noted. Incidence of cystic ovaries was unaffected by bST. Pregnancy rates were decreased during the 100-d breeding interval of yr 1 but not during the 215-d interval of yr 2. The incidence of clinical mastitis was increased by bST, primarily at the 3.0-g dose. During the 2-yr study, 0, 3, 3, and 2 cows died or became moribund on 0, .6, 1.8, and 3.0 g of bST, respectively. Health issues identified for further evaluation included lameness and clinical mastitis for the 3.0-g group and early removal from the herd and decreased reproductive performance for all bST groups. Bovine somatotropin caused no treatment-specific toxic effects in dairy cows even at 3.0 g every 14 d.

Response of dairy cows to high doses of a sustained-release bovine somatotropin administered during two lactations. 1. Production response.

This study evaluated the effect of sometribove (zinc methionyl bST) in a sustained-release formulation administered to lactating cows at concentrations up to 3.0 g every 14 d over two lactations. Eighty-two lactating Holstein cows in their first, second, or third lactation were assigned to the study. Cows received .6, 1.8, or 3.0 g of bST in one, three, or five intramuscular injections of a unit dose (.6 g) every 2 wk. Controls received five injections of the vehicle (equivalent volume to the 3.0-g treatment) every 2 wk. Injections were administered from 60 +/- 3 d postpartum until dry-off or necropsy. Thirty-eight animals were continued on treatment for a second consecutive lactation. During the 1st yr of treatment, bST increased mean 3.5% FCM by 7.2, 9.4, and 8.4 kg/d over control production (21.1 kg/d). During the 2nd yr, milk response to .6, 1.8, and 3.0 g of bST averaged 10.6, 3.6, and 4.9 kg/d over controls (24.8 kg/d). The incidence of clinical mastitis increased in the 3.0-g group relative to controls during the 2nd yr. Thus, salable FCM averaged 8.1, 9.1, and 6.2 kg/d above controls (yr 1) and 12.1, 4.7, and -2.8 kg/d (yr 2) for the .6-, 1.8-, and 3.0-g groups. Salable FCM was unaffected by mastitis at a proposed commercial dose (.6 g). Milk fat, protein, lactose, calcium, phosphorus, zinc, magnesium, and ash concentrations were unaffected by bST treatment. Calculated energy, calcium, phosphorus, and protein balances also were unaffected except for early decreases of up to 5 Mcal/d, and 40, 20, and 600 g/d, respectively, until feed intake increased. Milk serum bST concentrations greater than the assay limit of sensitivity (1 ng/ml) were routinely measurable only at doses of 1.8 and 3.0 g. Results confirmed that bST concentrations in milk serum are exceedingly small. Overall, supraphysiological doses of sometribove increased milk production with little effect on composition. No toxic effects of bST were observed.

Finishing lamb performance responses to bovine and porcine somatotropins administered by Alzet pumps.

Daily injection of ovine and bovine somatotropin (oST and bST, respectively) has been shown to improve performance and carcass quality of finishing lambs. To evaluate responses to continuously released bST and porcine ST (pST), which have 99 and 91% sequence homology with oST, respectively, finishing lambs were implanted with 2-wk Alzet pumps containing bST or pST, which was released at rates of 2 or 4 mg/d. Six-week growth rate and feed efficiency responses to bST were greater than those to pST (P less than .05). Overall feed efficiency was improved 15% and growth rate was increased 16% in lambs treated with 4 mg/d of bST compared with control lambs and neither trait was affected in pST-treated lambs. Performance responses were reflected by changes in circulating glucose, blood urea nitrogen, and insulin-like growth factor I (IGF-I) concentrations. Scatchard analysis of sera with relative binding of greater than 30% revealed that average binding capacities and affinities of pST-treated lambs were 7.0 mg/liter and 6.0 x 10(9) liters/mol, respectively, and of bST-treated lambs were .8 mg/liter and 1.3 x 10(9) liters/mol, respectively. In addition, lambs with high-capacity pST antibodies had lower 6-wk IGF-I concentrations than those of controls, suggesting that these antibodies may have been attenuating responsiveness to pST. It is concluded that continuously released bST, but not pST, improves performance of finishing lambs.

Effect of recombinantly-derived bovine somatotropin on reproductive performance of dairy cattle.

Data from 814 cows involved in five separate full lactation studies were used to assess the effect of recombinantly-derived methionyl bovine somatotropin (sometribove) on reproductive performance. Data were separated by parity (first and second or greater), route of administration (intramuscular: i.m. or subcutaneous: s.c.), length of breeding period (number of days in milk), and level of production (high vs low). Through 305 days in milk, pregnancy rates were reduced 18% in primiparous, sometribove-treated (i.m.) cows but were not significantly affected in multiparous cows. No differences were noted in either parity group when sometribove was administered subcutaneously. The level of production was more often found to be a significant factor affecting reproductive parameters than was sometribove administration. In general, length of the breeding period and level of milk production had a greater influence on reproductive performance than treatment with sometribove.

Evaluation of sometribove in a prolonged-release system in lactating dairy cows--production responses.

Primiparous (n = 105) and multiparous (n = 136) Holstein cows were used to evaluate efficacy of sometribove (n-methionyl bovine somatotropin, bST) in a dose titration study. Cows were fed TMR for ad libitum intake, were milked twice daily, and were allocated randomly within parity (1 vs. 2+) to treatments of 0, 250, 500, or 750 mg bST/14 d in a prolonged-release formulation. Subcutaneous injections commenced 60 +/- 3 d postpartum and continued throughout lactation. During a standardized treatment period of 252 d, treatment with increasing dosages of bST increased 3.5% FCM yield in a dose-dependent manner for both primiparous (2.5 kg/d, 10.2%; 3.5 kg/d, 14.3%; and 5.9 kg/d, 24.1%) and multiparous cows (3.1 kg/d, 12.1%; 3.9 kg/d, 15.2%; and 6.8 kg/d, 26.5%). Milk content of fat, protein, lactose, ash, Ca and P, and SCC were not affected by treatment. Over the 252-d treatment period, voluntary intake of energy was increased in bST-treated cows such that BW gain, body condition score, and net energy balance did not differ among treatment groups. Productive efficiency (milk per unit NEL intake corrected for BW change) over the treatment period was significantly increased in a dose-dependent manner for multiparous cows (4.1, 6.8, and 11.0%). Results demonstrated that bST administered in a prolonged-release formulation was efficacious in enhancing milk production and feed efficiency.

Gas chromatographic assay of methyltestosterone in tablets.

A simple gas chromatographic procedure has been developed for the determination of methyltestosterone in bulk powders and in tablets. Two new silyl ether derivatives of methyltestosterone have been prepared using dimethylethylsilylimidazole (DMESI) and dimethylisopropylsilylimidazole (DMiPSI). The method is accurate and selective for methyltestosterone within the concentration range 0.1-1.5 micrograms microliters-1.

Effect of acute challenge with an extreme dose of somatotropin in a prolonged-release formulation on milk production and health of dairy cattle.

Eight pregnant Holstein cows were given weekly injections of 15 g of recombinant bST over a 2-wk period for a total dose of 30 g to determine signs of acute toxicity. Cows were monitored intensively throughout the study, and samples were taken for analyses of hormones, metabolites, chemistries, hematology, and urine analytes. Animal health throughout the study was generally excellent. Mean rectal temperatures were significantly higher in treated cows (38.7 vs. 39.2 degrees C). Least squares means for 3.5% FCM production were 15.9 and 23.0 kg/d, and net energy intakes were 29.4 and 26.9 Mcal/d for control and treated cows, respectively. Somatotropin concentrations reached more than 250 ng/ml on d 10 and remained above 200 ng/ml. Insulin and glucose concentrations were also increased but returned toward baseline values. Free fatty acid concentrations were higher in treated cows, but beta-hydroxybutyrate was not affected. Most hematological measurements were unaffected except for a reduction in erythrocyte number in treated cows and decreases in hematocrit and hemoglobin, but values were within clinically normal ranges. Although cows received in 2 wk a dose that was equivalent to the amount administered during more than 2 yr of continuous use, no signs of acute toxicity to bST were observed.

Unilateral scrotal hernia repair in a ram lamb.

A hemilateral scrotal enlargement in a 5-month-old Hampshire ram lamb was diagnosed as a scrotal hernia. Surgical intervention revealed the herniation to involve only the omentum, with some testicular atrophy. The cause of this condition was thought to be trauma.

Selective reactions in the analytical characterisation of steroids by gas chromatography-mass spectrometry.

The analytical characterization, by GC-MS, of individual compounds in mixtures of steroids, such as occur frequently in biological extracts, is difficult because of the close similarities in structure and properties of many components. The improved separating power of capillary (open-tubular) columns alleviates the problem, but does not solve it fully: for example, the coincidence of retention times of two different compounds may still be virtually complete. Comparative analyses on two distinctively different phases afford one valuable application of selectivity, but may not always be feasible when costly columns are required. Comparative analyses of the sample, before and after effecting its modification by well-defined reactions, are inexpensive and are particularly when selective transformations are used. The use of the microbial enzyme cholesterol oxidase as a selective oxidant for 3 beta-hydroxysteroids (chiefly limited to 4-ene, 5-ene and 5 alpha-types) is illustrated for a model mixture of androstanols related to the boar pheromone (5 alpha, 16-androsten-3 alpha-ol). Retention regularities and changes in mass spectra enhance the reliability of identifications. An exploratory application of cholesterol oxidase in the analysis of minor "polar" sterols in human serum is reported. Most of the known minor sterols are good substrates for the enzyme, and their transformation products yield distinctive GC-MS data, as exemplified for the 7 alpha- and 7 beta-hydroxycholesterols. Another convenient and versatile selective reagent is methaneboronic acid, which yields cyclic esters of suitably constituted diols. These derivatives have shorter retention times (on "non-polar" phases) than the di-TMS ethers, chiefly by virtue of their much lower molecular weights. The mass spectra of cyclic boronates generally show clear molecular ions, also fragmentations that complement the information obtainable from the di-TMS ethers. These features are illustrated for a group of diols and triols of the 5 alpha-pregnane series.

Selective reactions in the analysis and characterization of steroids by gas chromatography-mass spectrometry.

Gas chromatography-mass spectrometry (GC-MS) is a technique especially suitable for the analysis and characterization of steroids, and its power has been extensively demonstrated. The efficacy of GC-MS is limited, nevertheless, by the fact that steroid mixtures - whether of natural origin only, or augmented by synthetic analogs - often contain similar components that are poorly distinquished. The fortuitous overlap of gas chromatographic peaks from disparate compounds also impairs the definition of retention data. Controlled modification of the sample by means of selective reactions is therefore a valuable adjunct to the application of GC-MS. Two examples are discussed: (a) the enzyme cholesterol oxidase, isolated from various microorganisms, catalyzes the oxidation of many 3 beta-hydroxy-5-enes (with concomitant isomerization) to 4-en-3-ones; 3 beta-hydroxy-5 alpha-steroids are also oxidized to the corresponding 3-ones, but other steroids (3 alpha-hydroxy- or 5 beta-isomers, etc.) are unaffected. The mild conditions required (pH 7, 30 C) are advantageous for the analysis of sensitive steroids, and the retention index increments, as well as the mass spectra of the ketones, are characteristic. The enzyme accepts as substrates a wide range of 3 beta-hydroxysteroids, tolerating oxygenation in ring B and even catalyzing the oxidation of 2-oxacholesterol to the expected lactone; and (b) Steroids possessing 1,2-diol or 1,3-diol groupings include estriols, 2-hydroxyestrone, 20,22-dihydroxycholesterols, ecdysones, brassinolide and many corticosteroids. The selective formation of cyclic derivatives can provide several analytically useful features, such as convenient retention times, moderate mass increments (24 amu for a methaneboronate), distinctive mass spectra and usually abundant molecular ions. These are exemplified for 5-pregnene-3 beta, 20,21-triols and for 20,22-dihydroxycholesterol as well as its enzymic oxidation product.

Simultaneous trace analysis of nitrous oxide and halothane in air.

A gas chromatographic method for the simultaneous analysis of halothane and nitrous oxide in operating theatre atmospheres has been developed and evaluated. The flame ionization detector is suitable for the quantitative analysis of halothane in concentrations approaching one part per million. The frequency-modulated electron capture detector is highly sensitive to nitrous oxide but we have found it to be non-linear over the range 25--1000 p.p.m. The overall reproducibility of the gas chromatographic method based on the dynamic technique of standard preparation is approximately 4%. Effective exposure of personnel to pollutant anaesthetics is assessed by the analysis of end-expired gas.

Application of the extractive alkylation technique to the pentafluorobenzylation of morphine (a heroin metabolite) and surrogates, with special reference to the quantitative determination of plasma morphine levels using mass fragmentography.

The pentafluorobenzylation of morphine and related phenolic alkaloids by extractive alkylation is described. The alkylation is performed using tetrabutylammonium as counter ion and ethyl acetate as solvent. Optimum reaction conditions are presented together with the gas chromatographic properties of the derivatives formed. The technique is applied to the quantitation of plasma morphine levels. Using morphine-d3 as internal standard mass fragmentographic analysis of morphine as its pentafluorobenzyl- and pentafluorobenzyl, mono-trifluoroacetyl derivatives is demonstrated, and a case report is presented. Quantitation to a plasma morphine level of 5 ng/ml is readily attainable.

Effect of polarity on the response of the flame ionization detector to halothane.

Prepared standards of halothane in oxygen were analysed using a Pye Series 104 gas chromatograph equipped with a heated head flame ionization detector and gas sampling valve. The adoption of a positive jet voltage expanded the linear dynamic range of the detector to cover the 0.5-9.0% by volume range, provided that a small (0.5-ml) sample loop was used.