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The aim of the present work was to assess the concentrations of metals in the abdominal muscle and exoskeleton of 3-year-old males and 4-year-old females and males of spiny-cheek crayfish (Orconectes limosus) collected from Lake Gopło. A total of 93 males and 35 females were collected in autumn (October 2014). The analyzes of heavy metals were conducted by means of atomic absorption spectroscopy with a PU9100X spectrometer. The content of mercury was determined using AMA 254 mercury analyser. As analyses indicated heavy metals accumulated in the muscle and exoskeleton in the following sequence: Zn > Cu > Pb > Mn > Ni > Hg and Mn > Pb > Zn > Ni > Cu > Hg, respectively. Statistically significant differences between 3- and 4-year-old males were found for all analyzed metals. Gender dependent differences were calculated only for Ni in the muscle tissue and for Mn and Hg in the exoskeleton. In comparison with the study carried out 2 years ago notably higher concentrations of Pb were found in the muscle and a higher content of Zn, Pb, Mn and Ni was determined in the exoskeleton.

Genetic selection that has been carried out for several dozen years has led to significant progress in poultry production by improving productive traits and increasing the profitability of broiler breeder and layer hen production. After hatching, broilers and layers differ mainly in feed intake, growth rate, efficiency of nutrient utilization, and development of muscles and adipose tissue. A key role can be played by hormonal mechanisms of appetite control in broilers and layers. The paper discusses the consequences of different growth rates resulting from long-term genetic selection on feed intake, efficiency of nutrient utilization, and development of muscles and adipose tissue, with particular consideration of the hormonal mechanisms of appetite control in broilers and layers. The information presented in this review paper shows that it would be worth comparing these issues in a meta-analysis., (© 2016 Poultry Science Association Inc.)

Chitin dihexanoate (DHCH) is the novel biocompatible and technologically friendly highly substituted chitin diester. Here we described optimization of DHCH and chitin dibutyrate (dibutyryl chitin, DBC) synthesis conditions (temperature and reaction time) to obtain desired polymers with high reaction yield, high substitution degree (close to 2) and appropriately high molecular weights. A two-step procedure, employing acidic anhydrides (hexanoic or butyric anhydride) as the acylation agent and methanesulfonic acid both as the catalyst and the reaction medium, was applied. Chemical structures of DBC and DHCH were confirmed by NMR ((1)H and (13)C) and IR investigations. Mechanical properties, thermogravimetric analysis, differential scanning calorimetry and biocompatibility (Neutral red uptake assay, Skin Sensitization and Irritation Tests) were assessed. Both polymers proved highly biocompatible (non-cytotoxic in vitro, non-irritating and non-allergic to skin) and soluble in several organic solvents (dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, acetone, ethanol and others). It is worth emphasizing that DHCH and DBC can be easily processed by solvent casting method and the salt-leaching method, what gives the opportunity to prepare highly porous structures, which can be further successfully applied as the material for wound dressings and scaffolds for tissue engineering., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The tissue factor (TF) - tissue factor pathway inhibitor (TFPI) axis plays a major role in hemostasis. Disorders of the coagulation system are commonly diagnosed with the help of screening tests such as prothrombin time (PT), activated partial thromboplastin time (aPTT), and plasma fibrinogen concentration (PFC). However, the effect of prebiotics on the hemostasis system has not been characterized in poultry yet. This study was designed to determine the effect of in ovo administration ofprebiotics on blood coagulation parameters of broiler chickens depending on their age. The study was conducted with 180 broiler chick embryos, the air cells of which were injected on day 12 of incubation with prebiotics (experimental groups: Bi2tos, DiNovoo and RFO) or physiological saline solution (control group). At 1, 21 and 42 days of rearing, blood was sampled from 15 broiler chickens from each group. An enzyme immunoassay was performed to determine plasma TF and TFPI levels, and PT, aPTT and PFC were determined in the chicken blood. We demonstrated that: 1) total TF levels increased with age in the experimental groups, 2) prebiotics had no significant effect on TF levels between the groups at a particular age, 3) total TFPI levels differed between both the type of in ovo injected substance and the broiler chicken age, 4) in the control group, PT and aPTT were found to increase with age whilst fibrinogen concentration decreased. The main conclusion from this pilot study is that total TF and TFPI levels change with age, however no clear patterns regarding TFPI were detected yet. The levels of PT, aPTT and PFC varied with the prebiotics administered in ovo as well as with the age of broiler chickens.

In this article, the synthesis of novel biopolymer, chitin dipentanoate (Di-O-Valeryl Chitin, DVCH) has been described. DVCH is a chitin derivative esterified with two valeryl groups at positions 3 and 6 of the N-acetylglucosamine units and it is soluble in common organic solvents like ethanol, methanol, acetone, dichloromethane, 1,2-dichloroethane, N,N-dimethylmethanamide, N,N-dimethylacetamide and ethyl acetate. Highly efficient synthesis (degree of esterification close to 2) of DVCH was achieved by employing a huge excess of valeric anhydride used as both the acylation agent and the reaction medium in the presence of perchloric acid as catalyst. Studies on the DVCH synthesis were aimed at finding optimal conditions (temperature, reaction time) to obtain DVCH with high reaction yield and desirable physicochemical properties. Biological data demonstrate that DVCH is non-cytotoxic in vitro and doesn't exert irritating or allergic effects to animal skin. Thanks to its filmogenic properties, it can be used to manufacture threads, foils, foams and non-woven materials. Moreover, DVCH can be easily processed by salt-leaching method to prepare highly porous structures exhibiting open-cell architecture, that can be further employed in wound dressing therapies and scaffolds for tissue engineering., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Intensive genetic selection of broiler breeders and layer hens for economically important production traits, which has been carried out for almost a century, resulted in considerable differences in the mechanisms of growth and development and, thus, in avian metabolism, both during embryogenesis and after hatching. Selection for meat production (broiler breeders) and eggs (layer hens) led to increased productivity but also brought about metabolic disorders. That intensive genetic selection of broiler breeders and layer hens is effective is seen, for example, in the differences in growth and development, metabolism of the yolk sac, hormones and lipids, gas exchange, and thermogenesis. Due to genetic proximity and different developmental mechanisms in broiler breeders and layer hens, avian embryos and chicks serve as excellent models for fundamental scientific research. This review paper discusses the consequences of different growth rates as a result of long-term genetic selection on embryonic development and metabolic rate of broilers and layers. The evidence presented herein indicates that it would be worth comparing these issues in a meta-analysis., (© 2015 Poultry Science Association Inc.)

In this study new biodegradable materials obtained by crosslinking poly(3-allyloxy-1,2-propylene succinate) (PSAGE) with oligo(isosorbide maleate) (OMIS) and small amount of methyl methacrylate were investigated. The porous scaffolds were obtained in the presence of a foaming system consisted of calcium carbonate/carboxylic acid mixture, creating in situ porous structure during crosslinking of liquid formulations. The maximum crosslinking temperature and setting time, the cured porous materials morphology as well as the effect of their porosity on mechanical properties and hydrolytic degradation process were evaluated. It was found that the kind of carboxylic acid used in the foaming system influenced compressive strength and compressive modulus of porous scaffolds. The MTS cytotoxicity assay was carried out for OMIS using hFOB1.19 cell line. OMIS resin was found to be non-toxic in wide range of concentrations. On the ground of scanning electron microscopy (SEM) observations and energy X-ray dispersive analysis (EDX) it was found that hydroxyapatite (HA) formation at the scaffolds surfaces within short period of soaking in phosphate buffer solution occurs. After 3h immersion a compact layer of HA was observed at the surface of the samples. The obtained results suggest potential applicability of resulted new porous crosslinked polymeric materials as temporary bone void fillers., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Drawbacks with the commonly used PMMA-based bone cements, such as an excessive elastic modulus and potentially toxic residual monomer content, motivate the development of alternative cements. In this work an attempt to prepare an injectable biomaterial based on isosorbide-alicyclic diol derived from renewable resources was presented. Two novel dimethacrylic monomers ISDGMA - 2,5-bis(2-hydroxy-3-methacryloyloxypropoxy)-1,4:3,6-dianhydro-sorbitol and ISETDMA - dimethacrylate of ethoxylated isosorbide were synthesized and used to prepare a series of low-viscosity compositions comprising bioactive nano-sized hydroxyapatite in the form of a two-paste system. Formulations exhibited a non-Newtonian shear-thinning behavior, setting times between 2.6 min and 5.3 min at 37°C and maximum curing temperatures of 65°C. Due to the hydrophilic nature of ISDGMA, cured compositions could absorb up to 13.6% water and as a result the Young's modulus decreased from 1,429 MPa down to 470 MPa. Both, poly(ISDGMA) and poly(ISETDMA) were subjected to a MTT study on mice fibroblasts (BALB/3T3) and gave relative cell viabilities above 70% of control. A selected model bone cement was additionally investigated using human osteosarcoma cells (SaOS-2) in an MTS test, which exhibited concentration-dependent cell viability. The preliminary results, presented in this work reveal the potential of two novel dimethacrylic monomers in the preparation of an injectable biomaterial for bone augmentation, which could overcome some of the drawbacks typical for conventional acrylic bone cement., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The aim of this study was to use biochemical markers to evaluate the quality of fresh and cryopreserved semen from the arctic fox (Vulpes lagopus). Twenty-three manually collected ejaculates were analysed for the main indicators of semen quality (sperm concentration and ejaculate volume). Sperm motility and percentage of morphologically normal and abnormal spermatozoa were determined according to the stage of cryopreservation (fresh--measurement A; equilibrated--measurement B; frozen/thawed--measurement C). Furthermore, the seminal plasma and supernatants were analysed after equilibration and freeze/thawing for the activity of the enzymes alkaline phosphatase (ALP), acid phosphatase (AcP), lactate dehydrogenase (LDH) and aspartate aminotransferase (AspAT), and for the activity of acrosin inhibitors (AP). The mean concentration of sperm was 625.1 million/cm3, and ejaculate volume averaged 1.6 cm3. Seminal plasma was characterized by the highest activity of alkaline phosphatase (3.43 x 10(3) U/l) and lowest activity of acrosin inhibitors (4.55 x 10(3) U/l). After equilibration, the supernatants showed the highest activity of acid phosphatase (94.9 U/l) and after freeze-thawing, they showed a high activity of lactate dehydrogenase (535.8 U/l) and aspartate aminotransferase (577.1 U/l), which indicates that these proteins had leaked from spermatozoa into the extracellular medium during the biotechnique of semen cryopreservation. In addition, several significant relationships were found between some indicators of semen quality and plasma and/or supernatant enzyme activity.

The aim of the research was to determine the effect of the season and the breeding region on the content of selected minerals (Zn, Cu, Mn, Fe, Pb, Cd) in the hair of dairy cows. The research material covered 114 Holstein-Friesian breed cows from three breeding centres in Poland, hereafterreferred to as A, B and C. Breeding centre A is located in Opolskie province, breeding centre B in Podlaskie province and breeding centre C in Kujawsko-Pomorskie province. The cows were kept in freestanding cowsheds with den boxes. Animal nutrition involved the use of the TMR system, considering division into nutrition groups. Hair was sampled in summer and in the period of winter and spring from the side of the body, directly behind the coastal arch. The Zn, Cu, Mn, Fe analysis was performed with atomic absorption spectrometry. The Pb and Cd content was assayed with the use of the electrothermal atomic absorption spectrometry. The results demonstrated the effect of season and breeding region on the level of the minerals and heavy metals assayed in the hair of cows being in the period of drying-off. The hair sampled in winter from cows from C demonstrated a higher concentration of most elements except for Fe than in material derived in summer. Hair sampled in winter from cows from the B centre showed higher Zn, Cu and Pb concentrations as compared with summer. Higher Cu, Mn, Fe and Pb concentrations were determined in hair sampled during winter from cows from A than in the material obtained in summer. The Cd content in the cow hair did not exceed the admissible norm, however, normal levels were exceeded for Pb in hair sampled in winter from cows from breeding centre C.

The objective of this study was to determine the effect of age and reproductive season on selected properties of semen from the arctic fox, Aloper lagopus L. The experiment used 40 ejaculates collected manually from 6 animals (3 foxes aged one year and 3 foxes older than three years). Statistically less semen (0.39 cm3) was collected from the young compared to the older animals, and the ejaculates obtained were characterized by higher concentration of spermatozoa (195.04 x 106/cm3). In turn, sperm acrosomal extracts from the older animals contained statistically more acrosin (6,4 mU/106 spermatozoa). In the sperm acrosomal extracts prepared during the first semen sampling, the mean acrosin activity did not exceed 2.3 mU/million spermatozoa. At subsequent semen sampling dates, the activity of the analysed enzyme increased to reach 7.72 mU/million spermatozoa. In the extracts obtained from the semen collected at the end of the breeding season of arctic foxes, the acrosin activity again reached a value obtained at the beginning of the season.

The aim of this work was to synthesize and investigate properties of a novel dimethacrylic monomer based on bioderived alicyclic diol--isosorbide. Its potential as a possible substitute of 2,2-bis[4-(2-hydroxy-3-methacryloyloxypropoxy)phenyl]propane (BISGMA), widely used in dental restorative materials and suspected for toxicity was assessed. The novel monomer was obtained in a three-step synthesis. First, isosorbide was etherified by a Williamson nucleophilic substitution and subsequently oxidized to isosorbide diglycidyl ether (ISDGE). A triphenyl phosphine catalyzed addition of methacrylic acid to ISDGE resulted in 2,5-bis(2-hydroxy-3-methacryloyloxypropoxy)- 1,4:3,6-dianhydro-sorbitol (ISDGMA). The monomer obtained was photopolymerized using camphorquinone/2-(dimethylamino)ethyl methacrylate initiating system. Next, compositions with triethylene glycol dimethacrylate (TEGDMA) were prepared and polymerized. Double bond conversion, polymerization shrinkage and water sorption of resulting polymers were determined. Selected mechanical (flexular strength and modulus, Brinell hardness) and thermomechanical (DMA analysis) properties were also investigated. BISGMA based materials were prepared as reference for comparison of particular properties.

The aim of this study was to determine fat and cholesterol content and the fatty acid profile in the meat of roach (Rutilus rutilus L.). Muscle samples for analyses were taken from the large side muscle of the fish body above the lateral line. The study involved 56 individuals caught in fall and spring. Analyses were carried out on 14 females and 14 males caught in each season. The percentage content of fat in the fish meat was determined with the modified method of FOLCH et al. (1957). The cholesterol content was determined with the modified Liebermann-Burchardt colorimetric method using a Shimadzu spectrophotometer (UV-VIS-NIR-3100). The fatty acid profile was determined with the gas chromatograph with a flame-ionization detector. Analyses indicated that the percentage content of fat was higher in the meat of roach caught in autumn (0.96% in females and 0.91% in males) than in the tissues of individuals from spring, which was 0.67% and 0.86%, respectively. The content of the total cholesterol was higher in autumn (64.17 mg 100g(-1)) than in spring (56.25 mg 100g(-1)), and these values significantly differed (p < 0.05). Statistical analyses indicated that there were no significant differences in the content of fat and cholesterol between females and males caught in both seasons. The total amounts of SFA, MUFA and PUFA ranged from 30.05% to 33.57%, from 39.53% to 47.64% and from 19.96% to 27.42%, respectively. Analyses of correlations between fatty acids from the n-3 and n-6 group indicated a positive and statistically significant relationship between C20:4 n-6 and C22:4 n-6 (p = 0.000, R = 0.9065) and C20:5 n-3 and C22:6 n-3 (p = 0.000, r = 0.8555). The ratio of n-3/n-6 was highest in the meat of roach caught in spring (0.50-0.81) than in autumn (0.50-0.77). The AI index ranged from 0.35 to 0.46 and the mean values of TI ranged from 0.41 to 0.54.

The aim of the study was to adapt a method to determine acrosin activity of human spermatozoa to arctic fox (Alopex lagopus L.) spermatozoa. We modified this method by reducing sperm count per sample from 1 divided by 10 x 10(6) to 25 divided by 200 x 10(3), incubation time from 180 minutes to 60 minutes, and Triton X-100 concentration in the reaction mixture from 0.01% to 0.005% per 100 cm3. It has also confirmed that arctic fox seminal plasma is rich in proteinases and their inhibitors. To completely abolish the inhibitory effect of seminal plasma on acrosin activity it is recommended to wash the spermatozoa four times. Benzamidine served an inhibitor of acrosin activity.

The aim of this study was to determine the chemical composition of roe deer meat considering the animals' sex and age and to estimate the content of amino acids in the meat from two selected groups of the animals, i.e. 2-3-year-old males and females. A further goal was to assess the biological value of proteins as compared to the FAO standard. The study has revealed that in proteins from the roe deer muscle tissue the content of exogenous amino acids (in g/100g) is higher by 20-30% on average comparing to the level of amino acids, in the FAO/WHO (1973) standard protein. Among the endogenous amino acids, the highest (in g/100 g of protein) and the lowest concentrations were found for glutamic acid and proline, respectively. The research has also shown that roe deer meat possesses a high content of protein and a relatively low content of fat.

Meat is an important source of animal protein but, at the same time, it includes saturated fatty acids, which makes it a potential cause of different cardiovascular diseases and still little is known about influence of age and sex on these parameters in roe deer muscles. The aim of this study was to determine the effect of age and sex on the CLA and other fatty acids content in the musculus longissimus lumborum in 67 roe deer (Capreolus capreolus L.). In the meat from the oldest individuals a higher content of CLA was noted (89.76 [mg/kg]) when compared to the fawns (42 [mg/kg]). In this research meat from roe deer does showed in general a higher percentage proportion of SFAs and MUFAs, but lower of PUFAs, than the meat from bucks. These results may provide an important source of information for consumers of roe deer meat because of differences between CLA and other fatty acids content depending on age and sex of the animals. Meat from roebucks is the most advantageous for dietary purposes.

The aim of this work was to compare the concentrations of calcium and phosphorus and Ca/P ratio in the meat of females and males of the roach (Rutilus rutilus L.) caught from the Brda River. The study involved 40 roach individuals caught in fall and spring (10 females and 10 males from each season). The muscle samples for analyses were taken from the large side muscle of the fish body above the lateral line. Ca concentrations were determined by atomic absorption spectrophotometry; P content was analyzed by the colorimetric method. Calcium concentration in the meat of analysed roach was higher in samples collected from fish caught in spring and equaled 1.82 g kg(-1) in females and 1.93 g kg(-1) in males. Values for individuals from autumn amounted to 0.83 and 1.10 g kg(-1), respectively. Statistically significant differences in calcium content were detected between individuals caught in different seasons, but samples taken from females and males caught within one season did not differ substantially. The mean value of P in the meat of analysed roach caught in spring was higher than in fish from autumn, and it was respectively 2.24 g kg(-1) in females and 2.30 g kg(-1) in males from spring, and 1.89 g kg(-1) in the tissue of females and 2.01 g kg(-1) in males in fish from autumn. The ratio of calcium to phosphorus in the meat of analysed wild roach ranged from 0.43:1 to 0.82:1. A negative and statistically significant correlation between Ca and P concentrations was found in the meat and the body length of analysed roach from the Brda River.

The Zn and Pb concentration was determined in 45 samples of bull semen from three Artificial Insemination Stations (A,B,C) differing in degree of environmental pollution. Bull semen was diluted with Biociphos-Plus to such an extent as to obtain 16 million sperm cells in an insemination dose. The research aimed at assessing the correlation between zinc, lead and other parameters influencing the semiological assessment of bull semen. The content of zinc and lead was determined with the use of the flame method and an absorption spectrophotometer (AAS). Positive, statistically significant correlations between zinc concentration and semen motility were observed in station A at r = 0.61 and station C at r = 0.614. In station B these correspondences were not statistically confirmed. Positive correlation at r = 0.583 was observed in case of zinc and the ejaculate volume only in station B. No statistically significant correlation was observed between lead concentration and the analyzed parameters.

To estimate the potential adverse consequences of rifampin therapy on cell-mediated immunity in tuberculosis, we measured in vitro lymphocyte responses to phytohemagglutinin, concanavalin A, pokeweed mitogen, and in vitro and in vivo responses to purified protein derivative tuberculin. Thirty-seven patients treated with therapeutic combinations containing rifampin were compared with 13 persons who had never received the drug. After initial improvement, responses to phytohemagglutinin became depressed in patients receiving rifampin for periods of 4 to 24 months. No significant changes were noted in lymphocyte responses to concanavalin A or pokeweed mitogen. In vitro and in vivo responses to purified protein derivative tuberculin were not altered. Because a favorable therapeutic outcome was achieved in all subjects, we concluded that rifampin does not have clinically significant immunosuppressive activity.

In an attempt to understand divergent observations regarding the in vitro lymphocyte responsiveness to antigens and mitogens, the frequency of cutaneous reactivity, and the relationship of these variables to clinical features of sarcoidosis, we examined cell-mediated immune responses in 75 untreated patients. In vitro lymphocyte responses to phytohemagglutinin and concanavalin A were significantly decreased among the group of patients with chronic active disease. In vitro lymphocyte responses to streptococcal antigen paralleled the patient's cutaneous reactivity; however, when compared to healthy persons, patients with sarcoidosis responded significantly less often to streptococcal skin tests. Twelve of 13 persons expected to be tuberculin positive by clinical history were found to be dermally reactive, and 11 these 12 exhibited positive in vitro lymphocyte responses to purified protein derivative. Among the remaining 62 patients, failure to respond to all stimuli in vivo and in vitro occurred in only 10. No evidence was found to support the general view that sarcoidosis is characterized by anergy to specific antigens. The patterns of responses to mitogens were neither characteristic nor unique for sarcoidosis. In patients in whom diminished specific cellular immune reactivity was observed, no correlation was evident with specific clinical features of the disease.

Immunoelectrophoretic analyses were performed on a cell extract and culture filtrate of Mycobacterium tuberculosis, H37Rv strain using homologous polyvalent goat antisera. General similarity between the cell extract and culture filtrate antigens was found. Significant variation between two anti-culture filtrate antisera was observed. These antigens and antisera also differed significantly from previously described reference reagents. These findings demonstrate the necessity of using reference materials for precise identification of mycobacterial antigens in comparative studies.

The in vitro lymphocyte transformation test was compared to the skin test at intervals after aerogenic administration of bacille Calmette-Guerin vaccine to monkeys, and also at monthly intervals after aerogenic challenge of monkeys vaccinated and not vaccinated with virulent strain H37Rv of Mycobacterium tuberculosis. Individual responses varied, but several consistent patterns of sensitivity development could be discerned. The lymphocyte transformation test was more sensitive, and often positive when the skin test was negative, doubtful, or feeble. Conversion to tuberculin reactivity was detected by lymphocyte transformation in vitro earlier in the disease or after vaccination, and persisted longer after sensitivity to the skin test had waned or after the animals had become anergic by the skin test. Monkeys not vaccinated, but challenged, developed larger in vitro skin reactions and responses than animals that were either vaccinated and challenged or only vaccinated; however, the unvaccinated, challenged monkeys developed anergy to tuberculin and progressive disease more rapidly than other groups, and their cells became less responsive to phytohemagglutinin in vitro.

Blastogenic responses in vitro to phytohemagglutinin and pokeweed mitogen were examined in microcultures of peripheral blood lymphocytes from a group of 12 healthy, long-term marihuana smokers and a group of matched control subjects. With either mitogen, no significant difference in cellular incorporation of (3H)thymidine was noted between the groups. These results were interpreted to indicate that the functional status of blood lymphocytes was not altered by long-term smoking of marihuana.

Peripheral blood lymphocytes from 60 black patients with Mycobacterium tuberculosis disease proved by culture and 100 healthy black persons were HLA phenotyped using a standard lymphocytotoxicity test. Statistical analysis of the results showed that only Bw15 antigen had a significantly increased frequency among patients with tuberculosis when they were compared to healthy persons. Furthermore, examination of disease manifestations among patients with tuberculosis demonstrated that patients with Bw15 had significantly more far-advanced pulmonary disease with cavitation than did patients without Bw15. These observations suggest that Bw15 may influence both susceptibility to tuberculosis and the course of the disease.

Comparisons were made of the yield, chemical content, and biological activity of filtrates and extracts obtained by sonic and pressure cell disruption of bacilli from 4- and 8-week-old Proskauer and Beck cultures of the H37Rv strain (TMC no. 102) of Mycobacterium tuberculosis. The culture filtrates were dialyzed, freeze-dried, reconstituted in saline, and sterilized by membrane filtration. The viable bacilli were washed and resuspended in distilled water and subsequently disrupted either by sonication in the cold for 15 or 30 min or by treatment at 20,000 or 40,000 lb/in2 in a pressure cell. The resulting extracts were clarified by centrifugation, concentrated, and sterilized by filtration. All preparations were adjusted to contain 10 mg of solids (dry weight)/ml and were analyzed quantitatively for protein, deoxyribonucleic acid, ribonucleic acid, polysaccharide, and lipid content. Separation patterns obtained by gradient acrylamide gel electrophoresis, as well as by one- and two-dimensional immunoelectrophoresis, provided the basis for qualitative comparisons of the culture filtrates and cell extracts. Three-point dose-response curves also were used to compare the preparations for skin test reactivity in BCG-vaccinated guinea pigs. It was concluded that, although there were no consistent differences in chemical content or biological activity between the preparations, a 15-min sonic treatment appeared to be the most suitable method for preparation of bacillary extracts based on yield of active components and ease of preparation.

Fractions of cell sonicates from the H37Rv strain of Mycobacterium tuberculosis prepared by gradient acrylamide gel electrophoresis (GAGE) were examined by immunoelectrophoretic analysis and tested for biologic reactivity and antigenic specificity. Each was tested for the ability to elicit delayed type hypersensitivity reactions in the skin of guinea pigs sensitized with M. tuberculosis, M. kansasii, M. intracellulare (Battey), M. scrofulaceum (Gause), and M. fortuitum. Peripheral lymphocytes from persons infected with mycobacteria were also exposed to fractions, and the increase in [3H]thymidine incorporation into DNA was used as an index of stimulation (lymphocyte transformation). Only 4 fractions (6, 11, 12, and 16) were found to be significantly more specific than the unfractionated sonicate, and none was monospecific for M. tuberculosis. The results of fused rocket immunoelectrophoresis suggested that incomplete separation of antigens and/or shared group-specific multideterminant antigens may have accounted for the observed cross-reactions.

The length of immunoelectrophoretic separation patterns obtained with a reference mycobacterial antigen was reduced significantly when a continuous buffer system was used in place of the recommended discontinuous system. Varying the supporting medium by using 1% concentrations of different agar preparations in the discontinuous system also affected the separation length and resolution of the reference pattern. Measurement of the electrical resistivity of the agar medium provided an explanation of these differences on the basis of current density or field intensity applied per slide. Variation in the reference pattern obtained in differently designed electrophoresis chambers also was attributed to differences in field intensity. To avoid these obvious alterations in the separation pattern of the reference antigen, it was suggested that separation procedures described originally for the reference system be used without modification.

Rose, Michael J., Jr. (Veterans Administration Hospital, Washington, D.C.), Stephen A. Aron, and Bernard W. Janicki. Influence of nonionic surfactants on bacteriophage infections. I. Effect of Triton WR-1339 on T(2) coliphage infection. J. Bacteriol. 87:933-938. 1964.-Plaques formed by T(2) phage on cultures of Escherichia coli strain B from nutrient broth which contained 0.5% Triton WR-1339 were considerably smaller than those formed on cultures from control broth. Pretreatment of phage particles with Triton did not result in small plaques. Results of population studies showed that treated and control cultures had the same growth rates. Similarly, characteristic physiological reactions of treated cultures were the same as those of the controls. Investigation of the phage growth cycle demonstrated that neither the adsorptive phase nor the latent period was affected by cultivation of the host in Triton. However, the burst size of treated cultures was significantly less than that of control cultures. Analysis of the one-step growth curve data showed that Triton did not inhibit bursting of infected cells. It was concluded that the smaller plaque size was a reflection of a reduced yield of mature virulent phage particles from the Triton-treated cultures.

Incubates of morphine with serum globulins obtained from sera of rabbits immunized with a morphine-bovine serum albumin conjugate produced immediate Schultz-Dale contractions when added to superfused, electrically stimulated guinea-pig ileal strips. Incubates of morphine with Krebs-Henseleit solution produced relaxation and depression of tone, and inhibition of electrically induced contractions. It is concluded that the spasm of guinea-pig ileum produced by incubates containing morphine-binding serum globulins and morphine resulted from transient passive sensitization and an acute anaphylactic type of response.

Guinea pigs given three subcutaneous injections of triton WR-1339, 300 mg./kg., at 3 day intervals develop an intense lipemia as evidenced by marked increases in serum phospholipide, free cholesterol, ester cholesterol, neutral fat, and total fatty acids. The total fatty acid content of the lung, liver, and spleen, however, remains unchanged. No qualitative change in the fatty acids of the lung and liver from treated animals could be detected by the methods used. The injection of triton was found to diminish slightly the cholesterol content of the adrenal glands. The implications of these findings are discussed in relation to an understanding of the mechanism involved in the therapeutic action of triton in experimental tuberculosis.