Use of clinical parameters for differentiation of gram-positive and gram-negative mastitis in dairy cows vaccinated against lipopolysaccharide core antigens

OBJECTIVE: To determine whether clinical parameters could be used to differentiate clinical mastitis (CM) caused by gram-positive bacteria from CM caused by gram-negative bacteria in dairy cows vaccinated against lipopolysaccharide core antigens. DESIGN: Case series. ANIMALS: 143 episodes of CM in 86 dairy cows in a single herd. PROCEDURE: Cows were examined at onset of CM, and 24 clinical parameters including rectal temperature, heart rate, rumen contraction rate, degree of dehydration, various udder and milk characteristics, lactation number, stage of lactation, and season of year were recorded. Milk production and milk constituent concentrations before onset of CM were obtained from Dairy Herd Improvement Association records. Values for cows with gram-negative CM were compared with values for cows with gram-positive CM. Logistic regression was used to identify important predictors of gram-negative CM. RESULTS: 64 (45%) CM episodes were caused by gram-negative bacteria and 79 (55%) were caused by gram-positive bacteria. Rumen contraction rate was significantly lower and milk protein percentage before onset of CM was significantly higher in cows with gram-negative, rather than gram-positive, CM. Logistic regression indicated that CM was more likely to have been caused by gram-negative bacteria if it developed during the summer, milk was watery, or rumen contraction rate was low. Sensitivity and specificity of the final regression model were 0.58 and 0.80, respectively. Predictive value of a positive result was 0.74 when proportion of CM episodes caused by gram-negative bacteria was assumed to be 50%. CLINICAL IMPLICATIONS: Results suggest that clinical observations do not allow accurate prediction of CM pathogens and should not be the sole criteria for deciding whether cows with CM are treated with antibiotics.     

Analysis of anti-U1 RNA antibodies in patients with connective tissue disease. Association with HLA and clinical manifestations of disease

Histatin 1 is a histidine-rich phosphoprotein present in human parotid saliva that possesses candidacidal activity and functions in mineralization by adsorbing to hydroxyapatite. The objective of the present study was to develop a system for recombinant production of histatin 1 and to examine the role of phosphorylation in the functional activities of this molecule. Native histatin 1 (containing a phosphoserine at residue 2) was purified from parotid saliva, whereas a bacterial expression system was used to produce a recombinant form of histatin 1 (re-Hst1) that lacked phosphorylated serine. Histatin 1 cDNA was inserted into the vector pGEX-3X, which expresses foreign genes as soluble fusion proteins attached to the carboxyl-terminus of glutathione S-transferase (GST). The GST/re-Hst1 fusion protein was isolated from cell lysates by affinity chromatography on glutathione (GSH)-Sepharose and digested with cyanogen bromide to separate re-Hst1 from the GST fusion partner. The digest was subjected to reversed-phase high-performance liquid chromatography on a C18 column, and re-Hst1 was eluted as a well-defined peak. The yield of re-Hst1 was 4 mg/L of bacterial culture. Amino-terminal sequencing and amino acid analysis confirmed the final product as re-Hst1. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that native histatin 1 and re-Hst1 had the same apparent molecular weights, while cationic PAGE showed that re-Hst1 was more basic. Phosphate analysis indicated 1 mol phosphate/mol of native histatin 1, while re-Hst1 lacked any detectable phosphate. Re-Hst1 demonstrated candidacidal activity comparable to that of native histatin 1, but displayed substantially lower binding to hydroxyapatite. These results show that phosphorylation of histatin 1 at residue 2 contributes significantly to its ability to bind to hydroxyapatite.     

A glycosylated Bence Jones protein and its autologous amyloid light chain containing potentially amyloidogenic residues

Amyloidosis is characterized by deposition of protein fibrils in various tissues. The wide variety of sequences of both amyloidogenic and non-amyloidogenic immunoglobulin light chains makes them a unique tool for addressing the importance of primary structure in the formation of insoluble fibrils. In this study, we have determined the primary structure of the kappa I immunoglobulin light chain from both the urinary Bence Jones protein and the deposited amyloid fibrils of a patient (MH) with primary amyloidosis. The sequence identity of urinary-excreted and tissue-deposited light chains excluded biclonality and somatic mutations and confirmed that the light chain existed in both a soluble and an insoluble form. Several residues have been previously reported to be significantly associated with amyloidogenic kappa chains. Many of these were found in the MH sequence, including Asp31, Asn45, Phe49, Gln55, His70, Asn/Gly93 and ProN/Val96, thereby supporting their potential role in fibrillogenesis. In addition, Asn20 and Pro60 of protein MH replaced the normally conserved Thr20 and Ser60. Asn20 was glycosylated in both the Bence Jones and the amyloid fibril protein MH. Cumulative effects of amyloid-associated residues and glycosylation might have rendered the immunoglobulin light chain MH prone to fibril formation.     

''Oncocheck'': an international external quality assessment scheme for immunoassays of tumor markers

Coccidia were isolated from 122 Belgian broiler farms without clinical coccidiosis. Shuttle programs including robenidin or nicarbazine in the starter (7-14 days) followed by an ionophore or diclazuril in the grower ration were most commonly used. Out of 215 coccidiosis-positive groups, 146 Eimeria acervulina, 65 E. maxima, and 88 E. tenella isolates were tested without further laboratory propagation in 17 sensitivity profiles. For each profile, oocytes were pooled from 9 +/- 4 farms (mean +/- SD) that used the same anticoccidial program and that belonged to the same integrated broiler operation. Each suspension contained an equal number of isolates and oocyst numbers from each farm tested. Each profile included an unmedicated uninfected group, an unmedicated infected group, and 11 medicated infected groups, consisting each of three replicates of three Ross chicks. Medication started at 8 days of age, and each inoculated bird received 50,000 sporulated oocysts at 10 days. Results were related to the anticoccidial program that had been in use. Chemical drugs showed the highest activity against Eimeria, whereas ionophores were less efficacious. Of the latter, monensin (110 ppm) was least active; narasin (70 ppm), salinomycin (60 ppm), and maduramicin (5 ppm) took an intermediate position, and lasalocid (90 ppm) was most active. A 50% improvement in weight gain was obtained in 7 to 10 out of 17 profiles with 100 + 8.35 ppm clopidol/methylbenzoquate (10), 125 ppm nicarbazin (9), 3 ppm halofuginone (8), and 1 ppm diclazuril (7). A 50% improvement in feed conversion was obtained in 7 to 11 profiles with nicarbazin (11), halofuginone (10), diclazuril (9), 33 ppm robenidine (9), clopidol/methylbenzoquate (7), and lasalocid (7). Based on relative oocyst output, the highest activity against E. acervulina was obtained with clopidol/methylbenzoquate (8/16); the highest activity against E. maxima was obtained with lasalocid (6/6), diclazuril (5/6), and halofuginone (5/6); and the highest activity against E. tenella was obtained with diclazuril (8/8), amprolium/ethopabate (5/8), halofuginone (4/8), maduramicin (4/8), and nicarbazin (4/8).     

Role of an inhibitory pyrimidine element and polypyrimidine tract binding protein in repression of a regulated alpha-tropomyosin exon

The lipogenic enzyme fatty acid synthase (FAS) is elevated in various human primary cancers and certain human cancer cell lines. FAS overexpression in human neoplasia has clinical relevance because of its association with tumor aggression and potential chemotherapeutic intervention. Here, we surveyed FAS in cell lines established from normal murine mammary epithelium (NMuMG) and from mammary tumors induced by either rodent polyoma (Py) virus or murine mammary tumor virus (MMTV). Western blotting revealed greater content of FAS in Py-transformed A1-1 and T1 than NMuMG or MMTV-transformed Mm5MT, RIIIMT and MMT060562. These data suggest that signaling events mediated by Py transformation may increase cellular amounts of FAS. Although FAS content was elevated to similar levels in A1-1 and T1, specific activities were significantly different as enzyme activity in T1 was 3-fold higher than A1-1. Likewise, FAS activity in NMuMG was about 0.5-fold higher than the MMTV-transformed lines, even though enzyme content was similar. Immunoprecipitation studies employing anti-phosphoamino acid antibodies followed by immunoblot analysis with anti-FAS antisera (and vice versa) were used to characterize the constitutive phosphorylation state of the enzyme. Phosphoserine and phosphothreonine residues were detected in the more active FAS from T1 and NMuMG, but not in the less active FAS from Mm5MT or A1-1. Discovery of phosphorylated FAS suggests that the enzyme may have more immediate control over lipogenesis than previously thought. High-dose (10-4 M) dexamethasone induced FAS content and activity in NMuMG and MMTV-transformed lines but not Py-transformed cells. Lower concentrations (10-8, 10-6 M) of dexamethasone also activated FAS but without concomitant elevation of its protein content, which was consistent with a phosphorylated form of FAS. Finally, cell lines were treated with the FAS inhibitor cerulenin: almost all breast cancer lines were growth inhibited at significantly lower amounts of drug than normal cell lineages, suggesting that FAS plays a greater role in viability of tumor cells than normal cells. Pretreatment with palmitate (a primary end-product of FAS) prior to cerulenin rescued A1-1 cells only slightly from growth inhibition, whereas pretreatment with oleate (a monounsaturated fatty acid synthesized from palmitate) synergized cerulenin's cytotoxic effects.     

Islet transplantation as a treatment for diabetes

BACKGROUND: The microvascular complications of diabetes are directly linked to hyperglycemia. Beta-cell failure is a critical factor in regulation of blood sugar levels. However, only a small proportion of persons with type 1 and type 2 diabetes obtain sufficient glycemic control to avoid complications. METHODS: There are two routes for beta-cell replacement, transplantation, and a mechanical beta cell equivalent. Beta-cell replacement therapy is a potential treatment modality, since diabetes is caused by beta-cell failure. RESULTS: An obvious path for glycemic control is some form of beta-cell replacement therapy. Successful islet transplantation is a difficult challenge, but current achievements with human pancreas transplants and islet allografts may greatly improve glycemic control. CONCLUSION: Beta-cell replacement therapy is an accepted treatment modality for diabetes.     

The composition of individual molecular species of plasma phosphatidylcholine in human pregnancy

The role of medical informatics in telemedicine is dependent on using the power of the computerized database to not only feed patient specific information to the health care providers, but to use the epidemiological and statistical information in the data base to improve decision making and ultimately care. The computer is also a powerful tool to facilitate standardizing and monitoring of care and when applied in continuous quality improvement methodology it can enhance the improvement process well beyond what can be done by hand. The coupling of medical informatics with telemedicine allows sophisticated medical informatics systems to be applied in low population density and remote areas.     

Future directions for resuscitation research. IV. Innovative advanced life support pharmacology

The topics discussed in this session include a partial review of laboratory and clinical studies examining the effects of adrenergic agonists on restoration of spontaneous circulation after cardiac arrest, the effects of varying doses of epinephrine, and the effects of novel vasopressors, buffer agents (NaHCO3, THAM, 'Carbicarb') and anti-arrhythmics (lidocaine, bretylium, amiodarone) in refractory ventricular fibrillation. Novel therapeutic approaches include titrating electric countershocks against electrocardiographic power spectra and of preceding the first countershocks with single or multiple drug treatments. These approaches need to be investigated further in controlled animal and patient studies. Epidemiologic data from randomized clinical outcome studies can give clues, but cannot document pharmacologic mechanisms in the dynamically changing events during attempts to achieve restoration of spontaneous circulation from prolonged cardiac arrest. Also, rapid drug administration by the intraosseous route was compared with intratracheal and intravenous (i.v.) drug administration. Many studies on the above treatments have yielded conflicting results because of differences between healthy hearts of animals and sick hearts of patients, differences in arrest (no-flow) times and cardiopulmonary resuscitation (CPR) (low-flow) times, different pharmacokinetics, different dose/response requirements, and different timing of drug administration during low-flow CPR versus during spontaneous circulation. The need to stabilize normotension and prevent rearrest by titrated novel drug administration, once spontaneous circulation has been restored, requires research. Most of the above topics require some re-evaluation in clinically realistic animal models and in cardiac arrest patients, especially by titration of old and new drug treatments against variables that can be monitored continuously during resuscitation.     

Hippocampal lesions cause forgetting in a spatial response task

Monkeys with lesions to the hippocampus and overlying cortex were impaired in making a spatially selective response on the basis of a spatial cue. Their impairment was even more severe on a task in which they were required to make spatial responses on the basis of cues which are not spatially distinct. A second experiment showed that once lesioned monkeys had been trained on a task with spatially distinct stimuli, they were initially able to perform accurately if the aspatial distinctiveness of the cue was reduced. However, their performance declined to chance over four to six trials. These results suggest that lesions to the hippocampus and overlying cortex may cause impairments in memory for the arrangement of visual scenes, including the spatial location of responses. Keywords: spiny neurons, direct pathway, indirect pathways, rat neostriatum