A statistical investigation of the time-sampling methods in studying primate behavior

Two commonly used time-sampling techniques in studying animal behavior, namely, fixed interval time point technique and fixed interval time span technique have been investigated, in which their statistical properties and the estimators for frequency and duration have been discussed. Three simple numerical examples have been used to illustrate the calculation of estimates. Finally, a sketch of a stochastic approach to the problem and the resultant estimators are presented, in which all the possible transitions are considered. Therefore, both total frequency and duration of a certain behavior can be estimated by summing up the estimators during each fixed intervening interval with only two end-points being observed.     

Effects of cAMP,theophylline, imidazole,and 4-(3,4-dimethoxybenzyl)-2-imidazolidone on the leaf movement rhythm of Trifolium repens—a test of the cAMP-hypothesis of circadian rhythms

The period length of the leaf movement rhythm of Trifolium repens L. is lengthened by continuously offered cAMP (0.5–1.0 mol m^-3) and theophylline (0.5–4 mol m^-3). At the higher concentrations this effect is more pronounced and the rhythm damps out faster. Imidazole (0.5 and 1 mol m^-3) has no effect on the period length; however, after 5 mol m^-3 the rhythm is abolished. Offered as 4 h pulses the resulting phase response curves for cAMP and imidazole are similar and show delays of up to 4 h during the day position of the leaves. Theophylline pulses lead to delays of up to 5 h during closure and advances of up to 3 h during opening. No phase shift is brought about by 4-(3,4-dimethoxybenzyl)-2-imidazolidone. The results do not support the cAMP-model of the circadian clock which has been proposed by Cummings (J. theor. Biol. 55, 455–470; 1975). The effect of the substances tested could, however, be based upon influences on the transport of Ca²⁺.Abbreviations ATP adenosine triphosphate - cAMP cyclic adenosine 35 monophosphate - AMP adenosine 5 monophosphate - AC adenyl cyclase - PDE phosphodiesterase - LL continuous light     

Stability of glucose oxidase and catalase adsorbed on variously activated 13X zeolite

The use of 13X zeolite (0.1-0.4-mm granules), treated with 2N and 0.01N HCI, 0.01M citric acid, 0.1M citric-phosphate buffer (pH 3.6), and in untreated form to adsorb glucose oxidase of fungal origin and microbial catalase was examined. Physicochemical analysis of the support demonstrated that its crystalline structure, greatly altered by the HCl and buffer, could be partially maintained with citric acid. The specific adsorption of the enzymes increased with decreasing pH and proved to be considerable for all the supports. The stability with storage at 25 degrees C is strictly correlated with the titrable acidity of the activated zeolite expressed as meq NaOH/g and with pH value of the activation solution. It proved to be lower than 55 h for both enzymes if adsorbed on zeolite treated with 2N HCl, and 15-fold and 30-fold higher for glucose oxidase and catalase adsorbed, respectively, on zeolite treated with the 0.1M citric-phosphate buffer and 0.01M citric acid. The specific adsorption of glucose oxidase and catalase was, respectively, 1840 U/g at pH 3.0 and 6910 U/g at pH 5.0. Their half-life at 25 degrees C with storage at pH 3.5 for the former and at pH 5.0 for the latter was 800 and 1560 h vs. 40 and 110 h for the corresponding free enzymes.     

Interaction of glutathione transferase from horse erythrocytes with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole

7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole reacts with two thiol groups of the dimeric horse erythrocyte glutathione transferase at pH 5.0, with strong inactivation reversible on dithiothreitol treatment. The inactivation kinetic follows a biphasic pattern, similar to that caused by other thiol reagents as recently reported. Both S-methylglutathione and 1-chloro-2,4-dinitrobenzene protect the enzyme from inactivation. Analysis of the reactive SH group-containing peptide gives the sequence Ala-Ser-Cys-Leu-Tyr, identical with that of the peptide that contains the reactive cysteine 47 of the human placental transferase. In the presence of glutathione, the enzyme is not inactivated by this reagent, but it catalyzes its conjugation to glutathione. At higher pH values, 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole reacts with 2 tyrosines/dimer and lysines, as well as with cysteines. Reaction with lysine seems essentially without effect on activity; whether the reactive tyrosines are important for activity could not be determined using this reagent only. However, 2 tyrosines among the 4 that are nitrated by tetranitro-methane are important for activity.     

Redox forms of human placenta glutathione transferase

Human placenta glutathione transferase (EC 2.5.1.18) pi undergoes an oxidative inactivation which leads to the formation of an inactive enzymatic form which is homogeneous in several chromatographic and electrophoretic conditions. This process is pH dependent, and it occurs at appreciable rate in alkaline conditions and in the presence of metal ions. Dithiothreitol treatment completely restores the active form. -SH titration data and electrophoretic studies performed both on the oxidized and reduced forms indicate that one intrachain disulfide is formed, probably between the two faster reacting cysteinyl groups of each subunit. By the use of a specific fluorescent thiol reagent the disulfide forming cysteines have been identified as the 47th and 101th residues. The disulfide formation causes changes in the tertiary structure of this transferase as appears by CD, UV, and fluorometric analyses; evidences are provided that one or both tryptophanyl residues of each subunit together with a number of tyrosyl residues are exposed to a more hydrophilic environment in the oxidized form. Moreover, electrophoretic data indicate that the subunit of the oxidized enzyme has an apparent molecular mass lower than that of the reduced transferase, thereby confirming structural differences between these forms.     

Sulfur-containing cyclic ketimines and imino acids. A novel family of endogenous products in the search for a role.

Aminoethylcysteine, lanthionine, cystathionine and cystine are mono-deaminated either by L-amino-acid oxidase or by a transaminase exhibiting the properties described for glutamine transaminase. The deaminated products cyclize producing the respective ketimines. Authentic samples of each ketimine were prepared by reacting the appropriate aminothiol compound with bromopyruvate, except cystine ketimine which required the interaction of thiopyruvate with cystine sulfoxide. Reduction of the first three mentioned ketimines with NaBH4 yields the respective derivatives with the saturated rings of thiomorpholine and hexahydrothiazepine. The same reduction is carried out enzymically by a reductase extracted from mammalian tissues. Properties of the members of this family of compounds are described. Gas chromatography followed by mass spectrometry permits the identification of most of these products. HPLC is very useful for the determination of the ketimines by taking advantage of specific absorbance at 380 nm obtained by prior derivatization with phenylisothiocyanate. Adaptation of these and other analytical procedures to biological samples disclosed the presence of most of these compounds in bovine brain and in human urine. By using [35S]lanthionine ketimine as a representative member of the ketimine group, the specific, high-affinity, saturable and reversible binding to bovine brain membranes has been demonstrated. The binding is removed by aminoethylcysteine ketimine and by cystathionine ketimine indicating the occurrence in bovine brain of a common binding site for ketimines. The reduced ketimines are totally ineffective in competing with [35S]lanthionine ketimine. Alltogether these findings are highly indicative for the existence in mammals of a novel class of endogenous sulfur-containing cyclic products provided with a possible neurochemical function to be investigated further.     

Migration of the Fungal Protein Cryptogein within Tobacco Plants

Cryptogein (CRY), a protein secreted by Phytophthora cryptogea, causes necrosis on tobacco (Nicotiana tabacum) plants at the site of application (the stem or the roots) and also on distant leaves. Autoradiography of plantlets after root absorption of radioiodinated CRY demonstrated a rapid migration of the label to the leaf lamina via the veins. Using an anti-CRY antiserum, a CRY-related antigen was detected in the stem and leaves of CRY-treated plants at a distance from the site of application. This antigen had the same molecular weight as CRY and was detected in the leaves as early as 1 hour after stem treatment, i.e. long before necrosis was detectable. The antigen was also detected in plants inoculated with P. cryptogea. The distant location of the necrosis induced by the fungus or by CRY can be ascribed to the migration of this protein, which is toxic to tobacco cells. It is proposed that CRY, which also elicits defense reactions in tobacco, might contribute to the hypersensitive response of tobacco to P. cryptogea.     

Parasitic helminths of the cecum and colon of equidae in Italy]

Intestinal helminths from coecum and colon were studied in 93 equidae including 40 horses, 36 donkeys and 17 mules. A total of 38 species, 36 nematodes and 2 cestodes, were identified as follows: 1) Triodontophorus serratus, 2) Triodontophorus brevicauda, 3) Strongylus equinus, 4) Strongylus edentatus, 5) Strongylus vulgaris, 6) Cyathostomum tetracanthum, 7) Cyathostomum coronatum, 8) Cyathostomum labiatum, 9) Cyathostomum labratum, 10) Cyathostomum alveatum, 11) Cyathostomum pateratum, 12) Cyathostomum catinatum, 13) Cyathostomum sagittatum, 14) Cylicodontophorus bicoronatus, 15) Cylicocyclus radiatus, 16) Cylicocyclus auriculatus, 17) Cylicocyclus elongatus, 18) Cylicocyclus nassatus, 19) Cylicocyclus insigne, 20) Cylicocyclus leptostomus, 21) Cylicostephanus calicatus, 22) Cylicostephanus poculatus, 23) Cylicostephanus minutus, 24) Cylicostephanus longibursatus, 25) Cylicostephanus hybridus, 26) Cylicostephanus goldi, 27) Cylicostephanus ornatus, 28) Cylicostephanus skrjabini, 29) Poteriostomum ratzii, 30) Gyalocephalus capitatus, 31) Parascaris equorum*, 32) Probstmayria vivipara, 33) Draschia megastoma*, 34) Habronema muscae*, 35) Habronema majus*, 36) Setaria equina*, 37) Anoplocephala perfoliata, 38) Paranoplocephala mamillana. The asterisked species are those not usually localized in the examined material. The most frequent parasites were found in all three hosts. Species 1, 4, 6, 9, 10, 12, 21, 22, 30 and 35 showed significant differences in prevalence between horses and donkeys, the mule generally having intermediate values. Multiple infections and total worm burden appear to decrease in older animals (> 15 years). Parasite associations occur mostly at random as expected from the values of the respective total prevalences. Some significant excesses on expected values were recorded but not significant deficits. The total worm burden increases with the number of parasite species and the increase appears to follow an exponential pattern.