Treatment of full thickness focal cartilage lesions with a metallic resurfacing implant in a sheep animal model, 1 year evaluation.

BACKGROUND: Full depth focal cartilage lesions do not heal spontaneously and while some of these lesions are asymptomatic they might progress to osteoarthritis. Treatment for these lesions is warranted and the gold standard treatment at younger age remains biological healing by cell stimulation. In the middle-age patient the success rate of biologic treatment varies, hence the surge of non-biological alternatives. Our objective was to evaluate the efficacy and safety of a metallic implant for treatment of these lesions with respect to the long-term panarticular cartilage homeostasis. METHODS: The medial femoral condyle of 16 sheep was operated unilaterally. A metallic implant was inserted in the weight-bearing surface at an aimed height of 0.5 mm recessed. Euthanasia was performed at 6 or 12 months. Implant height and tilt was analyzed using a laser-scanning device. Damage to cartilage surfaces was evaluated macroscopically and microscopically according to the Osteoarthritis Research Society International (OARSI) recommendations. RESULTS: Thirteen sheep were available for evaluation and showed a varying degree of cartilage damage linearly increasing with age. Cartilage damage of the medial tibial plateau opposing the implant was increased compared to the non-operated knee by 1.77 units (p = 0.041; 95% CI: 0.08, 3.45) on a 0-27 unit scale. Remaining joint compartments were unaffected. Implant position averaged 0.54 recessed (95% CI: 0.41, 0.67). CONCLUSIONS: Our results showed a consistent and accurate placement of these implants at a defined zone. At this position cartilage wear of opposing and surrounding joint cartilage is limited. Thus expanded animal and human studies are motivated.

Fixation of a double-coated titanium-hydroxyapatite focal knee resurfacing implant: a 12-month study in sheep.

OBJECTIVE: Focal cartilage lesions according to International Cartilage Repair Society (ICRS) grade 3-4 in the medial femoral condyle may progress to osteoarthritis. When treating such focal lesions with metallic implants a sound fixation to the underlying bone is mandatory. We developed a monobloc unipolar cobalt-chrome (Co-Cr) implant with a double coating; first a layer of commercially pure titanium (c.p.Ti) on top of which a layer of hydroxyapatite (HA) was applied. We hypothesised that such a double coating would provide long-lasting and adequate osseointegration. DESIGN (MATERIALS AND METHODS): Unilateral medial femoral condyles of 10 sheep were operated. The implants were inserted in the weight-bearing surface and immediate weight-bearing was allowed. Euthanasia was performed at 6 (three animals) or 12 months (six animals). Osseointegration was analysed with micro-computer tomography (CT), light microscopy and histomorphometric analyses using backscatter scanning electron microscopy (B-SEM) technique. RESULTS: At 6 months one specimen out of three showed small osteolytic areas at the hat and at 12 months two specimens out of six showed small osteolytic areas at the hat, no osteolytical areas were seen around the peg at any time point. At both time points, a high total bone-to-implant contact was measured with a mean (95% confidence interval - CI) of 90.6 (79-102) at 6 months and 92.3 (89-95) at 12 months, respectively. CONCLUSIONS: A double coating (Ti + HA) of a focal knee resurfacing Co-Cr implant was presented in a sheep animal model. A firm and consistent bond to bone under weight-bearing conditions was shown up to 1 year.

Focal knee resurfacing and effects of surgical precision on opposing cartilage. A pilot study on 12 sheep.

BACKGROUND: Full thickness cartilage lesions (ICRS grade 3-4) and focal lesions of degenerative origin may progress to osteoarthritis (OA). Such focal lesions can be treated by metallic implants. We hypothesized that such treatment results in opposing surface cartilage damage that correlates with implant position (height) relative to the adjacent cartilage surface. This relationship was investigated using a sheep animal model. METHODS: Both medial femoral condyles of 12 sheep were operated. The implants, were inserted in the weight-bearing surface at different heights relative to the surrounding cartilage. Euthanasia was performed at 6 or 12 weeks. After retrieval, implant height was analyzed using laser scanning. Damage to the opposing tibial cartilage was evaluated macroscopically and microscopically according to the modified Mankin score. RESULTS: Twenty-two knees were available for evaluation and showed cartilage lesions ranging from severe damage (Mankin stage 11) to almost pristine conditions (Mankin stage 1). There was a strong correlation between implant height and cartilage damage. Standard deviation from the aimed implant height was 0.47 mm. CONCLUSIONS: Our results showed significant surgical imprecision and protruding implants imposed severe cartilage damage. We therefore suggest implants should be placed recessed (approx. 0.5 mm) below the surrounding cartilage in this animal model. These results encourage further studies of metallic implants yet the utmost precision regarding position is required.

Postconditioning and remote postconditioning of ischemic rat cardiac grafts.

BACKGROUND: Global warm ischemia of transplanted cardiac grafts is associated with apoptosis and subsequent tissue necrosis. It is suggested that postconditioning (PostC) may ameliorate the early outcome of cardiac grafts after ischemia-reperfusion. We investigated whether PostC and remote postconditioning (RPostC) have a histopathologic impact after transplantation of warm ischemic rat cardiac grafts. MATERIALS AND METHODS: Thirteen rats were randomized to undergo either PostC or RPostC after heterotopic transplantation of ischemic cardiac grafts. Six rats without intervention besides transplantation served as controls (control). The recipient rats were sacrificed 24 h after transplantation to evaluate histopathology and apoptotic index. RESULTS: No significant differences were observed between the study groups in terms of graft heat shock protein 70 and oxygen radical absorbing capacity, an indicator of antioxidant capacity. While apoptosis and PCARB, a marker of protein oxidation and oxidative stress, decreased after RPostC (p < 0.05), contraction band necrosis was less prominent in both PostC and RPostC. CONCLUSION: Both PostC and RPostC have a histopathologic impact after 24 h of reperfusion of warm ischemic rat cardiac grafts.

Diamniotic omphalopagus conjoined twins in a diamniotic pregnancy.

OBJECTIVE: To present antepartal diagnostic findings including 2D/3D ultrasonography in a rare case of conjoined twins with diamniotic placentation. METHODS: In ultrasonographic examination, a communicating structure containing solid and cystic components together with cord vessels was found between the fetuses. The long axes of the fetuses were not parallel and they moved independently. Bladder configurations were absent. A fetal membrane from the chorioidal plate was imaged. RESULTS: Postmortem pathological examination revealed that the twins were conjoined by way of fused cloacal exstrophy and omphalocele, and the separate amniotic cavities communicated via a fused allantoic cavity. The placenta was monochorionic and diamniotic. DNA analysis of the twins and the placenta confirmed the monozygotic origin. CONCLUSION: Visualization of the amniotic membrane does not rule out conjoined twins in rare cases of monochorionic twin pregnancies.

Effect of freezing on sensory quality, shear force and water loss in beef M. longissimus dorsi.

The objective of this study was to determine how sensory quality, shear force and water loss differ between beef stored either chilled or frozen before cooking. Meat tenderness was analysed instrumentally and sensorially using both a consumer panel and a semi-trained panel. Both M. longissimus dorsi (LD) from eight young Holstein bulls were cut into eight samples, weighed, vacuum packed and aged at 4°C for 2, 7 or 14 days. After ageing, the frozen samples were kept at -20°C prior to heat treatment. Water holding capacity was recorded as purge or thawing loss and cooking loss or as combined loss. Sensory analyses were performed on samples aged 7 days. Peak force values declined with ageing time and freezing. Frozen meat aged 2 days had the same peak force values as chilled meat aged 7 days. Total energy was the same for both treatments at day 2 and 7, whereas at day 14 frozen samples showed significantly higher values than chilled samples. The sensory panel experienced the chilled meat to be more tender, juicier and having a more intense meat taste than the frozen meat, whereas the consumers could not find any significant difference in degree of liking. Water holding capacity was lower for the frozen samples. The results indicate that conclusions from studies concerning sensory quality of beef will depend on whether the meat has been kept chilled or frozen before testing.

Catestatin, vasostatin, cortisol, and visual analog scale scoring for stress assessment in healthy dogs.

The neuroendocrine glycoprotein chromogranin A is a useful biomarker for stress in humans. Chromogranin A epitopes catestatin and vasostatin can be measured in dogs using radioimmunoassays. The objective of this study was to evaluate catestatin and vasostatin as canine stress biomarkers in a clinical setting. Blood and saliva were collected from 33 healthy dogs that were familiar with sampling procedures and the animal hospital environment (control group) and 30 healthy dogs that were unacquainted (stress group). During sampling, stress behavior was scored by the same observer using visual analog scale (VAS). Plasma was analyzed for catestatin and vasostatin, serum for cortisol, and saliva for catestatin. Differences between groups were analyzed using two-sample t-tests and P<0.05 was considered significant. Stress behavior VAS score in the control group was significantly lower than in the stress group during blood (P=0.002) and saliva (P=0.0009) sampling. Serum cortisol and saliva catestatin concentrations in the stress group were higher than the control group (P=0.003 and P<0.0001, respectively). Serum cortisol concentrations were correlated with those of saliva (r=0.34, P=0.04) and plasma catestatin (r=0.29, P=0.03). Plasma catestatin and vasostatin did not differ significantly between groups. In conclusion, concentrations of saliva catestatin, and serum cortisol, and stress behavior VAS scores were significantly higher in the stress group. The results indicate that saliva catestatin may be useful as a biomarker for acute psychological stress in dogs.

Differentiation between pyometra and cystic endometrial hyperplasia/mucometra in bitches by prostaglandin F2alpha metabolite analysis.

Bitches with pyometra are potential emergency cases which may be clinically difficult to differentiate from cases of cystic endometrial hyperplasia (CEH) in combination with mucometra. In the present study plasma prostaglandin F(2alpha), as measured by its main metabolite 15-keto-13,14-dihydro-PGF(2alpha) (PG-metabolite) concentrations, blood biochemical and hematological parameters were measured in 59 bitches with pyometra, 10 bitches with CEH and nine controls to determine if PG-metabolite could differentiate between the three uterine conditions. Bitches with pyometra had significantly higher plasma levels of PG-metabolite than bitches with CEH (P=0.002) and the controls (P=0.002). PG-metabolite analysis alone had a high sensitivity (98.3%) and a high specificity (80.0%) for the differentiation of pyometra versus CEH in bitches where fluid in the uterus was diagnosed. When a combination of PG-metabolite and percentage band neutrophils (PBN) was used for differentiation of the two diagnoses, a sensitivity of 100% and specificity of 90.0% was obtained. This means that the combination of PG-metabolite and PBN analysis allows for differentiation between cases of pyometra and CEH. If the PG-metabolite level in a bitch is >or=4,524 pmol l(-1), there is a 99% probability of the diagnosis pyometra versus CEH. Levels of PG-metabolite >or=3,054 pmol l(-1), >or=2,388 pmol l(-1) or>or=1,666 pmol l(-1) indicates a 95%, 90% or 80% probability of pyometra, respectively. At high PG-metabolite levels (above about 3,000 pmol l(-1)), PG-metabolite alone is enough for differentiation of pyometra versus CEH. The results of the present study showed that PG-metabolite analysis is valuable in the diagnosis and prediction of severity of uterine diseases.

Pyometra in bitches induces elevated plasma endotoxin and prostaglandin F2alpha metabolite levels.

Endotoxemia in bitches with pyometra can cause severe systemic effects directly or via the release of inflammatory mediators. Plasma endotoxin concentrations were measured in ten bitches suffering from pyometra with moderately to severely deteriorated general condition, and in nine bitches admitted to surgery for non-infectious reasons. Endotoxin samples were taken on five occasions before, during and after surgery. In addition, urine and uterine bacteriology was performed and hematological, blood biochemical parameters, prostaglandin F2alpha metabolite 15-ketodihydro-PGF2alpha (PG-metabolite), progesterone and oestradiol (E2-17beta) levels were analysed. The results confirm significantly increased plasma levels of endotoxin in bitches with pyometra and support previous reports of endotoxin involvement in the pathogenesis of the disease. Plasma concentrations of PG-metabolite were elevated in pyometra bitches and provide a good indicator of endotoxin release since the concentrations were significantly correlated to the endotoxin levels and many other hematological and chemistry parameters. The gamma-globulin serum protein electrophoresis fraction and analysis of PG-metabolite can be valuable in the diagnosis of endotoxin involvement if a reliable, rapid and cost-effective test for PG-metabolite analysis becomes readily available in the future. Treatment inhibiting prostaglandin biosynthesis and related compounds could be beneficial for bitches suffering from pyometra.

Primary uterine inertia in 27 bitches: aetiology and treatment.

OBJECTIVES: To evaluate two treatment methods in bitches with primary uterine inertia in relation to blood concentrations of oxytocin, calcium and glucose. METHODS: A prospective study where 27 bitches with primary inertia were randomly allocated to treatment groups I and II. Blood samples were collected before and after treatment. Group I was treated with a combination of intravenous calcium solutions and oxytocin and group II with oxytocin only. If unsuccessful, caesarean sections or additional medical treatment was pursued. RESULTS: Plasma oxytocin concentrations were 35+/-15 pmol/l in group I and 30+/-15 pmol/l in group II. Before treatment, the total serum calcium concentrations were 2.1+/-0.2 mmol/l in both the groups, and blood glucose values were 7.0+/-0.5 mmol/l (group I) and 7.3+/-1.4 mmol/l (group II). The number of bitches subjected to caesarean sections and the number of puppies born did not differ between the two groups. CLINICAL SIGNIFICANCE: This study indicates that low plasma oxytocin levels is a cause of primary inertia in bitches with normal serum calcium concentrations and aggravates the condition in bitches with low calcium levels. Hypoglycaemia was not observed. The outcome of labour did not differ between groups I and II.

Immunological variation and immunohistochemical localization of HIV-1 Nef demonstrated with monoclonal antibodies.

OBJECTIVE: To study the immunological and immunohistochemical nature of HIV-1 Nef. DESIGN: Monoclonal anti-Nef antibodies were generated and used to identify antigenic epitopes in Nef, to study immunological cross-reactivity between Nef from different isolates and to reveal the subcellular localization of Nef. METHODS: Monoclonal antibodies against recombinant HIV-1 Nef protein (BRU isolate) were generated in BALB/c mice. The epitope mapping was carried out with the use of overlapping 15-20mer lipopeptides linked to a lipid group at the amino-terminus. Immunoperoxidase method was used for histochemical studies. RESULTS: Ten stable antibody-producing clones, mainly of the immunoglobulin (Ig) G1 subtype, with strong Western blot and enzyme-linked immunosorbent assay reactivity toward the recombinant Nef protein, were obtained. The epitopes recognized were located on amino-acid sequences 21-41, 31-50, 51-71, 61-80, 151-170, 161-180, and 171-190. All 10 monoclonal antibodies also reacted with the native Nef of HIV-1BRU, and eight reacted with native HIV-1IIIB. Most antibodies also reacted with Nef from more divergent HIV-1 strains. In Western blotting, two forms of Nef (24 and 27 kDa) were observed with most isolates studied. Immunohistochemical staining of HIV-1-infected H9 or MT-4 lymphoid cells demonstrated that Nef was expressed mainly in the Golgi complex and at the nuclear membrane, but occasionally also in the nucleus. The nuclear localization of Nef was especially frequent in the HIV-1-infected MT-4 cells. CONCLUSIONS: Our findings suggest that Nef is expressed in two isomorphic forms, and that it may also act as a nuclear protein and thus have a direct regulatory function at the RNA/DNA level.

HLA-Dw1 and BfF as protective markers in multiple sclerosis.

The frequencies of Properdin factor B allotypes were studied in 54 multiple sclerosis patients and 58 healthy control subjects, and the association of various phenotypes with HLA-Dw1 and Dw2 antigens (found with decreased and increased frequency in MS patients, respectively) was further studied. SS genotype was found in 76% of MS patients and 63% of control subjects (n.s.) F types (FS + FF) were found to be strongly associated with HLA-Dw1 in control subjects (P less than 0.0014), whereas neither SS nor F containing types were associated with Dw2. The findings support the concept of a hypothetical MS resistance factor in the HLA gene area.

Histocompatibility antigens in familial multiple sclerosis in a high-risk area of the disease.

Fourteen Finnish families, each with two or more cases of multiple sclerosis (MS), were analysed for HLA antigens. Seventy family members were studied. They were all born in a high-risk area of MS and came from families with an unusually high frequency of MS. The frequencies of B7, B12 and DR2 were increased both in patients and their healthy relatives, whereas A1, B8 and DR1 were decreased in patients. MS patients shared the HLA haplotypes more often than expected.

Biochemical phenotypes of Escherichia coli in dogs: comparison of isolates isolated from bitches suffering from pyometra and urinary tract infection with isolates from faeces of healthy dogs.

Escherichia coli from dogs suffering from pyometra and/or urinary tract infection (UTI) were compared with faecal E. coli. We studied 79 isolates from pyometra, 96 from UTI, 91 from faeces of healthy dogs and 60 from faeces of bitches with pyometra. The isolates included 16 bitches with pyometra where E. coli were isolated from both the uterus and the urinary tracts. All isolates were typed with the aid of an automated typing system for biochemical fingerprinting (the PhP-EC system). The PhP types of uterine E. coli isolates from bitches with pyometra showed higher homogeneity, calculated as the mean of similarity between isolates (rmean = 0.79) than epidemiologically unrelated isolates, such as those isolated from faeces (rmean = 0.69) or UTI (rmean = 0.70). This finding suggests that many of the pyometra isolates derives from related clones. Mucoid and nonmucoid uterine isolates from the same sample were generally of the same PhP type. In all 10 cases uterine E. coli isolates were identical or very similar to isolates from faeces of the same bitch. Furthermore, E. coli isolates from both UTI and the infected uterus in the same bitch were identical or very similar in 88% of the cases. It was concluded that E. coli associated with canine pyometra derive from the faecal flora and that the urinary tract is infected by the same E. coli clone as the uterus of a bitch with pyometra.

Electrochemical treatment (EChT) effects in rat mammary and liver tissue. In vivo optimizing of a dose-planning model for EChT of tumours.

BACKGROUND: A reinvented technique for tumour therapy, electrochemical treatment (EChT), is attracting increasing attention. This study compared results from treatment of liver and mammary tissue focusing on destruction and pH changes in the tissue close to the treatment electrodes. Subsequently, data were compared with a dose-planning model. METHODS: Mammary or liver tissue in 50 adult female Sprague Dawley rats was given EChT with a constant, direct current. The electrodes used were Pt/Ir (9:1) with spherical tips. In situ pH measurements were taken with a micro-combination glass electrode. RESULTS: Spherical lesions were produced in both liver and mammary tissue. No significant difference was detected when comparing the size of the lesions in the two kinds of tissue. Similar pH profiles were obtained in tissue surrounding the electrodes, with pH values changing rapidly from unphysiological to neutral status within the space of a few millimetres. The pH at the border of the macroscopic destruction zone, regardless of tissue type or coulomb dosage, correlated well with specific values (4.5-5.5 at the anode and between 9 and 10 at the cathode). CONCLUSION: The analogous destruction patterns in mammary and liver tissue support the hypothesis that EChT has similar results in at least these two different types of tissue. This implies that the destructive pattern caused by the treatment may be the same also in tumours.

Electrochemical treatment of tumours.

The electrochemical treatment (EChT) of tumours implies that tumour tissue is treated with a continuous direct current through two or more electrodes placed in or near the tumour. The treatment offers considerable promise of a safe, simple and relatively noninvasive anti-tumour therapy for treatment of localised malignant as well as benign tumours. Although more than 10,000 patients have been treated in China during the past 10 years, EChT has not yet been universally accepted. The reason for this is the lack of essential preclinical studies and controlled clinical trials. Uncertainties regarding the destruction mechanism of EChT also hinder the development of an optimised and reliable dose-planning methodology. This article reviews the collected Chinese and occidental experiences of the electrochemical treatment of tumours, alone and in combination with other therapies. The current knowledge of the destruction mechanism underlying EChT is presented along with different approaches towards a dose planning methodology. In addition, we discuss our view of different important parameters that have to be accounted for, if clinical trials are to be initiated outside of China.

Breed risk of pyometra in insured dogs in Sweden.

An animal insurance database containing data on over 200,000 dogs was used to study the occurrence of pyometra with respect to breed and age during 1995 and 1996 in Swedish bitches <10 years of age. A total of 1,803 females in 1995 and 1,754 females in 1996 had claims submitted because of pyometra. Thirty breeds with at least 800 bitches insured each year were studied using univariate and multivariate methods. The crude 12-month risk of pyometra for females <10 years of age was 2.0% (95% confidence interval = 1.9-2.1%) in 1995 and 1.9% (1.8-2.0%) in 1996. The occurrence of pyometra differed with age, breed, and geographic location. The risk of developing pyometra was increased (identified using multivariate models) in rough Collies, Rottweilers, Cavalier King Charles Spaniels, Golden Retrievers, Bernese Mountain Dogs, and English Cocker Spaniels compared with baseline (all other breeds, including mixed breed dogs). Breeds with a low risk of developing the disease were Drevers, German Shepherd Dogs, Miniature Dachshunds, Dachshunds (normal size), and Swedish Hounds. Survival rates indicate that on average 23-24% of the bitches in the databases will have experienced pyometra by 10 years of age. In the studied breeds, this proportion ranged between 10 and 54%. Pyometra is a clinically relevant problem in intact bitches, and differences related to breed and age should be taken into account in studies of this disease.

Wound healing and soft tissue effects of CO2, contact Nd: YAG and combined CO2-Nd: YAG laser beams on rabbit trachea.

Rabbit trachea was used as an experimental model to study tissue effects and healing of full-thickness tracheal lesions produced by CO2, contact Nd: YAG and combined, coaxial CO2-Nd: YAG (Combo) laser beams. Two power settings (10 W and 16 W) were used with CO2 and contact Nd: YAG lasers. Three different CO2/Nd:YAG power ratios (1:1, 1:2 and 1:4) and power settings (12 W 15 W and 16 W) were used with the Combolaser. Histological specimens for light and transmission electron microscopy were prepared immediately and 1, 3, 5, 7, 14 and 21 days postoperatively. The wound with the most precise and fastest healing was produced by contact Nd: YAG laser. CO2 laser produced a moderate amount of charring and the largest amount of coagulated tissue with a slightly prolonged healing period. In the acute phase, tissue defects produced by the Combolaser with power ratios 1:1 and 1:2 resembled the CO2 laser lesions but with slightly less charring. The power ratio 1:4 diminished the cutting properties of the beam considerably. During the healing period the Combolaser produced the most intensive inflammation and granulation tissue formation resulting in delayed regeneration of the lesion. In transmission electron micrographs the most severe damage to chondrocytes was seen after using the Combolaser. These findings indicate that the Combolaser produces deeper tissue damage than CO2 or contact Nd:YAG laser. However, the Combolaser appears to be suitable for tracheobronchial operations, owing to its good simultaneous cutting and haemostatic properties.

Evaluation of possible spinal neurotoxicity of clonidine.

An experimental investigation was carried out concerning the potential neurotoxic effects of clonidine. For this purpose intrathecal or extradural catheters were implanted in dogs. Clonidine at a dose level of 25 micrograms/kg or 12.5 micrograms/kg, or placebo, was then administrated via the catheter once daily for 14 consecutive days. The spinal cord and the nerve roots were then taken for neuropathological analysis. Observation of the neurological behaviour of the animals and the results of the morphological investigation support the conclusion that clonidine under these circumstances does not have any significant neurotoxic effects.

Change of diurnal heart rate patterns during pregnancy and lactation in dogs (Canis familiaris).

Pregnancy and lactation involve great demands on the cardiovascular system. The purpose of this study was to investigate how the heart rate and diurnal heart rate pattern change when dogs become pregnant or lactate. Five clinically healthy female beagle dogs were mated, and delivered three to seven healthy puppies. The heart rate was investigated with 24-h ECG (Holter) once during anoestrus, at 3, 5, 7 and 9 weeks of pregnancy, and at week 4 postpartum (lactation). However, at 9 weeks, the ECG could not be recorded for the fully 24 h in 4 of 5 dogs, because labour started and the dogs then appeared disturbed by the recordings. The results at this date are not included in the statistical comparison. The heart rate increased progressively during pregnancy and was still elevated at 4 weeks of lactation. During late pregnancy the difference in heart rates between daytime and nighttime became smaller, but the heart rate was significantly higher in daytime in all periods. In conclusion, the increased heart rates during pregnancy and lactation reflect increased demands on the cardiovascular system and may be important to consider in clinical practice.