Freezing point and other technological properties of milk of the Polish Coldblood horse breed.

Mare milk is a valued and sought-after raw material for the production of innovative dairy products. The high demand, low supply, high price, and lack of accurate characterization of the milk of a given horse breed may provoke its deliberate fraudulent dilution. The aim of this work was to analyze the freezing point against a background of various selected technological parameters of Polish Coldblood mare milk. Research was carried out on multiparous Polish Coldblood mares from 5 to 8 yr of age with live weights between 618 and 851 kg. Their milk was tested on d 1, 3, and 7 postpartum and once a month for the next 9 mo of lactation (n = 52). Milk from 13 mares, taken daily in mo 4 of lactation, was collected for the dilution study (n = 403). The basic composition of the milk was as follows: solids-not-fat, 84.4 g/kg, fat 15.1 g/kg, lactose 65.3 g/kg, and total protein 24.2 g/kg (made up of casein, 14.6 g/kg, and whey protein, 9.5 g/kg). The dominant amino acids were glutamic acid, proline, leucine, and aspartic acid. The mean freezing point during a 9-mo lactation was -0.557°C (coefficient of variation = -2.787; 5th to 95th percentile: -0.563 to -0.550°C). Based on our analysis, we adopted -0.550°C (from 3 to 6 mo) as the reference value for milk from the Polish Coldblood horse. Analysis of the model of changes in freezing point, after dilution, using the coefficient of determination and Akaike's information criterion, showed that a linear model could be recommended (y = 79.35 + 144.35x). Addition of 1% water increased the freezing point of Polish Coldblood mare milk by 0.0069°C.

Excitation and electron transfer in reaction centers from Rhodobacter sphaeroides probed and analyzed globally in the 1-nanosecond temporal window from 330 to 700 nm.

Global analysis of a set of room temperature transient absorption spectra of Rhodobacter sphaeroides reaction centers, recorded in wide temporal and spectral ranges and triggered by femtosecond excitation of accessory bacteriochlorophylls at 800 nm, is presented. The data give a comprehensive review of all spectral dynamics features in the visible and near UV, from 330 to 700 nm, related to the primary events in the Rb. sphaeroides reaction center: excitation energy transfer from the accessory bacteriochlorophylls (B) to the primary donor (P), primary charge separation between the primary donor and primary acceptor (bacteriopheophytin, H), and electron transfer from the primary to the secondary electron acceptor (ubiquinone). In particular, engagement of the accessory bacteriochlorophyll in primary charge separation is shown as an intermediate electron acceptor, and the initial free energy gap of approximately 40 meV, between the states P(+)B(A)(-) and P(+)H(A)(-) is estimated. The size of this gap is shown to be constant for the whole 230 ps lifetime of the P(+)H(A)(-) state. The ultrafast spectral dynamics features recorded in the visible range are presented against a background of results from similar studies performed for the last two decades.

Internal electrostatic control of the primary charge separation and recombination in reaction centers from Rhodobacter sphaeroides revealed by femtosecond transient absorption.

We report the observation of two conformational states of closed RCs from Rhodobacter sphaeroides characterized by different P(+)H(A)(-) --> PH(A) charge recombination lifetimes, one of which is of subnanosecond value (700 +/- 200 ps). These states are also characterized by different primary charge separation lifetimes. It is proposed that the distinct conformations are related to two protonation states either of reduced secondary electron acceptor, Q(A)(-), or of a titratable amino acid residue localized near Q(A). The reaction centers in the protonated state are characterized by faster charge separation and slower charge recombination when compared to those in the unprotonated state. Both effects are explained in terms of the model assuming modulation of the free energy level of the state P(+)H(A)(-) by the charges on or near Q(A) and decay of the P(+)H(A)(-) state via the thermally activated P(+)B(A)(-) state.

Linear and nonlinear magneto-optics of ferritin.

Measurements of Rayleigh light scattering and Cotton-Mouton (CM) effect are carried out at room temperature for 100 mM NaCl solutions of apoferritin/ferritin loaded with 0, 90, 100, 500, 700, and 1500 Fe atoms/molecule. Because of the spherical shape, ferritin macromolecule should not manifest magnetic anisotropy; however, in solution it shows the induced magnetic birefringence (CM effect) and changes in intensity of the scattered light components. The newly obtained data support the previously reported conclusions indicating that the deformation of linear optical polarizability induced in the ferritin by a magnetic field and the orientation of the induced magnetic dipole moment by this field are the main sources of the magneto-optical phenomena observed. Nevertheless, it is also found that the orientation of the permanent magnetic dipole moment contributes to both effects. The magnetic field induced changes in the light scattering and the CM effect theoretically depend on the linear magneto-optical polarizability, chi, on the nonlinear magneto-optical polarizability, eta, and square of the permanent magnetic dipole moment value of the macromolecule, mu(2). On the basis of the theory describing both effects as well as the experimental data, the values of the anisotropy of linear magneto-optical polarizabilities components, the values of the linear optical polarizability and its anisotropy, nonlinear magneto-optical polarizability and its anisotropy, are estimated. Also the magnetic dipole moment of the ferritin macromolecule is found. Interestingly, not all iron atoms in the ferritin are indicated to be in the superparamagnetic state, some of them occur in the diamagnetic form.

Mixed model studies on inheritance of reproductive traits in laying hens - a Bayesian approach.

Segregation analyses performed for many livestock species indicate a mixed inheritance model of reproductive traits. Additionally, depending on the population, a given trait can be determined by a number of genes with large effects. Genetic backgrounds of hatch-ability and fertility in poultry are still not known sufficiently. The objectives of this study are to verify the hypothesis on segregation of single genes (1 vs. 2) affecting fertility and hatchability and to estimate a heritability of these traits. Records from 2,040 and 2,015 dams from full-pedigreed strains of Rhode Island Red (R33) and New Hampshire (N88) from a pedigree farm were analyzed. The percentage of fertilized eggs and the percentage of the eggs hatched of fertilized eggs were registered for dams only. Fertility was checked by candling on the eighth day of incubation. To obtain a binomial phenotypic scale, 10 eggs per dam were included into the analysis. Animal single-trait threshold models were used for the analysis of data. The first model included the effects of 2 single genes, 2 fixed effects of year and season, additive polygenic effects, and permanent environmental effects. In the second model, only 1 single gene effect was included. Additionally, the analysis based on the polygenic threshold model was also performed. The Gibbs sampling procedure was used. The significance of single gene effects was verified by highest posterior density regions. The obtained results clearly gave evidence for the segregation of 1 major gene for hatchability in strain R33. Furthermore, the mixed inheritance model can also be suggested for fertility in this strain. After the analysis, the polygenic heritabilities were very low (<0.11), whereas major polygenic heritability ranged from 0.05 to 0.12.

Magneto-optics of Ferritin.

Measurements of Rayleigh light scattering, nonlinear light scattering in DC magnetic fields, and the Cotton-Mouton effect were carried out for 15 mM NaCl and water solutions of ferritin at room temperature. The spherical geometry of the molecule implies that it is optically isotropic. Such a macromolecule should not manifest magnetic anisotropy; however, in solution it shows induced magnetic birefringence (Cotton-Mouton effect) and changes in the intensity of the scattered light components. The analysis of the obtained results indicates the deformation of linear optical polarizability induced in the ferritin by a magnetic field as the main source of the magneto-optical phenomena observed. Light scattering and the CM effects theoretically depend on the linear magneto-optical polarizability, chi, and the nonlinear magneto-optical polarizability, eta. Using the theory describing the phenomena as well as the experimental data, the values of the anisotropy of linear magneto-optical polarizability components, chi(parallel) - chi(perpendicular) = -(1.3 +/- 0.7) x 10(-22) [cm3] (in SI units chi(parallel) - chi(perpendicular) = -(2.0 +/- 1.2) x 10(-33) [m3]), the linear optical polarizability, alpha = (alpha(parallel) + 2alpha(perpendicular))/3 = (3.9 +/- 1.0) x 10(-20) [cm3] (in SI units alpha = (3.52 +/- 0.09)x10(-4) [Cm2 V(-1)]), and its anisotropy, kappa(alpha) = (alpha(parallel) - alpha(perpendicular))/3alpha = -(0.06+/-0.03), nonlinear magneto-optical polarizability, eta = (eta(parallel) + 2eta(perpendicular))/3 = -(4.7 +/- 0.9) x 10(-30) [cm3 Oe(-2)] (in SI units eta = -(6.7 +/- 1.3) x 10(-18) [Cm4 V(-1) A(-2)]) and its anisotropy, kappa(eta) = (eta[parallel) - eta(perpendicular))/3eta = -(0.15 +/- 0.10), were deduced. Here alpha(parallel), eta(parallel), alpha(perpendicular), eta(perpendicular) are the optical and magneto-optical polarizability components along the parallel and the perpendicular axes of the axially symmetric molecule, respectively.

Comparison of power Doppler and velocimetry in predicting outcome of high-risk pregnancy.

OBJECTIVE: Prospectively evaluate semi-quantitative computer analysis of power Doppler (PD) signals in the placenta, tetal brain, lung, liver, kidney and spleen in high-risk pregnancies in relationship to perinatal outcome and also to compare tissue blood flow in the fetal brain and placenta with Doppler velocimetry. METHODS: PD signals were recorded in 180 high-risk pregnancies between 27 and 41 weeks of gestation. Images from PD angiographic scans were transmitted for computer analysis of pixel intensity. Mean flow signal intensity was recorded for each organ. The PD brain/lung ratio was calculated. The PD results were plotted on reference values and related to perinatal outcome. Middle cerebral (MCA), umbilical (UA) and uterine artery (Ut. A) velocimetry was also performed. RESULTS: High-risk pregnancies displayed lower PD signal intensity from the placenta, fetal lung, liver and kidney as compared to normals. However, the brain and spleen signals showed higher intensities suggesting increased tissue perfusion. PD signals from the fetal brain, lung, placenta and PD brain/lung ratio were correlated with perinatal outcome. The PD signal intensity from the fetal liver, kidney and spleen showed poor correlation with perinatal outcome. Fetal brain tissue blood flow showed better correlation with the outcome than MCA velocimetry. Placental tissue blood flow results were similar in predicting outcome to those obtained by means of UA and Ut. A velocimetry. CONCLUSION: In comparison with conventional Doppler velocimetry, computer analysis of PD signals, give similar results in the prediction of adverse perinatal outcome.

Modulation of primary radical pair kinetics and energetics in photosystem II by the redox state of the quinone electron acceptor Q(A).

Time-resolved photovoltage measurements on destacked photosystem II membranes from spinach with the primary quinone electron acceptor Q(A) either singly or doubly reduced have been performed to monitor the time evolution of the primary radical pair P680(+)Pheo(-). The maximum transient concentration of the primary radical pair is about five times larger and its decay is about seven times slower with doubly reduced compared with singly reduced Q(A). The possible biological significance of these differences is discussed. On the basis of a simple reversible reaction scheme, the measured apparent rate constants and relative amplitudes allow determination of sets of molecular rate constants and energetic parameters for primary reactions in the reaction centers with doubly reduced Q(A) as well as with oxidized or singly reduced Q(A). The standard free energy difference DeltaG degrees between the charge-separated state P680(+)Pheo(-) and the equilibrated excited state (Chl(N)P680)* was found to be similar when Q(A) was oxidized or doubly reduced before the flash (approximately -50 meV). In contrast, single reduction of Q(A) led to a large change in DeltaG degrees (approximately +40 meV), demonstrating the importance of electrostatic interaction between the charge on Q(A) and the primary radical pair, and providing direct evidence that the doubly reduced Q(A) is an electrically neutral species, i.e., is doubly protonated. A comparison of the molecular rate constants shows that the rate of charge recombination is much more sensitive to the change in DeltaG degrees than the rate of primary charge separation.

Analysis of single gene multitrait effects in livestock by the use of Gibbs sampling.

The paper presents a method of multivariate data analysis described by a model which involves fixed effects, additive polygenic individual effects and the effects of a major gene. To find the estimates of model parameters, the maximization of likelihood function method is applied. The maximum of likelihood function is computed by the use of the Gibbs sampling approach. In this approach, following the conditional posterior distributions, values of all unknown parameters are generated. On the basis of the obtained samples the marginal posterior densities as well as the estimates of fixed effects, gene frequency, genotypic values, major gene, polygenic and error (co)variances are calculated. A numerical example, supplemented to theoretical considerations, deals with data simulated according to the considered model.

Structural polymorphism of telomeres studied by photon correlation spectroscopy.

Photon Correlation Spectroscopy (PCS) was used to study the dynamics and structure of Tetrahymena telomeric sequence d(5'-TGGGGT-3')4. Two different modes were observed, corresponding to the following structures: intermolecular (tetramolecular) G-quadruplex and intramolecular (monomeric) G-quartet. Experimental values of translational diffusion coefficients DT were obtained for each structural form. The value of DT for the monomer equals to 1.4 x 10(6) (cm2/s), while for the tetramolecular structure, to 0.8 x 10(6) (cm2/s). The relative weight concentrations of these two forms were analyzed versus the concentration of NaCl varied from 10 mM to 500 mM. The values of experimentally determined diffusion coefficients were compared with those calculated assuming the "bead model" and with the atomic coordinates from the NMR and X-ray crystallographic data. For both structures the experimental and calculated values of DT were in reasonable agreement. In the entire NaCl concentration range studied, the contribution of the relative weight concentration of the monomeric telomere form changed from 85% for 10 mM NaCl to 60% for 500 mM NaCl.

Light gradients in spherical photosynthetic vesicles.

Light-gradient photovoltage measurements were performed on EDTA-treated thylakoids and on osmotically swollen thylakoids (blebs), both of spherical symmetry but of different sizes. In the case of EDTA vesicles, a negative polarity (due to the normal light gradient) was observed in the blue range of the absorption spectrum, and a positive polarity, corresponding to an inverse light gradient, was observed at lambda = 530 and lambda = 682 nm. The sign of the photovoltage polarity measured in large blebs (swollen thylakoids) is the same as that obtained for whole chloroplasts, although differences in the amplitudes are observed. An approach based on the use of polar coordinates was adapted for a theoretical description of these membrane systems of spherical symmetry. The light intensity distribution and the photovoltage in such systems were calculated. Fits to the photovoltage amplitudes, measured as a function of light wavelength, made it possible to derive the values of the dielectric constant of the protein, epsilons = 3, and the refractive index of the photosynthetic membrane for light propagating perpendicular and parallel to the membrane surface, nt = 1.42 and nn = 1.60, respectively. The latter two values determine the birefringence of the biological membrane, Deltan = nn - nt = 0.18.

Why does the light-gradient photovoltage from photosynthetic organelles show a wavelength-dependent polarity?

The light-gradient photovoltage from photosynthetic organisms and organelles is thought to arise from the primary charge separation in the reaction centers. The current explanation of the effect is the stronger excitation of the membrane side of a vesicle facing the light source than the one on the opposite side. Together with the known orientation of reaction centers, this explanation predicts unequivocally the polarity of the photovoltage. However, a polarity opposite to the one expected has often been reported. A dependence of the polarity on the wavelength has been published but no explanation was given (Gräber, P., and H.-W. Trissl. 1981. FEBS Lett. 123:95-99). Here we report on a theoretical treatment of light propagation and interference in pigmented and nonpigmented multilayers. A model calculation is carried out for a pair of membranes, demonstrating the wavelength-dependent light distribution as well as the relative photovoltage and its polarity. When the membranes contain no chromophores or when the absorption coefficient is low, the predicted polarity to that expected from a simple macroscopic absorption behavior. The model is tested by comparing new photovoltage data obtained at 532 nm as well as in the blue and red absorption bands of chlorophyll in chloroplasts. It is concluded that outside the main absorption bands the amplitude and polarity of the photovoltage is determined by the ratio of the refractive indices of the membrane and the medium.

Campylobacter pylori, NSAIDS, and smoking: risk factors for peptic ulcer disease.

Campylobacter pylori, nonsteroidal anti-inflammatory drugs, and smoking are associated with ulcer disease. To define further the role of these factors in ulcer disease, one hundred seven subjects presenting for endoscopy were tested for specific IgG and IgA antibodies to C. pylori, and questioned about nonsteroidal use and smoking. Sixty were dyspeptic patients, 28 were disease controls, and 19 were healthy asymptomatic volunteers. Considering all subjects, 81% (87/107) were either taking nonsteroidals or had antibody to C. pylori, and 32% of these (28/87) had an ulcer. Nineteen percent (20/107) were neither taking nonsteroidals nor had antibody to C. pylori, and none of these had an ulcer, p less than 0.01. Smokers, 41% (11/27), were more likely to have an ulcer than nonsmokers, 20% (16/80), p less than 0.05, but only because of the increased prevalence of ulcers in smokers who also had C. pylori, 73% (11/15) versus 27% (12/45) of nonsmokers with C. pylori, p less than 0.01. The use of nonsteroidals and antibody to C. pylori identify people at risk for ulcers. Smoking increases this risk in subjects with C. pylori. Absence of a history of nonsteroidal use and antibody to C. pylori identify individuals with a low probability of ulcer disease.

Oral ciprofloxacin and a monoclonal antibody to lipopolysaccharide protect leukopenic rats from lethal infection with Pseudomonas aeruginosa.

To study the treatment of infection with Pseudomonas aeruginosa, a leukopenic rat model was developed that closely mimics the pathogenesis of Pseudomonas infection in man. This model achieved approximately 90% mortality within 10 d of infection. Pseudomonas organisms were inconsistently shed from the feces despite gastrointestinal colonization (9 fecal v. 23 cecal cultures positive for challenge strain in 28 rats). Treatment of rats with oral ciprofloxacin at 40 mg/kg afforded complete protection. A suboptimal dose of ciprofloxacin (20 mg/kg), achieving peak levels of 0.31 micrograms/mL in serum and 26.3 micrograms/mL in stool, resulted in survival of 8 (40%) of 20 rats. Intraperitoneal administration of a monoclonal antibody directed at the lipopolysaccharide of the challenge strain of Pseudomonas resulted in survival of 5 rats (26%). The combination of the two increased the survival to 75% (15 of 20, P less than 0.05 compared to either treatment alone). Thus, the combination of suboptimal doses of ciprofloxacin and a monoclonal antibody appears to protect leukopenic rats from lethal infection better than either treatment alone.

The inhibitory effect of sodium alginate on antibiotic activity against mucoid and non-mucoid strains of Pseudomonas aeruginosa.

The exopolysaccharide of mucoid strains of Pseudomonas aeruginosa (MPA) is chemically similar to alginate, a common polysaccharide of seaweeds. Both polymers have been reported to decrease the diameter of the inhibition zones produced in antibiotic disc diffusion assays. In order to determine whether this phenomenon was due to reduced diffusion of the antibiotic in an agar matrix, or to inhibition of antibiotic activity by sodium alginate, we studied the effect of sodium alginate on the susceptibility of P. aeruginosa strains to antibiotics in disc diffusion assays, as well as in broth and agar dilution assays. Sodium alginate decreased the activity of the aminoglycoside antibiotics, amikacin and gentamicin, against two MPA strains and their non-mucoid derivatives in each of the assays. In broth dilution assays, increase of the calcium ion concentration likewise reduced aminoglycoside activity against P. aeruginosa, but not against Escherichia coli and Staphylococcus aureus. Sodium alginate caused no inhibition of the activity of piperacillin and carbenicillin. The reduction of aminoglycoside activity may have implications for the common failure of these antibiotics in the treatment of pulmonary infections caused by MPA in cystic fibrosis patients, but must be considered only an in-vitro phenomenon at present.

Antibacterial properties of 2-acetylpyridine-1-oxide thiosemicarbazones.

We have investigated the in vitro antibacterial activity of 13 2-acetylpyridine-1-oxide thiosemicarbazones and 5 thiosemicarbazides against 80 clinically significant bacterial cultures, including 13 isolates with known antibiotic resistance. Of the thiosemicarbazones tested, 5 had minimal inhibitory concentrations (MICs) of 0.25 microgram/ml for Neisseria gonorrhoeae isolates; 1 of these had an MIC range of 0.25-0.5 microgram/ml for the Neisseria meningitidis cultures, and 2 had MICs of 2 and 2-4 micrograms/ml for Staphylococcus aureus and Streptococcus faecalis isolates, respectively. Two of the thiosemicarbazides had MICs of 0.25 microgram/ml for N. gonorrhoeae, whereas 2 others had MICs of 2-4 and 4-8 micrograms/ml for S. aureus and S. faecalis isolates, respectively. The test compounds were ineffective against the gram-negative enteric cultures and the Pseudomonas isolates.

2-Acetylpyridine thiosemicarbazones XI: 2-(alpha-Hydroxyacetyl)pyridine thiosemicarbazones as antimalarial and antibacterial agents.

A series of 2-(alpha-hydroxyacetyl)pyridine thiosemicarbazones was synthesized as potential antimalarial and antibacterial agents. Their synthesis was achieved by the condensation of N4-mono- or N4,N4-disubstituted thiosemicarbazides with 2-(alpha-hydroxyacetyl)pyridine. The latter was prepared by selective bromine oxidation of (2-pyridinyl)-1,2-ethanediol. The new compounds show potent inhibitory activity against penicillin-sensitive as well as penicillin-resistant Neisseria gonorrhoeae (MIC, 0.5-0.004 micrograms/mL), against Neisseria meningitidis (MIC, 0.5-0.032 micrograms/mL), and Staphylococcus aureus (MIC, 0.5-2 micrograms/mL). Good in vitro antimalarial effects against Plasmodium falciparum (Smith strain; ID50, 6.7-38 ng/mL) were observed in most of these new agents, but only 3 of 12 compounds exhibit moderate in vivo activity against Plasmodium berghei. These new agents appear to be less toxic to the host and more water soluble than the corresponding 2-acetylpyridine thiosemicarbazones.