Amplified fragment length polymorphism analysis of the VNTR locus D1S80 in central Spain

Summary The polymorphism of the D1S80 locus has been analyzed in a population sample of 203 unrelated individuals living in Madrid (central Spain) by PCR and subsequent semi-dry discontinuous polyacrylamide gel electrophoresis (Tris-chloride/Tris-glycine buffer system) followed by silver staining, The electrophoretic system described in this study offers high resolution in the separation of the different D1S80 alleles allowing the detection of microvariability around the allele T22 in the spanish population. Twenty different alleles containing 17–40 repeats of the basic 16bp unit were distinguished. The alleles T18 and T24 were found to be relatively common in Spain, as in other populations:, with frequencies of 0.224 and 0.372, respectively. No evidence of significant deviations from Hardy-Weinberg equilibrium was found in these preliminary population data.     

The applicability and utility of the MMPI-based offender classification system in a correctional mental health unit

This study investigated the applicability and utility of Megargee and Bohn's MMPI-based offender classification system in correctional mental health units (MHUs). Previous studies found that 11 MHU samples (n = 1723) had substantially more offenders classified in the more pathological MMPI types than did 21 samples (n = 5881) drawn from general male populations in US prisons. In this study of 63 severely disturbed felons, 43% belonged to the most pathological type (‘group How’). Comparing MHU patients with general offenders from the same IvfIvIPI types on staff ratings and case history variables, we found that the MHU patients were significantly poorer in adjustment. Within the MHU sample, there was no difference in case history variables or adjustment ratings between those in the most and least severe MMPI types. These findings differed from those of studies using less severely disturbed, more heterogeneous, MHU populations. It was concluded that, in settings in which the entire population is flagrantly disturbed, the MMPI-based system is more useful in screening potential admissions than it is in making meaningful distinctions among those already admitted.     

ALFENTANIL PLASMA CONCENTRATION v. EFFECT RELATIONSHIPS IN CARDIAC SURGICAL PATIENTS

Effects of alfentanil, preceded by lorazepam, on suppressionof haemodynamic and somatic responses to noxious stimuli wasstudied in patients undergoing CABG. Plasma concentration ofalfentanil, somatic and haemodynamic responses were measuredat loss of consciousness, tracheal intubation, sternotomy andduring multiple applications of electrocoagulation. Additionalalfentanil was administered i.v. to control unwanted responses.Study 1 (six patients): lorazepam 0.08 mg kg^–1 by mouth1–2 h before operation, alfentanil priming infusion (60µg kg^–1 min^–1 for 10 min) followed by maintenanceinfusion (4.5 µg kg^–1 min^–1). With mean plasmaalfentanil 1178 (SEM 54) ng ml^–1, two patients requiredsupplementary alfentanil to suppress somatic motor responses;one patient required nitroglycerin to control an increase inarterial pressure which was unresponsive to additional alfentanilfollowing sternotomy. Study 2 (13 patients): lorazepam 0.04mg kg^–1 by mouth as premedication; one of three maintenanceinfusion rates of alfentanil: 5.4 (n=4), 6.6 (n=5), or 7.8 (n=4)µg kg^–1 min^–1, each preceded by a proportionalpriming infusion. With plasma alfentanil 2181 (62)ng ml^–1,somatic motor responses requiring additional alfentanil occurredin nine patients; haemodynamic responses in four of seven patientstested could not be controlled by alfentanil. The highest plasmaconcentration of alfentanil to prevent response to a stimulusother than tracheal intubation was different between the twostudies (P<0.05). We conclude that alfentanil alone is insufficientto suppress haemodynamic and somatic motor responses to noxiousstimulation during CABG and that the role of premedication issignificant. *Department of Anesthesia, Bowman-Gray School of Medicine Winston-Salem,NC 27103, U.S.A. 2114 de Mayo Road, Del Mar, Ca. 92014, U.S.A.     

Enzymatic synthesis of penicillins

Different penicillins (phenylacetyl, 2-hydroxyphenylacetyl, 4-hydroxyphenylacetyl, phenoxyacetyl and 2-thiopheneacetylpenicillin) have been synthesized "in vitro" by direct N-acylation of 6-aminopenicillanic acid (6-APA) with the acyl group of several acyl-CoA derivatives. The enzyme that catalyzes these reactions, acyl-CoA: 6-APA acyltransferase of Penicillium chrysogenum, was purified to homogeneity (374-fold) and its amino acid composition is given. This protein accepts as substrates several aliphatic acids and different aromatic acids with the only requirement that an acetyl-CoA moiety must be present in the substrate molecule. Shortening or lengthening of the acyl moiety prevents the 6-APA-N-acylation reaction. The presence of an amino group in the alpha-position of the acetyl group does not allow this molecule to be used as substrate. However, different substitutions in the phenyl group (hydroxylation of the carbons 2 and 4) or its replacement by another aromatic ring (thiophene) were accepted with varying reactions rates in the acylation reaction when a 176-fold purified acyltransferase was employed. The homogeneity pure enzyme accepts as substrate thiophene acetyl-CoA but it did not 2-hydroxyphenyl and 4-hydroxyphenylacetyl-CoA. The presence of an oxygen atom between the aromatic and the acetyl moieties did not affect the catalysis.     

An Immunotoxicological Evaluation of 4, 4'-Thiobis-(6-t-butyl-m-cresol) in Female B6C3F1 Mice: 1. Body and Organ Weights, Hematology, Serum Chemistries, Bone Marrow Cellularity, and Hepatic Microsomal Parameters

An Immunotoxicological Evaluation of 4,4'-Thiobis-(6-t-butyl-m77-cresol)in Female B6C3F1 Mice. 1. Body and Organ Weights, Hematology,Serum Chemistries, Bone Marrow Cellularity, and Hepatic MicrosomalParameters. MUNSON, A. E., WHITE, K. L., JR., BARNES, D. W.,MUS-GROVE, D. L., LYSY, H. H., AND HOLSAPPLE, M. P. (1988).Fundam. Appl. Toxicol. 10, 691–700. Adult female B6C3F1mice were gavaged with 4,4'-thiobis-(6-t-butyl-m-cresol) (TBBC)in corn oil at doses of 10, 100, or 200 mg/kg daily for 14 consecutivedays. There was no overt toxicity, as manifested by grosslyobservable behavioral changes, decreased growth rate over theexposure period, or mortality. There were also no marked effectson serum chemistries or hematology, with the exception of asignificant increase (41%) in the number of leukocytes at thehighest dose. Absolute differential counts indicated that significantincreases occurred in the number of lymphocytes (31%) and neutrophils(177%). Studies with bone marrow indicated a significant 30%increase in the number of cells/femur from animals treated withthe highest dose of TBBC. The number of macrophage progenitors(CFU-M)/femur was significantly increased by 28%, while thenumber of granulocyte-monocyte progenitors (CFU-GM)/femur wasnonsig-nificantly increased by 20% in the high dose animals.The weight of both the spleen and liver was increased in a dose-relatedfashion, although the histopathology of the spleen of TBBC-treatedmice was not different from control. The livers of mice receivingthe high dose showed mild focal hydropic degeneration, mildhepatitis, and a slight increase in the number of Kupffer cells.No other organs were affected. Liver microsomal protein andcytochrome P-450 levels were increased in a dose-related fashion.Enzyme activities of aminopyrine demethylase and aniline hydroxylase,but not arylhydrocarbon hydroxylase, were also increased ina dose-related fashion.     

Immunotoxicity of the Semiconductor Gallium Arsenide in Female B6C3F1 Mice

The effects of gallium arsenide (GaAs) exposure on immunocompetenceof B6C3F1 female mice were investigated. GaAs was administeredas a single intratracheal instillation at doses of 50, 100,and 200 mg/kg. Fourteen days after exposure, various cellularand humoral immune parameters were assessed. GaAs exposure increasedspleen cellularity in a dose-dependent manner. However, thepercentages of Thy 1.2 positive and 1g positive cells were decreasedand that of F4/80 positive cells was increased dose dependency.The IgM and IgG antibody-forming cell response of the spleento the T-dependent antigen sheep erythrocytes was reduced by66 and 48%, respectively, at 200 mg/kg. Levels of the serumcomplement protein, C3, were increased by as much as 16% withno significant change in CH50 levels. The mitogenic responseof splenic T cells to Con A and PHA was unaffected by GaAs,but that of B cells to LPS was increased by 52%. The delayedhypersensitivity response to keyhole limpet hemocyanin and mixedlymphocyte response were significantly reduced in a dose-dependentmanner by GaAs exposure. Natural killer cell activity againstthe YAC-1 mouse lymphoma was enhanced in treated mice. Analysisof peritoneal exudate cells (PEC) revealed a dose-dependentdecrease in number and a shift in the composition of PECs. Thepercentage of PEC monocytes increased from 53% of the populationto 81%, while the lymphocytes decreased from 46 to 20%. Theadherent PEC population demonstrated decreased phagocytosisof covaspheres and increased phagocytosis of chicken erythrocytes(CRBC). GaAs exposure had no effect on host resistance to Plasmodiumyoelii or Streptococcus pneumoniae, but dose dependency increasedresistance of the mouse to Listeria monocytogenes Treated micedemonstrated a significantly decreased resistance to the B16F10melanoma with a sevenfold increase in tumor burden at 200 mg/kg.GaAs affects both humoral and cellular immune parameters inmice and impairs the ability of the immune system to protectagainst B16F10 tumor challenge.     

Spike patterning by Ca2+-dependent regulation of a muscarinic cation current in entorhinal cortex layer II neurons

In entorhinal cortex layer II neurons, muscarinic receptor activation promotes depolarization via activation of a nonspecific cation current (I(NCM)). Under muscarinic influence, these neurons also develop changes in excitability that result in activity-dependent induction of delayed firing and bursting activity. To identify the membrane processes underlying these phenomena, we examined whether I(NCM) may undergo activity-dependent regulation. Our voltage-clamp experiments revealed that appropriate depolarizing protocols increased the basal level of inward current activated during muscarinic stimulation and suggested that this effect was due to I(NCM) upregulation. In the presence of low buffering for intracellular Ca(2+), this upregulation was transient, and its decay could be followed by a phase of I(NCM) downregulation. Both up- and downregulation were elicited by depolarizing stimuli able to activate voltage-gated Ca(2+) channels (VGCC); both were sensitive to increasing concentrations of intracellular Ca(2+)-chelating agents with downregulation being abolished at lower Ca(2+)-buffering capacities; both were reduced or suppressed by VGCC block or in the absence of extracellular Ca(2+). These data indicate that relatively small increases in [Ca(2+)](i) driven by firing activity can induce upregulation of a basal muscarinic depolarizing-current level, whereas more pronounced [Ca(2+)](i) elevations can result in I(NCM) downregulation. We propose that the interaction of activity-dependent positive and negative feedback mechanisms on I(NCM) allows entorhinal cortex layer II neurons to exhibit emergent properties, such as delayed firing and enhanced or suppressed responses to repeated stimuli, that may be of importance in the memory functions of the temporal lobe and in the pathophysiology of epilepsy.