Combined center of rotation of angulation-based leveling osteotomy and tibial tuberosity transposition: An ex vivo mechanical study.

OBJECTIVE: To describe the technique of combined center of rotation of angulation (CORA)-based leveling osteotomy (CBLO) with tibial tuberosity transposition (TTT) and to compare the load to failure between CBLO combined with TTT and CBLO or TTT alone. STUDY DESIGN: Ex vivo study. SAMPLE POPULATION: Twelve pairs of cadaveric pelvic limbs. METHODS: Six pairs of cadaveric tibia were tested in each group (CBLO-TTT versus CBLO) and (CBLO-TTT versus TTT) with each limb randomly assigned to a treatment group. Construct stability was determined by applying a tensile force to each patellar tendon until failure occurred. Load at failure and mode of failure were recorded for each specimen. RESULTS: No difference in mean load to failure was identified between CBLO-TTT (897 N) and CBLO alone (943 N) (P = .81). There was also no difference in the mean load to failure between the CBLO-TTT (928 N) and TTT alone (1046 N) (P = .12). CONCLUSION: Performing a TTT in combination with a CBLO does not weaken the construct failure to load when compared with each procedure performed alone. CLINICAL SIGNIFICANCE: A combined CBLO and TTT could be considered a viable option for concurrent management of a cranial cruciate ligament deficient stifle and medial patella luxation.

Protection of sheep by vaccination against experimental challenge with Leptospira borgpetersenii serovar Hardjo and L. interrogans serovar Pomona.

AIMS: To evaluate a multivalent leptospiral and clostridial vaccine for prevention of renal colonisation and urinary shedding in sheep, following experimental challenge with New Zealand strains of Leptospira borgpetersenii serovar Hardjo type Hardjobovis and L. interrogans serovar Pomona. METHODS: Two separate but similarly designed studies were conducted. In both studies, Romney-cross lambs, aged 9-11 weeks, were randomly allocated to a vaccinated group and a control group. Vaccinated lambs each received two 1.5-mL S/C doses of a multivalent leptospiral and clostridial vaccine, 4 weeks apart, and animals in the control groups received the same dose of saline. Groups of 12 vaccinated and 12 control lambs were randomly selected in each study for challenge with serovars Hardjo or Pomona. Challenge was initiated 16 weeks following the second vaccination with three daily doses of live leptospires by intranasal and conjunctival routes. Following challenge, urine samples were collected weekly for 6 weeks, for dark field microscopy and leptospiral culture; 6 weeks after challenge the lambs were slaughtered and kidneys collected for leptospiral culture. RESULTS: In lambs challenged with serovar Hardjo, 8/12 unvaccinated lambs had ≥1 urine or kidney sample that was positive for leptospires following culture, compared with 0/12 lambs in the vaccinated group (p=0.001). In lambs challenged with serovar Pomona, 9/12 unvaccinated lambs had ≥1 urine or kidney sample that was positive following culture, compared with 0/12 lambs in the vaccinated group (p<0.001). Prevention of renal colonisation and urinary shedding, expressed as the prevented fraction, was 100 (95% CI=61.7-100)% and 100 (95% CI=68.3-100)% against challenge with serovars Hardjo and Pomona, respectively, at 4 months after vaccination. CONCLUSIONS AND CLINICAL RELEVANCE: Use of a multivalent leptospiral and clostridial vaccine demonstrated protection against challenge from New Zealand strains of serovars of Hardjo and Pomona 4 months after vaccination in lambs first vaccinated at 9-11 weeks of age. Further studies are required to assess the duration of immunity against challenge in sheep.

Factors associated with colostrum quality in individual cows from dairy herds in the Waikato region of New Zealand.

AIMS: To examine associations between various cow-level factors and quality of first-milking colostrum (measured as Brix), and to evaluate herd-level associations between vaccination against calf diarrhoea and colostrum quality, in cows from dairy herds in the Waikato region of New Zealand. METHODS: A single colostrum sample was collected, by complete udder evacuation, from each of 20 cows from 29 dairy herds in the Waikato region of New Zealand during the 2016 spring calving period. Vaccination pre-partum with a calf diarrhoea vaccine was used in 15 herds. Each colostrum sample was tested using a digital Brix refractometer. The body condition score of each cow was recorded at the time of sample collection and farmers provided records of clinical mastitis and facial eczema from the previous 12 months, as well as the age and breed of cows. Associations between cow-level variables in non-vaccinated herds and Brix were examined using a multivariable linear mixed model and estimated marginal means obtained for different categories. RESULTS: Mean Brix of 281 samples from cows in non-vaccinated herds was 18.7 (SD 0.26)%; 63/281 (22.4%) samples had Brix ≥22% and 152/281 (54.1%) had Brix ≥18%. Mean Brix of colostrum samples from cows aged ≥6 years (20.2 (95% CI=19.1-21.2)%) was higher than for samples from 2-year-old cows (18.6 (95% CI=17.3-19.9)%) (p=0.005). Colostrum that was collected at the first milking on the day of calving had higher Brix (20.0 (95% CI=19.1-20.9)%) than colostrum collected from cows that calved the previous day (17.5 (95% CI=16.5-18.4)%) (p<0.001). Mean Brix of colostrum samples from cows which produced ≥8 L (18.2 (95% CI=17.1-19.2)%) tended to be lower than from cows which produced <8 L first-milking colostrum (19.1 (95% CI=18.3-20.0)%) (p=0.08). Among vaccinating herds, 9/15 (60%) had ≥60% colostrum samples with Brix ≥18% compared with 4/14 (29%) of non-vaccinating herds (p=0.04). CONCLUSIONS AND CLINICAL RELEVANCE: Colostrum quality, as measured by Brix, was associated with the total volume of first-milking colostrum, interval from calving to colostrum collection and cow age. Vaccination against calf diarrhoea was associated with a higher proportion of colostrum samples with adequate Brix. Careful selection of colostrum donor cows should ensure newborn calves are fed adequate quality colostrum which should be beneficial in preventing failure of passive transfer of IgG. Testing of colostrum from individual cows with a Brix refractometer is advocated for the selection of colostrum for feeding newborn calves.

Prevention of fetal infection in heifers challenged with bovine viral diarrhoea virus type 1a by vaccination with a type 1c or type 1a vaccine.

AIMS: To evaluate a vaccine containing type 1c bovine viral diarrhoea (BVD) virus for prevention of fetal infection in pregnant heifers when challenged with New Zealand BVD virus type 1a 6 months after vaccination, compared to unvaccinated heifers and heifers vaccinated with a vaccine containing type 1a BVD virus. METHODS: Fifty five crossbred Friesian heifers, free from BVD virus and antibody, were randomly allocated to three groups. Twenty five heifers were vaccinated twice with a vaccine containing type 1c BVD virus (T1c group), and 10 heifers with a vaccine containing type 1a BVD virus (T1a group), and 20 heifers were unvaccinated (NC group). After oestrus synchronisation the heifers were bred by artificial insemination followed by natural bull mating. Six months after booster vaccination 15 heifers from the T1c group, eight from the T1a group, and 15 from the NC group, were exposed to four calves that were persistently infected with type 1a BVD virus, for 4 weeks. At the beginning of the challenge phase 36/38 heifers were 72-74 days pregnant and 2/38 heifers were approximately 53 days pregnant. Approximately 52 days after the start of the challenge the heifers were subjected to euthanasia and fetal tissues were collected for the detection of BVD virus by ELISA in fetal heart blood and PCR in fetal tissues. RESULTS: Based on PCR results, BVD virus was detected in 15/15 fetuses in the NC group, compared to 4/14 fetuses in the T1c group and 3/8 fetuses in the T1a group. The proportion of BVD virus-positive fetuses was lower in both vaccinated groups compared to the NC group (p<0.002), but there was no difference in proportions between the vaccinated groups (p=1.00). Fetal protection, expressed as the prevented fraction, was 71.4 (95% CI=41.9-91.6)% and 62.5 (95% CI=24.5-91.5)% for the T1c and T1a groups, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: The vaccines containing killed type 1c and type 1a BVD viruses significantly reduced fetal infection following challenge with a New Zealand type 1a BVD virus. Prevention of fetal infection by vaccination may not be 100%, and the risk of persistently infected calves being born to some vaccinated cattle should be acknowledged and managed as part of a BVD control programme.

Precipitants of brain infarction. Roles of preceding infection/inflammation and recent psychological stress.

BACKGROUND AND PURPOSE: Antecedent febrile infection and psychological stress are described as predisposing risk factors for brain infarction. We examined the temporal relationship between preceding infection/inflammation and stroke onset as well as the role of recent psychological stress as a potential precipitant for brain infarction. METHODS: In this case-control study, a standardized evaluation including a signs/symptoms-based questionnaire was used to characterize the prevalence and timing of recent prior (< 1 month) infectious and inflammatory syndromes in 37 adults with acute brain infarction, 47 community control subjects, and 34 hospitalized nonstroke neurological patient controls. Recent psychological stress was measured with scales of stressful life events and negative affect. RESULTS: The prevalence of infection/inflammation was significantly higher in the stroke group only within the preceding 1 week compared with either community control subjects (13/37 versus 6/47, P < .02) or hospitalized neurological patient controls (3/34, P < .02). Upper respiratory tract infections constituted the most common type of infection. A substantial proportion of stroke patients with preceding (< 1 week) infection/inflammation (5/13) had no accompanying fever or chills. There were no significant differences between the stroke and control groups in the levels of stressful life events within the prior 1 month or in negative-affect scale scores within the prior 1 week. CONCLUSIONS: Our data suggest that both febrile and nonfebrile infectious/inflammatory syndromes may be a common predisposing risk factor for brain infarction and that the period of increased risk is confined within a brief temporal window of less than 1 week. Results of this study argue against a role for recent psychological stress as a precipitant for cerebral infarction.

Extragenital Mycoplasma hominis infections in adults: emphasis on immunosuppression.

Mycoplasma hominis, a commensal organism that is potentially pathogenic both in maternal perinatal and in neonatal infections, also causes nongenitourinary infections in adults. We reviewed the clinical features of cases from the literature and emphasized recent cases. Infection sites were classified as blood, vascular sites, wounds, central nervous system, joints, and respiratory tract. Twenty-one of 31 newly summarized cases and 32 of 67 overall cases were associated with immunosuppression and/or hypogammaglobulinemia. Clinical suspicion, use of appropriate culture media, and determinations of gamma-globulin levels and of antibodies specific to Mycoplasma are indicated in the proper clinical setting. These types of M. hominis infection appear to be linked to suppressed cell-mediated immunity and/or hypogammaglobulinemia.

Septic arthritis and bacteremia due to Mycoplasma resistant to antimicrobial therapy in a patient with systemic lupus erythematosus.

We report a case of septic arthritis caused by Mycoplasma hominis in a patient with systemic lupus erythematosus. The infection started as monarthritis but spread to at least four joints despite apparently suitable therapy with various antimicrobial agents, including doxycycline, clindamycin, and ciprofloxacin; subsequent bacteremia was documented. Control was ultimately achieved with use of the experimental fluoroquinolone temafloxacin in combination with doxycycline administration, arthroscopic drainage of a persistently infected joint, several intravenous infusions of immunoglobulins (which led to increases in levels of antibodies specific to M. hominis), and discontinuation of corticosteroid therapy. Antimicrobial susceptibility testing of various mycoplasmal isolates showed the presence of the tetM gene, disparity between susceptibility to tetracycline and doxycycline, and increasing resistance to most antimicrobial agents used (including to fluoroquinolones before clinical use), although the patient ultimately had a favorable clinical response to treatment with combined modalities.

Epstein-Barr virus induces a unique pyrimidine deoxynucleoside kinase activity in superinfected and virus-producer B cell lines.

Epstein-Barr (EB) virus induces a new pyrimidine deoxynucleoside kinase [thymidine kinase (dTk)] activity in Raji B lymphocyte cells after superinfection. This dTk activity is also present in small amounts in the HR-1 virus-producer cell line and in larger amounts in the B95-8 virus-producer line. The dTk activity induced by EB virus coelutes from DEAE-cellulose columns with deoxycytidine kinase (dCk) activity and elutes as a broad peak well separated from the large peaks of cellular dTk and dCk activities. This EB virus-induced pyrimidine deoxynucleoside kinase activity from HR-1 cells differs from cellular kinases in most basic biochemical properties but shares certain properties with the herpes simplex virus dTk.

Hypomethylation of Epstein-Barr virus DNA in the nonproducer B-cell line EBR.

The conversion of the Epstein-Barr virus-negative Ramos cell line has previously been shown to result in an Epstein-Barr virus-positive non-virus-producer cell line, EBR. We report here that Epstein-Barr virus DNA from EBR alone among several cell lines examined was totally unmethylated at three of four sites containing guanine plus cytosine which were tested. This is in direct contrast to reports of high degrees of methylation in the DNAs of other animal viruses, including herpesviruses, isolated from cells in which the viral genome is expressed at a low level.

Incorporation into DNA of the base analog 2-aminopurine by the Epstein-Barr virus-induced DNA polymerase in vivo and in vitro.

The Epstein-Barr virus (EBV)-induced intracellular DNA polymerase was assayed in vitro for the ability to utilize the mutagenic nucleotide analog 2-aminopurine deoxyribose triphosphate (d2apTP), incorporating it as the corresponding monophosphate into DNA or poly[d)(A-T)] template. Bacteriophage T4, lymphocyte alpha, and the EBV particle-associated DNA polymerases were assayed simultaneously for direct comparison. Unlike these three polymerases, which were capable of distinguishing between d2apTP and dATP with a strong preference for the latter, the EBV-induced DNA polymerase only weakly distinguished between dATP and d2apTP and incorporated substantial amounts of d2apTP into template. Detergent-treated lymphocyte nuclei undergoing a high level of EBV DNA synthesis were shown to incorporate the 2-aminopurine analog of dATP into viral DNA. The relative inability of the EBV-induced DNA polymerase to distinguish between the two purine nucleotides reported here is consistent with previous reports on the ready incorporation of other nucleotide analogs into DNA polymerases induced by other herpesviruses. Because most antiherpes agents currently in use or under study are nucleotide analogs, the viral mutagenic properties of these drugs should be examined.

Characterization of the Epstein-Barr virion-associated DNA polymerase as isolated from superinfected and drug-stimulated cells.

We reported previously that Epstein-Barr (EB) virions and detergent-treated nucleocapsids co-purified with significant amounts of DNA polymerase activity that did not resemble other known host or viral polymerases. We report here that this species of DNA polymerase activity is present at early times after infection in lymphocytes abortively lytically infected (superinfected) with EB virus. However, studies with [35S]methionine labeling suggest de novo synthesis of enzyme has not occurred. Conversely, drug-stimulated lymphocytes that synthesize EB viral late proteins and virions contain this species of polymerase to the virtual exclusion of all others. This EB viral polymerase shows a marked preference for nicked and gapped double-stranded rather than primed single-stranded DNA templates. Its processiveness as measured on primed theta X174 phage DNA template is lower than that of lymphocyte beta polymerase. The data reported here are consistent with the hypothesis that the EB virion-associated DNA polymerase is synthesized at late times in the viral life cycle as are other structural proteins but it plays an important role early after viral infection. It is known that mature herpes virion DNA (including that of EB virus) is nicked and gapped and we propose that virion polymerase repairs the viral DNA at an early stage in infection before viral DNA replication begins.

Characterization of purified Epstein--Barr virus induced deoxyribonucleic acid polymerase: nucleotide turnover, processiveness, and phosphonoacetic acid sensitivity.

The Epstein--Barr (EB) virus induced DNA polymerase has been further purified and characterized with respect to nucleotide turnover activity, processiveness of synthesis, and interaction with phosphonoacetic acid (PAA). The polymerase as purified through denatured DNA--cellulose chromatography was inseparable from a labile nuclease activity associated with an equally labile DNA-dependent nucleotide turnover function. The EB virus induced DNA polymerase even in the absence of detectable nuclease or nucleotide turnover activity was less processive in its synthesis than were lymphocyte alpha polymerase or procaryotic polymerases, and this processiveness decreased with increasing purity of the enzyme. PAA was shown to inhibit nucleotide incorporation by the EB virus induced DNA polymerase in the presence of nuclease-activated native DNA template in the manner of a pyrophosphate analogue. Under conditions in which the concentration of 3'-hydroxyl termini in the template was more limited, PAA was not inhibitory. PAA likewise failed to significantly decrease the processiveness and the nucleotide turnover function of the polymerase.

Sedimentation characteristics of newly synthesized Epstein-Barr viral DNA in superinfected cells.

Replicating Epstein-Barr virus (EBV) DNA molecules isolated from superinfected Raji cells were shown to consist of 80S to 65S and 58S (mature) molecules Pulse-chase experiments showed that radioactive label of DNAS molecules with the larger sedimentation coefficients was partially chased into 58S labeled forms. Formation of large concatemers of viral DNA could not be detected at any time after superinfection. The continuous presence of the 65S viral DNA intermediate throughout the replicative cycle combined with the observed inhibition of EBV DNA synthesis by addition of nontoxic levels of ethidium bromide to the superinfected cell culture led us to propose that EBV replication proceeds via a relaxed circular DNA intermediate.

An endonuclease isolated from Epstein-Barr virus-producing human lymphoblastoid cells.

An endonuclease has been isolated from human B lymphoblastoid cells that copurifies with an exonucleolytic activity and has been shown to produce double-strand breaks and a high proportion of single-strandedness in phage lambda DNA in vitro. The data are consistent with a model in which single-strand cuts are made by the endonucleolytic activity, possibly in A+T-rich regions of the DNA, followed by creation of single-stranded regions (gaps) precessing from the site of a cut. Generation of overlapping gaps on opposite strands or of a gap opposite a nick would lead to the creation of the banding patterns that we have seen on electrophoretic gels. This endonucleolytic activity copurifies with other enzymes induced by Epstein-Barr virus that relate to the process of viral DNA replication in productively infected cells. However, a more general role is proposed for this class of eukaryotic endonuclease activities. A marked degree of single-strandedness has been found in the replicating DNAs of many eukaryotes, ad these gaps could be generated by endonucleases with associated exonucleolytic activity such as that reported here. This Epstein-Barr virus-induced nuclease activity has been shown to resemble the recBC nuclease isolated from the prokaryote Escherichia coli and also the endonuclease isolated from the eukaryote Chlamydomonas.

Deoxyribonuclease activity found in Epstein--Barr virus producing lymphoblastoid cells.

A deoxyribonuclease activity from Epstein--Barr (EB) virus producer lymphocyte cell lines which is correlated with viral production and which is not present in virus non-producer or negative lymphocyte cell lines has been purified 220-fold with 20% recovery and characterized. This nuclease copurifies through diethylaminoethylcellulose column chromatography with the EB virus induced deoxyribonucleic acid (DNA) polymerase in EB virus producer cells which was recently reported by this laboratory, but elutes as a separate peak of activity upon phosphocellulose chromatography. This nuclease activity has a sedimentation coefficient of 4.0 S, a strong divalent cation requirement, an alkaline pH optimum, and the ability to utilize both native and denatured lymphocyte DNA as substrate, reducing both to monophosphonucleosides.

Identification and partial purification of two EBV-associated DNA polymerases.

Virally induced DNA polymerases have been demonstrated in cells infected with a variety of herpesviruses. These include herpes simplex virus (Weissbach et al., 1973), Marek's disease virus (Boezi et al., 1974), equine herpesvirus (Allen et al., 1977), and cytomegalovirus (Huang, 1975a). Recently a new iododeoxyuridine (IUDR)-induced intracellular DNA polymerase in an Epstein-Barr virus (EBV)-producing hybrid cell line has also been reported (Miller et al., 1977). We present evidence that there are two different DNA polymerase activities associated with EBV, one intracellular and the other virion-associated. Both of these enzymes have certain biochemical characteristics which distinguish them from each other, and from the host-cell DNA polymerases found in lymphocytes.