Polymer-polymer phase behavior.

Different polymers can be combined into a single material in many ways, which can lead to a wide range of phase behaviors that directly influence the associated physical properties and ultimate applications. Four factors control polymer-polymer phase behavior: choice of monomers, molecular architecture, composition, and molecular size. Current theories and experiments that deal with the equilibrium thermodynamics and non-equilibrium dynamics of polymer mixtures are described in terms of these experimentally accessible parameters. Two representative molecular architectures, binary linear homopolymer mixtures and diblock copolymers, exhibiting macrophase separation and microphase segregation, respectively, are examined in some detail. Although these model systems are fairly well understood, a myriad of mixing scenarios, with both existing and unrealized materials applications, remain unexplored at a fundamental level.

Polymersomes: tough vesicles made from diblock copolymers.

Vesicles were made from amphiphilic diblock copolymers and characterized by micromanipulation. The average molecular weight of the specific polymer studied, polyethyleneoxide-polyethylethylene (EO40-EE37), is several times greater than that of typical phospholipids in natural membranes. Both the membrane bending and area expansion moduli of electroformed polymersomes (polymer-based liposomes) fell within the range of lipid membrane measurements, but the giant polymersomes proved to be almost an order of magnitude tougher and sustained far greater areal strain before rupture. The polymersome membrane was also at least 10 times less permeable to water than common phospholipid bilayers. The results suggest a new class of synthetic thin-shelled capsules based on block copolymer chemistry.

Investigation of charge ratio variation in mRNA - DEAE-dextran polyplex delivery systems.

mRNA pharmaceuticals represent a new class of therapeutics, with applications, in cancer vaccination, tumour therapy and protein substitution. Formulations are required to deliver messenger RNA (mRNA) to the target sites where induction of genetic transfection following receptor mediated cell uptake & translation is required. In the current study, the cationic polysaccharide diethylaminoethylen (DEAE) - Dextran was selected as a model system carrier for the investigation of polyplex nanoparticle formation together with mRNA as a function of the molar ratio of the components. The structure of the mRNA/Dextran colloids was investigated as a function of the polymer-to-mRNA ratio and correlated with the biological activity determined by cellular transfection with luciferase coding mRNA. Dynamic light scattering (DLS), small angle x-ray scattering (SAXS), and small angle neutron scattering (SANS) with deuterium contrast variation were used to achieve structural insight into the systems. Similarly to previously investigated lipid based systems, colloidally stable particles with confined size were obtained with either excess of positive or negative charge. Highest activity was obtained with positive charge excess. From the scattering experiments information on the internal organization inside the polymer/mRNA systems was derived. Indication for the presence of structural elements in the length scale of ten to 20 nm were found in the excess of dextran, which could be due to either excess or particulate polymer. Information on the molecular organization of the mRNA nanoparticle products may provide a valuable basis for defining critical quality attributes of drug products for pharmaceutical application.

Cloning of a cGMP-gated cation channel from mouse kidney inner medullary collecting duct.

The cDNA sequence coding for the cGMP-gated cation channel expressed in the mouse kidney inner medullary collecting duct has been determined. The kidney cGMP-gated cation channel cDNA has an open reading frame of 2055 nucleotides and encodes a 685 amino acid protein. One cDNA clone is alternatively spliced thereby producing a deletion of 107 bp. Two differentially spliced 5' untranslated regions were determined by 5' RACE.

Adenosine 5'-triphosphate (ATP), the neurotransmitter in the prostatic portion of the longitudinal muscle layer of the rat vas deferens.

Suramin (1-100 microM) and alpha, beta-methylene adenosine 5'-triphosphate (AMPCPP, 39 microM), antagonized the motor activity induced by exogenous adenosine 5'-triphosphate (ATP) but not exogenous noradrenaline (NA) in the longitudinal musculature of prostatic (P) and epididymal (E) segments of the rat vas deferens. Likewise, application of these drugs reduced the fast component of the nerve-stimulated contraction in response to a single transmural electrical pulse in E and P. Suramin also blocked in a concentration-dependent fashion, the contractile responses to trains of 1.5, 5, 15 or 30 Hz transmural electrical pulses in P, while it did not affect those in E. AMPCPP obliterated responses to trains of 1.5, 5, and 15 Hz in P, while reducing these responses in E to a significantly lesser extent. Present results strongly support that ATP is the motor transmitter in P, while in E, ATP and NA are likely the co-transmitters responsible for the motor tone.

Liposomal daunorubicin in advanced Kaposi's sarcoma: a phase II study.

We report a non-randomized Phase II clinical trial to assess the efficacy and safety of liposomal daunorubicin (DaunoXome) in the treatment of AIDS related Kaposi's sarcoma. Eleven homosexual men with advanced Kaposi's sarcoma were entered in the trial. Changes in size, colour and associated oedema of selected 'target' lesions were measured. Clinical, biochemical and haematological toxicities were assessed. Ten subjects were evaluated. A partial response was achieved in four, of whom two subsequently relapsed. Stabilization of Kaposi's sarcoma occurred in the remaining six, maintained until the end of the trial period in four. The drug was generally well tolerated, with few mild symptoms of toxicity. The main problem encountered was haematological toxicity, with three subjects experiencing severe neutropenia (neutrophil count < 0.5 x 10(9)/l). There was no evidence of cardiotoxicity. In this small patient sample, liposomal daunorubicin was an effective and well tolerated agent in the treatment of Kaposi's sarcoma.

Diacetoxyscirpenol toxicity in pigs.

Purified diacetoxyscirpenol (DAS) mycotoxin fed to growing pigs at 0, 2, 4, 8, and 9 ppm for as long as nine weeks caused several lesions at all levels. The pigs developed multifocal, proliferative, gingival, buccal and lingual lesions. The small intestine had both glandular and mucosal cell hyperplasia. No other lesions were seen at necropsy or in any of the 19 tissues examined microscopically using haematoxylin and eosin stain. A one-way analysis of variance revealed statistically significant (P less than 0.05) decreased ration consumption and weight gain at all levels of DAS. Total ration refusal occurred at 10 ppm of DAS. There was no effect on the packed cell volume, haemoglobin concentration, total red blood cell count, total white blood cell count, 300 cell differential, terminal bone marrow smears or on the serum levels of aspartate transaminase, serum glutamic pyruvic transaminase and lactic dehydrogenase at any level of DAS. Based on the gross and microscopic lesions, decreased ration consumption and decreased weight gain the no-effect level was less than 2 ppm of DAS in the ration.

Response of fall-born calves to progesterone-estradiol benzoate implants and reimplants.

Two trials, involving 469 steer and heifer calves, were conducted to evaluate the practices of implanting or reimplanting suckling fall-born calves with progesterone-estradiol benzoate (PE) implants (100 mg of progesterone, 10 mg of estradiol benzoate). Calves having an average initial weight of 67 kg were randomly allocated within sex and location to remain either nonimplanted or to receive a PE implant in the late fall, in the spring just before green grass, or in both fall and spring. All animals grazed good- to excellent-quality native Bluestem range. Nonimplanted calves in Trial 1 gained .57 kg/d during the winter months and PE implants improved growth rate by 7% to .61 kg/d (P less than .05). Growth rate of calves on dry winter range in Trial 2 was only .20 kg/d, and was not affected by PE implants. Average daily gain of nonimplanted calves during the spring and summer months was .96 and .87 kg/d for trials 1 and 2, respectively. Performance of implanted calves during that period was improved an average of 4.3 to 10% (P less than .05) by PE implants in both trials regardless of when the calves were implanted. Growth rate of calves over the entire 8-mo trials was .66 kg/d for nonimplanted calves and was improved (P less than .01) to .70, .70 and .71 kg/d for calves on the two single-implant and reimplant schedules, respectively. Implanted calves gained an average of 10.4 kg more than nonimplanted calves during the study.

Evaluation of the agar gel immunodiffusion test for diagnosis of subclinical paratuberculosis in cattle.

Concurrent bacteriologic culture of feces and agar gel immunodiffusion (AGID) testing was performed on all cows and bred heifers over 14 months old in 10 dairy herds during a 32-month period to determine the effectiveness of the AGID test for the detection of subclinical paratuberculosis. Herds were sampled 5 times and, when possible, culled animals were tested again at slaughter. During 5 herd-wide samplings, Mycobacterium paratuberculosis was isolated from 139 fecal specimens obtained from 109 cattle. Results of the AGID test were simultaneously positive 40 of 139 times (28.8%). Thirty-six of the 109 cattle (33.0%) determined to be infected had a positive AGID test result at some point during the 5 herd-wide samplings. When results of tests performed at time of slaughter were included, 117 cattle were identified as infected by culture methods; 55 of these (47.0%) were AGID test-positive at some point during the study. The upper limit of the maximal false-positive rate for the AGID test was 2.1%. On the basis of colony counts from cultures, subclinically infected cows shedding higher numbers of M paratuberculosis in their feces were more likely to have positive AGID test results (P less than 0.0001). In known infected cattle, neither the culture nor AGID test results were consistently positive on repeated testing. Of 48 official calfhood paratuberculosis vaccinates tested as adults, 3 had positive AGID test results and in 1 of these, M paratuberculosis was also isolated from the feces, indicating that the rate of false-positive AGID test results in calfhood vaccinates is low.

Effect of zearalenone on dairy cows.

Doses of 99% pure zearalenone (0.0, 31.25, 62.5, 125.0, 250.0, or 500.0 mg) in gelatin capsules were given once a day per os to 18 nonpregnant, nonlactating, multiparous dairy cows for 2 consecutive estrous cycles. There was no effect (P less than 0.10) on serum progesterone concentrations, RBC, WBC, PCV, hemoglobin, and estrous cycle length. Differential cell counts, clinical health, and sexual behavior were not affected by the zearalenone. One cow from each of the groups given zearalenone and a control were euthanatized at the end of the study. The zearalenone had no effect on the terminal bone marrow smears and did not induce any gross lesions discernible at necropsy or any microscopic lesions in representative samples of 30 tissues/cow. Rectal palpation of the reproductive tracts once a week indicated that the corpora lutea were small in cows given zearalenone. There was a general trend to increased hemoglobin concentrations in cows given the larger doses of zearalenone. Zearalenone of and by itself does not seem to be an important factor in dairy cow health.

Effect of zearalenone on the fertility of virgin dairy heifers.

Eighteen cycling, virgin, Holstein heifers daily were given 250 mg of 99% purified zearalenone in a gelatin capsule orally, and 18 controls were given an empty gelatin capsule once a day. The study lasted through 1 non-breeding estrous cycle and the next 2 consecutive estrous cycles during which the 36 heifers were bred, using artificial insemination. Serum concentrations of progesterone and complete blood cell counts were determined throughout the study. The treated and control heifers had conception rates of 62% and 87%, respectively. There was no effect (P less than 0.05) on the serum concentration of progesterone or the complete blood cell counts. Three heifers, bred but not pregnant by the end of the study, were euthanatized and necropsied. The treated heifer did not have any zearalenone-attributable lesions, and there was no effect seen in the bone marrow smears. The remaining 33 heifers were sold as a herd, and the 31 pregnant heifers calved normally. There was no effect (P less than 0.05) on the sex ratio of the offspring, which were all clinically healthy. Zearalenone did lower the conception rate of the treated heifers (P less than 0.065).

Agar gel immunodiffusion test for diagnosis of clinical paratuberculosis in cattle.

During a 19-month period, the agar gel immunodiffusion (AGID) test was used as an aid in differential diagnosis of paratuberculosis in 51 cattle with a history of chronic weight loss and/or chronic diarrhea. Thirty-three cattle were AGID test-positive for paratuberculosis. Twenty-eight cattle (87.5%) yielded Mycobacterium paratuberculosis from bacterial culture of feces. Four cattle were confirmed as having paratuberculosis on necropsy. One had a negative fecal culture but was lost to follow-up. Thus, 32 of the 33 AGID test-positive cattle (96.9%) were confirmed as paratuberculous by culture or necropsy. Of the 18 cattle that were AGID test-negative, 17 (94%) also were culture-negative after 12 to 20 weeks' incubation of the culture tubes. Direct fecal smears from 29 confirmed paratuberculous cattle were examined. Twenty-three (79.3%) had smears positive for M paratuberculosis. Fifteen culture-negative cattle were examined by direct smear; the results were negative for all.

Comparison of collection techniques for quantitative urine culture in dogs.

The degree of bacterial contamination of 75 urine samples collected by voluntary voiding, catheterization, and cystocentesis was studied in 25 clinically normal dogs (14 females and 11 males) to evaluate the suitability of quantitative urine culture for diagnosis or urinary tract infection. Significant bacteriuria (greater than 100,000/ml) was not observed. Insignificant bacteriuria presumably caused by urethrogenital contaminants was detected in 44% of the urine samples collected by voluntary voiding, in 20% of the samples collected by catheterization, and in 12% of the samples collected by cystocentesis. The urine was sterile in 40% of the samples collected by voluntary voiding, in 80% of the samples collected by catheterization, and in 84% of the samples collected by cystocentesis. A bacteria count suggestive of bacteriuria (10,000--100,000/ml) was obtained in 1 sample collected by cystocentesis but was attributed to inadvertent penetration of a loop of intestine. Low bacteria counts were obtained in 4 (16%) samples collected by voluntary voiding, presumably as a result of urethrogenital and integumentary bacterial contaminants. It was concluded that urine samples properly collected from dogs by voluntary voiding, catheterization, or cystocentesis are suitable for detection of significant bacteriuria.

Flow study and SPECT imaging for the diagnosis of giant cell tumor of bone.

A case of giant cell tumor of bone is reported in which the blood flow study along with SPECT imaging improved the visualization and diagnosis of the abnormality.

Porcine abortion caused by Actinobacillus equuli.

Actinobacillus equuli was isolated from an aborted porcine fetus. Fetal lesions of peracute embolic septicemia were similar to those seen in septicemic actinobacillosis of older swine and other species. This case represented an individual rather than a herd abortion problem.

Effects of dietary zearalenone on finishing broiler chickens and young turkey poults.

A total of 168 broiler chickens (6 to 9 weeks of age) and 192 turkey poults (3-1/2 to 6-1/2 weeks of age) were fed purified zearalenone at levels of 0 (control), 10, 25 (turkeys only), 50, 100, 200, 400, and 800 mg/kg diet for a 3 week period to evaluate its toxicity. Dietary zearalenone had no effect on body weight gain, feed consumption, weights of liver, heart, spleen, and bursa of Fabricius, serum calcium, phosphorus, alkaline phosphatase, total protein and cholesterol, hematocrit, hemoglobin, red blood cell count, white blood cell count, and differential leukocyte count in both chickens and turkeys. Male broiler comb and testes weights were reduced by high levels of zearalenone. Female comb weight, broiler oviduct weight, and turkey testes and ovaries were unaffected by zearalenone. Histopathological examination of tissues revealed no change due to zearalenone. Male turkeys fed 400 and 800 mg/kg zearalenone had increased development of dewlaps and caruncles and exhibited considerable strutting behavior. Muscle and fat tissue from broilers fed 800 mg/kg zearalenone had no detectable estrogenic activity as measured by mouse uterine bioassay. These studies indicate that the effects of dietary zearalenone on finishing broilers and young growing turkey poults are minimal.

Effect of dietary zearalenone on reproduction of chickens.

Variable quantities of zearalenone (0, 10, 25, 50, 100, 200, 400, or 800 mg/kg diet) were incorporated into a practical laying hen diet and fed to 30-week-old White Leghorn females in egg production. During the 3 week pretest and 8 week experimental periods hens were inseminated weekly with .05 ml of pooled semen from males fed normal diets. Zearalenone was without effect on egg production, egg size, feed consumption, change in body weight, fertility, hatchability of fertile eggs, growth of progeny to 3 weeks of age, comb, weight, oviduct weight, heart weight, liver weight, spleen weight, egg shell thickness, Haugh units, blood hematology, serum calcium, serum inorganic phosphorus, and serum alkaline phosphatase. Zearalenone above 50 mg/kg of diet caused reduced serum cholesterol. In a reciprocal study, adult male New Hampshire chickens were fed diets containing 0, 100, or 800 mg/kg zearalenone for an 8 week period. Semen was collected and inseminated into White Leghorn females fed normal diets. Zearalenone was without effect on fertility or hatch of fertile eggs resulting from matings of these males. Zearalenone resulted in reduced serum inorganic phosphorus, serum cholesterol, and serum alkaline phosphatase in males. Histological examination of a number of tissues in both males and females revealed no changes due to zearalenone feeding. It is concluded that zearalenone up to 800 mg/kg of diet is without effect on reproductive performance of mature chickens.

Effects of T-2 toxin on reproductive performance and health of laying hens.

Purified T-2 toxin was fed to S.C.W.L. hens at levels of 0 (control), 0.5, 1.0, 2.0, 4.0 and 8.0 p.p.m. of an otherwise balanced diet. Feed consumption, egg production and shell thickness were significantly (P less than 0.05) decreased in hens fed 8 p.p.m. as compared with control hens. The fertility and progeny performance were not depressed by feeding T-2 toxin, but the hatchability of fertile eggs of hens fed 2 and 8 p.p.m. was significantly (P less than 0.05) lower than that of hens fed the control diet. The weights of liver, heart, gizzard and spleen were not influenced by T-2 toxin. Serum levels of alkaline phosphatase, LDH and uric acid of hens fed high concentrations of T-2 toxin were greater than those of control hens. SGPT in hens fed 8.0 p.p.m. was lower when compared with control hens. No outward changes in hematocrit, hemoglobin, erythrocyte, leukocyte and differential leukocyte counts were noted with feeding T-2 toxin. Most hens fed T-2 toxin developed oral lesions: circumscribed proliferative yellow caseous plaques at the margin of the beak, mucosa of the hard palate and angle of the mouth, and tongue. The incidence and severity of lesions were proportional to the dietary level of T-2 toxin. The only other lesion observed in necropsy examination at the end of the experiment was the small mucosal ulcer in the anterior portion of the gizzard in hens fed high levels of T-2 toxin. Microscostrointestinal tract, etc.) revealed no significant pathological change except the necrotic lesions in the gizzard and crop.

The failure of purified T-2 mycotoxin to produce hemorrhaging in dairy cattle.

A Holstein cow was intubated with 182 mg of 97% pure T-2 toxin (0.44 mg/kg of body weight) for 15 days. A dairy ration containing 50 mg/kg (50 ppm) of T-2 toxin was refused. A calf, born four days after onset of maternal treatment, was intubated with 26.2 mg of purified T-2 toxin (0.6 mg/kg of body weight) for seven consecutive days and then on alternate days for a total of 16 days. The calf was severely affected clinically by the T-2 toxin. The T-2 toxin failed to cause bovine hemorrhagic syndrome in either animal. Unspecific gastrointestinal lesions were noted in the cow but none were detected in the calf. In the calf, severe depression, hindquarter ataxia, knuckling of the rear feet, listlessness and anorexia were caused by the T-2 toxin.

Males, medical mutilation and the law: some recent developments.

Two recent decisions of the English Court of Appeal--Re J (Specific Issue Orders: Child's Religious Upbringing and Circumcision) [2000] 1 FLR 571 and Re A (Male Sterilisation) [2000] 1 FLR 549--raise serious issues relating to controversial matters involving law and medicine and deserve wider consideration than in their jurisdiction of origin. This article discusses the practical and policy implications of these cases and places them in the appropriate context.