The history of the cattle tick Boophilus microptus in Australia and achievements in its control

For more than 100 years, tick fever and the cattle tick have caused tremendous financial loss to cattle producers around the world. Since Australia became infected with the disease and infested with its tick vector in the mid-19th century, a great deal of research effort has been directed towards their effective control by Australian farmers, administrators and scientists. Such research has yielded information which has facilitated the development of various control strategies that have equal application in other countries afficted with the same problem. It has been demonstrated that integration of a variety of these strategies is necessary for long-lasting control.     

Catecholamines in fetal pig plasma and the response to acute hypoxia and chronic fetal decapitation

Summary Dopamine, norepinephrine and epinephrine were measured by radioenzymatic assay in blood plasma samples drawn from the umbilical arteries of 30 anaesthetised Landrace pig fetuses. Just prior to term, the concentrations of dopamine (0.46±0.14 ng·ml^–1) and norepinephrine (1.74±0.60 ng·mg^–1) were lower than earlier in gestation, whereas epinephrine concentrations at term (0.80±0.31 ng·ml^–1) were similar to those at mid-gestation, intervening stages of gestation having higher levels of plasma epinephrine. Fetal hypoxia was induced by clamping the umbilical cord for 2 min and the catecholamines determined in arterial blood samples immediately thereafter, then again 3 min after removal of the clamp. Inconsistent effects of cord clamping on catecholamine levels were seen at 55 days, but thereafter, in all but one instance, the hormone levels were increased. Fetuses near term tended to respond less than fetuses at 75 and 96 days gestation (term=114±1 day). Catecholamines were also present in the circulation of fetuses decapitated at 42 days gestation and studied at 109±1 days. The average concentrations of dopamine (1.12±0.27 ng·ml^–1) and norepinephrine (8.23±3.04 ng·ml^–1) were greater than in intact fetuses, the plasma epinephrine levels being comparable to, or slightly higher than, those in intact fetuses. The results demonstrate that catecholamines are present in the circulation of the intact and decapitated pig fetus and that the actual concentrations and the type of response to umbilical cord clamping are dependent on gestation age.     

Metabolite and ionic composition of follicular fluid from different-sized follicles and their relationship to serum concentrations in dairy cows

Metabolic changes in blood serum may be reflected in the biochemical composition of follicular fluid and could indirectly influence oocyte quality. The purpose of this study was to examine the biochemical composition of follicular fluid harvested from different-sized follicles and its relationship with that of blood serum in dairy cattle. Following slaughter, blood samples were collected from dairy cows (n=30) and follicular fluid aspirated from three size classes of non-atretic follicles (<4 mm, 6–8 mm and >10 mm diameter). Samples remained independent between cows and between size classes within cows. Serum and follicular fluid samples were assayed using commercial clinical and photometric chemistry assays for ions (sodium, potassium and chloride) and metabolites (glucose, β-hydroxybutyrate (β-OHB), lactate, urea, total protein, triglycerides, non-esterified fatty acids (NEFA) and total cholesterol). Results showed that follicular fluid concentrations of glucose, β-OHB and total cholesterol increased from small to large follicles and decreased for potassium, chloride, lactate, urea and triglycerides. There was a significant concentration gradient for all variables between their levels in serum and follicular fluid (P<0.05). Significant correlations were observed for chloride (r=0.40), glucose (r=0.56), β-OHB (r=0.85), urea (r=0.95) and total protein (r=0.60) for all three follicle size classes and for triglycerides (r=0.43), NEFA (r=0.50) and total cholesterol (r=0.42) for large follicles (P<0.05). The results from the present study suggest that the oocyte and the granulosa cells of dairy cows grow and mature in a biochemical environment that changes from small to large follicles. Furthermore, the significant correlation between the composition of serum and follicular fluid for the above-mentioned metabolites suggests that metabolic changes in serum levels will be reflected in the follicular fluid and, therefore, may affect the quality of both the oocyte and the granulosa cells.     

Strategies for rare-event detection: an approach for automated fetal cell detection in maternal blood.

This article explores the feasibility of the use of automated microscopy and image analysis to detect the presence of rare fetal nucleated red blood cells (NRBCs) circulating in maternal blood. The rationales for enrichment and for automated image analysis for "rare-event" detection are reviewed. We also describe the application of automated image analysis to 42 maternal blood samples, using a protocol consisting of one-step enrichment followed by immunocytochemical staining for fetal hemoglobin (HbF) and FISH for X- and Y-chromosomal sequences. Automated image analysis consisted of multimode microscopy and subsequent visual evaluation of image memories containing the selected objects. The FISH results were compared with the results of conventional karyotyping of the chorionic villi. By use of manual screening, 43% of the slides were found to be positive (>=1 NRBC), with a mean number of 11 NRBCs (range 1-40). By automated microscopy, 52% were positive, with on average 17 NRBCs (range 1-111). There was a good correlation between both manual and automated screening, but the NRBC yield from automated image analysis was found to be superior to that from manual screening (P=.0443), particularly when the NRBC count was >15. Seven (64%) of 11 XY fetuses were correctly diagnosed by FISH analysis of automatically detected cells, and all discrepancies were restricted to the lower cell-count range. We believe that automated microscopy and image analysis reduce the screening workload, are more sensitive than manual evaluation, and can be used to detect rare HbF-containing NRBCs in maternal blood.     

Various synaptic activities and firing patterns in cortico-striatal and striatal neurons in vivo

It is commonly assumed that spontaneous activity of striatal output neurons is characterized by a two-state behavior. This assumption is mainly based on in vivo intracellular recordings under urethane and/or ketamine-xylazine anesthesia showing that striatal neurons oscillate between two preferred membrane potentials, a Down state (hyperpolarized level), resulting from an inwardly rectifying potassium conductance, and an Up state (depolarized level) caused by complex interactions between a barrage of cortical synaptic excitation and voltage-dependent potassium conductances. However, a recent comparative study using different anesthetics showed that striatal neurons can exhibit various shapes of synaptic activity depending on the temporal structure and the degree of synchronization of their cortico-striatal afferents. These new data demonstrate that the "classical" Up and Down states do not provide the unique spontaneous activity that can be encountered in striatal neurons in vivo. Rather we propose that striatal neurons should exhibit various synaptic activities and firing patterns depending on the states of vigilance. This hypothesis would be validated in further experiments in which the intracellular activity of striatal neurons will be recorded during the natural sleep-wake cycle.     

A new approach to reaching control for tetraplegic subjects

A simple upper limb control strategy to guide reaching in preparation to grasp for tetraplegic subjects is proposed. The control is based on new studies of self-paced human arm movements involving rotations about the shoulder and elbow joints. An experimental study of reaching, while grasping, by able-bodied humans, allowed us to reduce the dimensionality of the control vector from two to a single variable. This was accomplished by detailed analysis of the synergy between shoulder and elbow joint angles. This study examined only movements in teh horizontal plane. In the experiments we varied: (a) the shape of targets; (b) their position relative to the initial position of the hand; and (c) the speed of reaching. A synergy between shoulder and elbow joint angles was found in most analysed movements, and it was characterized by a scaling parameter between elbow and shoulder angular velocities. The scaling parameter was determined from the target position presented in the visual perceptive field and initial shoulder and elbow angles. The same experimental setup in studies with tetraplegics with retained shoulder movements showed that this natural synergism is preserved even though the motor and sensory components of the upper limb are reduced or absent. Tetraplegics originally showed a very different reaching pattern, but after short training sessions they developed a reaching behaviour which was similar to able-bodied subjects. The results presented can be used in the following way: a tetraplegic subject lacking elbow extension and flexion may be fitted with an assistive system which will be volitionally controlled only from ipsilateral shoulder movements. The assistive system can comprise either a motorized brace, or a functional electrical stimulation system applied to elbow flexors and extensors. With this system volitional movements at the shoulder would bring the hand into the correct position to accomplish an assisted grasping motion.     

A method of thymectomy in macaques

The aim of this study was to develop an operation for total thymectomy suitable for nonhuman primates. The thymus was removed from 24 macaque monkeys from fetal to adult life by a cervicothoracotomy and blunt gauze dissection. Mean wet weight of the thymus rose from 150 mg (range 50-250 mg) in fetal animals to 2.5 gm (range 0.3-4.2 gm) in prepubertal monkeys. Nonhuman primate thymic weight fell steadily and uniformly with age by over 300% when plotted as a function of body weight. This surgical technique appears to be a valid method of total thymectomy in macaque monkeys.     

A method for assessing the equivalence of repeated measures of muscle fatigue rates estimated from EMG power spectrum analysis

This report addresses some of the statistical problems that are encountered when the test/retest recording reliability of fatigue-related parameters of the EMG power spectrum is evaluated. It can be shown that some classical methods for reliability assessment such as correlational procedures are unsuitable for this purpose. Because the EMG power spectrum fatigue parameter depends on metabolic changes in muscle tissue, it is suggested that methods similar to those used in the evaluation of bioequivalence studies may be more appropriate in the assessment of such test/retest results.     

Variability of the EMG mean power frequency: A study on the trapezius muscle

Calculation of the EMG mean power frequency (MPF) is a common procedure applied in evaluation of the frequency shift associated with local muscle fatigue. Variations of the MPF that are unrelated to muscle fatigue may jeopardize the estimation of the frequency shift. Different kinds of variation include random variation and systematic variation due to changes in posture or load. In a previous article we have evaluated the systematic linear variation of the MPF. The aim of the present study was to examine the random variation. Data sequences of 10 s, each obtained from nonfatigued trapezius muscle of 19 healthy subjects, were examined over a functional range of load and joint angles with multiple regression analysis. The random variation was evaluated with residual analysis. The residual standard deviation within the whole group was 10% for surface recordings and 13% for intramuscular recordings. If only within-subject variation was considered, the corresponding values were 5 and 8%. Based on this, confidence and prediction intervals for the regression models were calculated. Ninety-five percent confidence intervals were ±1–3% around the regression surfaces, whereas 95% prediction intervals for single measurements were as large as ±20–26% for the whole group, and ±11–20% if only within-subject variations were considered. Assessment of localized muscle fatigue using single MPF estimates should therefore be avoided. Multiple measurements and regression analysis are discussed as methods to minimize the effects of random variations.     

Tectal responses to potassium loads and subsequent visual stimuli in the toad, Bufo bufo

Calling male toads were tested behaviourally for their prey catching responses to wormlike stimuli and assigned to groups of non-hungry and hungry depending on their prey catching motivation before being prepared for visual unit, massed unit and slow potential shift (SPS) recording from the optic tectum. Control recordings to visual stimuli were made before recording the effects of application of isotonic solutions containing concentrations of 0-41 mM K(+). Application of solution was followed by presentation of the visual stimulus while the solution still bathed the tectum. The best tectal responses were made to large square visual stimuli in the non-hungry toads, perhaps because recordings were made in the breeding season. Responses of the tectum to solution addition were significant in the concentration range of 7-17 mM K(+). Hungry toads showed an earlier, smaller response than non-hungry (sexually motivated) animals. When the visual stimulus was presented, there were unit and massed unit responses at all bathing solution concentrations, which were larger in non-hungry animals. These experiments revealed that toads motivated to feed respond earlier than non-hungry toads to application of artificial CSF to the tectum, though non-hungry toads responded best to the subsequent visual stimulus.     

A method facilitating the demonstration of synergism in the tissue-responses to mixed peptide stimuli

A methodology for the pharmacodynamical assay of mixtures of 2 spasmogenic peptides is described, illustrated and discussed, with reference to a method intended for the chemists (P. Job, 1927). This allows a quick appreciation of the existence of synergism, of its own order of magnitude and of that of the doses for which it is significant. Also given is a fast way of curve-fitting the dose response to individual peptides, according to Clark's approximation.     

异丙酚用于腰硬联合麻醉对患者围术期氧代谢的影响

目的:探讨异丙酚复合腰硬联合麻醉对腹腔镜手术患者氧代谢的影响。方法:选取2012年5月-2013年7月在我院进行妇科腹腔镜手术的患者153例,均采用异丙酚复合腰硬联合麻醉。观察并记录患者在麻醉前、麻醉后、气腹15 min、30 min、60 min及放气腹15 min的氧代谢。结果;与麻醉前比较,麻醉后及气腹过程中的氧供(DO2)较麻醉前下降,但无统计学意义(P0.05);而氧耗(VO2)、摄氧率(ERO2)均明显下降,差异具有统计学意义(P0.05);pH值、[HCO3-1]、动脉二氧化碳分压(PaCO2)、混合静脉二氧化碳分压(PvCO2)、动脉氧分压(PaO2)、混合静脉氧分压(PvO2)、动脉血氧浓度(SaO2)及混合静脉血氧浓度(SvO2)与均呈显著性差异,具有统计学意义(P0.05);放气腹后15 min各项参数与麻醉前比较,差异无统计学意义(P0.05)。结论:异丙酚复合腰硬联合麻醉效果良好,但对患者的氧代谢有一定影响,手术中应注意检测患者的呼吸循环功能。     

Itraconazole and leishmaniasis: A randomised double-blind trial in cutaneous disease

Cutaneous leishmaniasis (CL) is a vector-borne parasitic disease of the skin. Previous open controlled studies with oral itraconazole suggest that it was effective for CL in India. Twenty patients with localised CL participated in this trial. Patients were allocated randomly to receive capsule itraconazole and matching placebo for 6 weeks. No topical medicines were used. Demonstration of Leishmania by slit smear was mandatory. Prior to, periodically during and 3 months after completion of therapy an overall clinical assessment, liver function tests and urinalysis were performed. On decoding, out of the 10 cases receiving drug itraconazole, 7 were declared cured by clinical and parasitological criteria. No major side-effects were noted. Spontaneous remission was observed in 1 case in the placebo group at 3 months follow up. Oral itraconazole has a promising antileishmanial potential and may thus secure CL patient from the hazards of antimonials.     

A general procedure for the purification of human beta-secretase expressed in Escherichia coli

Expression and purification of human beta-secretase (BACE1) in bacteria have been plagued with issues concerning solubility, inhomogeneous N-terminus, and lack of enzymic activity. Several forms of the mature human BACE1 have been expressed in Escherichia coli with different N-terminal extensions and without the C-terminus transmembrane domain. Although each of the proteins expresses in inclusion bodies, a generalized protocol has been developed to solubilize, refold, and purify these BACE1 variants. The resultant proteins are homogeneous and monodispersed in solution. Each possesses a unique N-terminus. Activity assays using the peptide substrate 7-methoxycoumarin-4-yl-SEVNLDAEFK-2,4-dinitrophenyl-RR, corresponding to the beta-secretase cleavage sequence in the amyloid precursor protein with the Swedish mutations of N(670)L(671) substituting for the residues K(670)M(671), reveal a kcat and KM of 9.3 min(-1) and 55 microM, respectively.     

Potential for interactions between the carboxy- and amino-termini of Rubisco activase subunits

The subunit interactions of Rubisco activase were investigated using mutants containing an introduced Cys near the N- and/or C-terminus. Chemical cross-linking of the C-terminal and double insertion mutant produced subunit dimers and dimers plus high ordered oligomers, respectively. Fluorescence measurements with N,N′-dimethyl-N-(iodoacetyl)-N′-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)ethylenediamine showed that the environment around the introduced Cys near the C-terminus becomes more hydrophilic upon nucleotide binding. The Cys insertion mutants catalyzed Rubisco activation and ATP hydrolysis even when the subunits of the C-terminal or double insertion mutants were completely cross-linked. The results indicate that the termini of adjacent activase subunits are in close proximity and can be modified and even joined without affecting enzyme function.     

Observations on the history of Dutch physical stature from the late-Middle Ages to the present

In the late-Middle Ages and at the onset of the early modern period, the Dutch population was taller than in the first half of the 19th century. This inference is partially based on skeletal evidence, mainly collected by the Dutch physical anthropologist George Maat and his co-workers. A spectacular increase in Dutch heights began in the second half of the 19th century and accelerated in the second half of the 20th century. At the end of the 20th century, the Dutch became tallest in the world.     

Description of trophic status, hyperautotrophy and dystrophy of a coastal lagoon through a potential oxygen production and consumption index—TOSI: Trophic Oxygen Status Index

Seasonal changes of trophic status of the Sacca di Goro lagoon (northern Italy) are analysed through time series of oxygen metabolism for two stations in which separate measures of planktonic, macroalgal (Ulva rigida), and benthic exchange are available. These component results and their sums (i.e. total ecosystem metabolism) are compared using conventional analyses and an index obtained from oxygen fluxes (Trophic Oxygen Status Index, TOSI). The TOSI is derived from the Benthic Trophic Status Index (BTSI) proposed earlier by Rizzo et al. [Estuaries 19 (1996) 247] and basically represents the net potential metabolism. The index results from the relationship between net maximum productivity (NP), measured at saturating light, and dark respiration (DR). The index was developed to provide a simple portrayal of oxygen processing over time and space for shallow aquatic systems and has two modes: a categorical classification and a graphical representation. The categorical classification of the index from autotrophy to heterotropy provides a rapid assessment of the potential oxygen balance and thus evidences critical situations in the lagoonal metabolism. In the graphical representation of the TOSI, three pieces of information are given: the categorical TOSI, the magnitude of flux for both NP and DR and the time line of the fluxes. Where flushing is slow, the TOSI reflects dissolved oxygen dynamics since the NP:|DR| ratio clearly correlates with both the maximum oxygen concentration (MOC) and the daily quantity of oxygen remaining in the water column (RO). However, TOSI is less well related to MOC and RO in open systems in which oxygen concentrations are dominated by physical factors. The graphical representation of TOSI seems also suitable to represent the degree of intrasystem disturbance that is related to the excess of primary production and changes in oxygen availability. It can also discriminate among different photoautotrophic conditions, including hyperautotrophy, as an abnormal oxygen production with respect to the biomass build up, and dystrophy, as the subsequent abnormal oxygen deficit which causes prolonged anoxia and the onset of anaerobic metabolism. Overall, the index provides a tool for rapid assessment of system metabolism and potentially its consequences.     

Haplotype mapping of a major quantitative-trait locus for fetal hemoglobin production, on chromosome 6q23.

Fetal hemoglobin (Hb F) and fetal cell (FC) levels in adults show considerable variation and are influenced by several genetic variants; the major determinants appear to be unlinked to the beta-globin gene cluster. Recently, a trans-acting locus controlling Hb F and FC production has been mapped to chromosome 6q23 in an Asian Indian kindred that includes individuals with heterocellular hereditary persistence of Hb F (HPFH) associated with beta thalassemia. We have extended the kindred by 57 members, bringing the total studied to 210, and have saturated the region with 26 additional markers. Linkage analysis showed tight linkage of the quantitative-trait locus (QTL) to the anonymous markers D6S976 (LOD score 11.3; recombination fraction .00) and D6S270 (LOD score 7.4; recombination fraction .00). Key recombination events now place this QTL within a 1-2-cM interval spanning approximately 1.5 Mb between D6S270 and D6S1626. Furthermore, haplotype analysis has led to a reevaluation of the genealogy and to the identification of additional relationships in the kindred.     

Expression of ricin A chain and ricin A chain-KDEL in Escherichia coli

Ricin and its A chains can be used to conjugate with monoclonal antibodies to prepare immunotoxins. Ricin A chain (RTA) and its modification RTA-KDEL (ER-retrieval signal) were expressed with the pKK223.3 system in Escherichia coli under control of a tac promoter. The recombinant proteins can be purified by one-step affinity chromatography on a column of Blue-Sepharose 6B. The toxicities of RTA and its mutant RTA-KDEL were evaluated by the MTT assay in HeLa, MCF, and ECV-304 cells following fluid-phase endocytosis. RTA-KDEL was somewhat more cytotoxic than RTA itself in the different cell lines. The results suggest that rRTA-KDEL may be useful for the synthesis of more potent immunotoxins.     

Prediction of amino acid pairs sensitive to mutations in the spike protein from SARS related coronavirus

In this study, we analyzed the amino acid pairs affected by mutations in two spike proteins from human coronavirus strains 229E and OC43 by means of random analysis in order to gain some insight into the possible mutations in the spike protein from SARS-CoV. The results demonstrate that the randomly unpredictable amino acid pairs are more sensitive to the mutations. The larger is the difference between actual and predicted frequencies, the higher is the chance of mutation occurring. The effect induced by mutations is to reduce the difference between actual and predicted frequencies. The amino acid pairs whose actual frequencies are larger than their predicted frequencies are more likely to be targeted by mutations, whereas the amino acid pairs whose actual frequencies are smaller than their predicted frequencies are more likely to be formed after mutations. These findings are identical to our several recent studies, i.e. the mutations represent a process of degeneration inducing human diseases.