Measurement of Acetylcholine Release in Freely Moving Rats by Means of Automated Intracerebral Dialysis

The present study demonstrates the feasibility of measuring acetylcholine in perfusion samples collected by means of in vivo brain dialysis in the striata of freely moving rats. The output of the dialysis device was directly connected to an automated sample valve of a HPLC-assay system that comprises a cation exchanger, a post-column enzyme reactor, and an electrochemical detector. The presence of an acetylcholinesterase inhibitor (neostigmine) in the perfusion fluid was required for the detection of acetylcholine in the perfusate. Increasing concentrations of neostigmine induced increasing amounts of acetylcholine. Continuous perfusion with a fixed concentration (2 microM) of neostigmine resulted in gradually increasing amounts of collected acetylcholine over time although a considerable variation between successive samples exists. The brain dialysis technique was further validated by studying the effect of various drugs. Systemically administered atropine increased the output of acetylcholine, whereas the addition of tetrodotoxin to the perfusion fluid resulted in a complete disappearance of the neurotransmitter.     

Retrospective evaluation of gynecologic cytodiagnosis. II. Interlaboratory reproducibility as shown in rescreening large consecutive samples of reported cases

Two laboratories exchanged and rescreened a large sample of cases with cervicovaginal smears they had consecutively accessioned to examine the reproducibility of gynecologic cytodiagnosis under optimum conditions. At least a "working agreement" (diagnoses within +/- 1 category on a ten-category scale) was achieved in diagnoses of normal, benign reaction and squamous abnormality (from minimal dysplasia though invasive cancer) in 18,859 cases (96.8%), of endometrial abnormality in 21 cases (42%) and of "unsatisfactory" in 99 cases (20.7%). Larger differences occurred in greater than or equal to 30% of cases except in the categories of "normal" and "benign reaction," reaching a maximum of 82% for moderate dysplasia. Reexamining 382 cases decreased disagreement by category to the 20% to 65% range only in the five categories of dysplasia plus carcinoma in situ. Agreement was not predicated on the presence of endocervical cells or squamous metaplasia; the basis for "unsatisfactory" calls was not uniform. Comparison of the laboratories' diagnoses with referee diagnoses or, on 178 cases, with tissue diagnoses also demonstrated differences in diagnostic criteria.     

Diastereomers of thymidine 3'-O-(methanephosphonothioate): synthesis, absolute configuration and reaction with 3'-methoxyacetylthymidine under conditions of triester approach to oligonucleotide synthesis

Diastereomers of the title compound were obtained and absolute configuration was assigned by means of stereochemical correlation. Their reaction with 3'-O-methoxyacetylthymidine in the presence of triisopropylbenzenesulphonyl (4-nitro) triazole is neither chemo- nor stereo-selective and leads to diastereomeric pairs of dithymidyl (3',5')methanephosphonate and -methanephosphonothioate. Obtained results are discussed in terms of mechanism of activation of phosphodiesters under conditions known as "phosphotriester approach to oligonucleotide synthesis".     

Luteal morphology, atresia, and plasma progesterone concentrations during the reproductive cycle of two oviparous lizards, Crotaphytus collaris and Eumeces obsoletus

From ovulation to oviposition, the corpora lutea of the oviparous lizards Crotaphytus collaris and Eumeces obsoletus exhibit three stages of luteal development: 1) luteogenesis, 2) luteal maturity, and 3) luteal regression. Each stage exhibits distinct characteristics, involving changes in: 1) luteal volume, 2) nuclear diameter of cells within the luteal cell mass, and 3) thecal development. Plasma progesterone concentration is greatest during luteogenesis and is positively correlated with ovarian atresia, although atresia occurred throughout the period of gravidity. These data suggest that in these two species, the corpora lutea secrete high amounts of progesterone immediately following ovulation and exhibit morphologically distinct stages of growth and regression.     

Radioreceptor assay in checking serum concentration in long-term treatment with cis(z)-flupenthixol decanoate

24 patients have been treated with cis(z)-flupenthixol decanoate for 6-12 months. Intramuscular injections were given about every 3 weeks. Before treatment and on each day of injection the mental state was assessed by BPRS and registration of side effects was performed. Blood samples were taken 7 days after each injection and on the last day of the dosage interval. Neuroleptic activity was determined in serum by RRA and expressed in cis(z)-flupenthixol equivalents. The drug level was significantly correlated to the dose. No clear relationship between drug level and clinical results as well as side effects was found. Less pronounced variations of the drug level between subsequent injections resulted in a positive therapeutic response.     

Determination of small quantities of sulfate (0-12 nmol) in serum, urine, and cartilage of the mouse

The colorimetric benzidine method of K. S. Dodgson and B. Spencer (1953, Biochem. J. 55, 436-440) for the measurement of inorganic sulfate can be scaled down about 100 times by using disposable 96-well microplates instead of individual cuvettes. Ten-microliter samples of serum and urine, derived from mice, can be analyzed in a simple, rapid, and reliable way without sacrificing the animals. Without prior isolation of sulfated glycosaminoglycans, ester sulfate in mouse patellar cartilage is liberated quantitatively as inorganic sulfate upon acid hydrolysis in 3 M HCl for 16 h at 80 degrees C. To this end the articular cartilage layer of the patella must be separated in toto from the underlying bone. Subsequent hydrolysis in polypropylene tubes gives accurate results. In contrast, hydrolysis in borosilicate glass vials is useless, since nanomoles of sulfate added cannot be recovered adequately. The thin patellar cartilage layer obtained from 10-week-old male mice contains about 5 nmol of sulfate, an amount easily measured with the developed microplate benzidine method.     

An nptI-sacB-sacR cartridge for constructing directed, unmarked mutations in gram-negative bacteria by marker exchange-eviction mutagenesis

A technique for marker exchange-eviction mutagenesis that enables the construction of directed, unmarked mutations in Gram-negative bacteria was demonstrated in Erwinia chrysanthemi. The technique employs an nptI-sacB-sacR cartridge that is carried on a 3.8-kb BamHI fragment and confers kanamycin (Km) resistance and sucrose sensitivity (due to the production of levansucrase by sacB) in E. chrysanthemi. The cartridge was inserted into a Sau3A site in a cloned E. chrysanthemi pelC gene (encoding pectate lyase isozyme PLc) and then introduced into the Erwinia genome by gene exchange recombination. The resulting mutant was KmR, sucrose-sensitive, and PLc-deficient. The cartridge was then excised from the plasmid-borne pelC gene by PstI cleavage to leave a 28-bp frame-shifting insertion. The pelC allele containing the 28-bp insertion was exchanged for the chromosomal allele containing the nptI-sacB-sacR cartridge by selection for sucrose tolerance. The resulting E. chrysanthemi mutant was Kms and PLc-deficient. The technique permits the construction of complex strains with many directed mutations without the introduction of a corresponding number of antibiotic resistance markers and should prove useful, for example, in exploring the role of the multiple pel genes in E. chrysanthemi.     

Simulation of the effects of Acanthaster planci on the population structure of massive corals in the genus Porites: evidence of population resilience?

Scleractinian corals in the genus Porites are slow growing, can live for centuries, and can attain great size. In these respects they differ from the majority of coral species, which grow faster and live for years to decades. The predatory starfish Acanthaster planci L. feeds on a wide range of coral species including Porites spp., and during outbreaks in its populations, causes high coral mortality and injury over much of the affected reefs. Because they are slow growing and because recent outbreaks of the starfish occurred only 15 years apart, it may be argued that the Porites populations on affected reefs will be sent into a period of prolonged decline. The present study uses a size stage model of the transition matrix type to predict effects of starfish outbreaks of various frequencies on Porites populations. A transition matrix characterizing the mortality and injury caused in different Porites size classes at John Brewer Reef during an outbreak year was determined from field data. Transition matrices for non-outbreak years were constructed on the basis of realistic growth rates and postulated survivorship and recruitment schedules. The medium term (100 years) effects of outbreaks were simulated by alternation of a single iteration of the outbreak matrix with many iterations of each non-outbreak matrix. By varying the interval between simulated outbreaks it was possible to define combinations of growth rate, survivorship and recruitment which were viable for various outbreak intervals. Simulations based on estimates of the initial size frequency distribution, recruitment rates and colony growth rates for the John Brewer Reef population predicted that the population would remain viable in the face of outbreaks every 15 years only if juvenile and adult survivorship were high. However, within the range of colony growth rates known to occur throughout the Great Barrier Reef and at recruitment rates of the same order as those estimated in the field population, it appears that a much wider range of survivorship schedules could lead to parity or even sustained growth in the face of outbreaks recurring at intervals of from 1 to 3 decades. It is suggested that because the key measurable parameters (initial size structure, damage characteristics, recruitment rate and growth rate) are likely to be very patchy at the scale of whole reefs, no general statement concerning the prognosis for Porites would be meaningful. However the model provides a tool by which a standardized evaluation of this type of field data may be made on a reef by reef basis.     

Modification of root bud growth in Canada thistle with selected plant growth regulators: effects on translocation of glyphosate

Root applications of 0.1 M 6-benzyladenine (BA) and 10.0 M indole-3-butyric acid (IBA) enhanced or inhibited, respectively, root bud growth in hydroponically grown Canada thistle [Cirsium arvense (L.) Scop.]. Translocation of¹⁴C-glyphosate [N-(phosphonomethyl)glycine] into roots was positively correlated with this growth. Foliar applications of ethephon or chlorfluorenol also enhanced root bud growth, but glyphosate translocation was only weakly correlated with such growth in soil-grown Canada thistle. At glyphosate rates above 0.56 kg/ha, root bud growth was not stimulated by plant growth regulators (PGRs) and basipetal translocation was not enhanced. Paradoxically, ethephon and chlorflurenol restrained root bud growth in the field since thistle control steadily improved during the 3 years following treatment.     

Phytohormone effects on cell division inChlorella pyrenoidosa chick (TX-7-11-05) (chlorellaceae)

Phytohormone effects on cell division of synchronous cultures ofChlorella pyrenoidosa (TX-7-11-05) were studied under different photo flux densities. The time required for initiation of incipient cell division was reduced significantly by treatment with kinetin (6-furfurylaminopurine), gibberellic acid, and indole-3-acetic acid. Data show that kinetin was more effective than gibberellic acid, which was more effective than indole-3-acetic acid. Studies with combinations of the phytohormones revealed no antagonistic, additive, or synergistic effects.