Streamline density and lesion volume reveal a postero-anterior gradient of corpus callosum damage in multiple sclerosis.

BACKGROUND AND PURPOSE: Although interhemispheric disconnection significantly contributes to disability in multiple sclerosis (MS), the topography, timeline and relationship of callosal damage accrual with hemispheric damage are still unclear. METHODS: Streamline density and the presence of focal lesions in five callosal subregions were computed in 55 people with MS [13 relapsing-remitting (RRMS), 20 secondary progressive (SPMS), 22 primary progressive (PPMS)] and 24 healthy controls. RESULTS: Streamline density decrease was identified in SPMS in all corpus callosum (CC) subregions, in PPMS in the posterior CC and mid-posterior CC and in RRMS in the posterior CC. CC density was independently predicted by CC lesion volume and hemispheric lesion volume and independently predicted visuospatial memory, Expanded Disability Status Scale, manual dexterity and ambulation. CONCLUSIONS: The reduction in CC density across phenotypes suggests an earlier involvement of the posterior regions, followed only at a later stage by involvement of the anterior portions of the CC. Such interhemispheric disconnection seems to develop as a consequence of white matter macroscopic damage and exerts a relevant impact on motor and, to a lesser extent, cognitive disability.

Heart rate variability is reduced in acromegaly patients and improved by treatment with somatostatin analogues.

BACKGROUND: Cardiovascular complications, including arrhythmias and cardiac sudden death, are the most common causes of enhanced mortality in acromegaly. However, few data are available on cardiac autonomic functions and sympathovagal balance in acromegalic patients. OBJECTIVE: The aim of this study was to investigate both the time and frequency domain parameters of Heart Rate Variability (HRV), in order to characterize the cardiac autonomic functions in patients affected by acromegaly. This study correlated anthropometric, metabolic, echocardiographic parameters and blood pressure with those relating to HRV, to identify the main factors responsible for the HRV related alterations possibly present. We also aimed to analyze the effects of the treatment with somatostatin analogues (SSAs) on HRV. MATERIALS AND METHODS: This study enrolled 47 acromegalic patients (23 males, age 49.1 ± 13.5 years) and 37 (13 males) age matched (52.3 ± 13.3 years) healthy subjects. All participants underwent 12-lead 24 h ECG Holter recordings and a HRV analysis of the ECG tracings was performed. The parameters obtained from the time domain analysis of HRV included pNN50, SDNN, SDNN index, SDANN and RMSSD. The power spectral analysis of HRV was obtained by summing powers of the LF (low frequency) and the HF (high frequency) band. Sympathovagal balance was estimated by calculating the LF/HF ratio during 24 h and 15 min of clinostatism. The HRV of 28 acromegalic patients was studied before and after SSAs treatment. RESULTS: Acromegalic patients showed significantly lower SDNN and SDANN compared to controls. Diabetic and non-diabetic acromegalic patients showed decreased SDNN and SDANN, when compared to healthy subjects. Diabetic acromegalic patients had a lower LF/HF ratio during 24 h when compared to non-diabetic acromegalic patients. Similar results were obtained analyzing patients affected by acromegaly and impaired glucose tolerance. SDNN and SDANN were lowered by hypertension in the acromegalic population, when compared to controls, and hypertensive acromegalic patients also displayed a decreased LF/HF ratio during 24 h when compared to normotensive acromegalic subjects. Patients with ventricular arrhythmias in Lown classes 3-5 showed a decreased SDANN compared to patients in Lown class 0-2. The treatment with SSAs was able to ameliorate all the time domain parameters of HRV, without altering the 24 h LF/HF ratio. CONCLUSION: Cardiac autonomic functions and sympathovagal balance are altered in patients affected by acromegaly and could be ameliorated by SSAs therapy. HRV analysis allows an estimation of the autonomic sympathovagal balance and may be a useful clinical tool for the cardiac risk stratification in acromegalic patients.

Unusual vaginal angiomatous neoformation in a 3-year old pug.

A 3-year-old female pug (8 kg bodyweight) was referred by a local veterinarian for evaluation of vaginal bleeding. Historically, the owner indicated chronic haematic vaginal discharge initiated approximately 3 months ago, overall good appetite and slight general dullness. Two months prior to the visit, the bitch had been spayed by the local veterinarian, but vaginal bleeding persisted after surgery. Following a general examination and complete blood analysis, a genital tract examination was performed and, through vaginal endoscopy, a red, smooth, and apparently broad-based vaginal mass was found. An episiotomy was performed and on surgical exploration, the bleeding site was determined to be on the surface of a mass composed of two congested, adjacent cylindrical structures, symmetric to the vaginal sagittal midline, broad-based and quite regular on the surface. The mass appeared to be well delimited from the surrounding tissue by a thin capsula, and each cylindrical structure had a tributary vessel. Ectopic and dysplastic corpora cavernosa of the clitoris characterized by angiomatous proliferation, cavernous haemangioma and vascular hamartoma were considered in the differential diagnosis. The presence of small intralesional nerves identified by S100 immunostaining was used as a diagnostic clue to classify this unusual vaginal angiomatous neoformation as vascular hamartoma.

Pregnancy diagnosis in dairy cows by whey progesterone analysis: an ROC approach.

Concentration of progesterone in milk may be used to predict pregnancy status of dairy cattle by the 21st day after insemination. However, the accuracy of this method may be affected by fat-solubility of progesterone and sample storage conditions. After coagulation of a milk sample with rennet, an alternative method is to quantify progesterone concentration in whey with a novel, validated EIA. In this experiment, a receiver operating characteristic (ROC) analysis was performed to estimate the optimal discrimination point for whey progesterone concentration, using a sample of 991 Friesian cows evaluated between the 42nd and 44th day after insemination. Cows also were diagnosed for pregnancy by rectal palpation at this time. The overall conception rate at palpation was 57%. ROC analysis indicated that 259 pg/mL progesterone in whey was the most effective cutoff to discriminate correctly between pregnant and non-pregnant cows. Using this point for prediction, sensitivity was 98.2%, specificity was 70.9% and the area under ROC curve was 0.859, levels generally considered to denote moderate accuracy. The negative likelihood ratio at the cutoff of 259 pg/mL was 0.02, indicating satisfactory performance in detecting negative subjects, while the positive likelihood ratio (+LR=3.37) suggested average performance. In conclusion, EIA of progesterone concentration in whey is a viable method for predicting pregnancy status in cows. However, operators should take management objectives for the herd into account in determining the cutoff point and also considering important influencing variables such as conception rate in the herd. This method can provide diagnostic support for efforts to improve reproductive success, especially in low-fertility herds.

Oxytocin-induced PGF2alpha release in mares with and without post-breeding delayed uterine clearance.

The aim of the present study was to evaluate within 24 h post-ovulation oxytocin-induced PGF(2alpha) release in mares with and without post-breeding delayed uterine clearance (DUC). Twenty-one of 34 mares with a variable amount of intrauterine fluids accumulation were considered to be affected by delayed uterine clearance (DUC group), while the other 13 mares did not show any uterine fluid accumulation, and were considered as controls (WDUC group). Both DUC and WDUC mares were administered with 20 IU oxytocin i.m. 90 min after the ultrasound examination performed 24 h after breeding. Immediately before, 5 and 10 min after oxytocin administration, blood samples were collected for 15-ketodihydro-PGF(2alpha) (PG-metabolite), 17beta-estradiol, and progesterone analysis. Ultrasonography performed 24 h after oxytocin treatment showed a complete uterine clearance in all DUC mares. The oxytocin-induced PG-metabolite increase was detected in 71.4% DUC mares compared with 38.5% in WDUC group, with a positive trend of release, as evidenced from 5 min after oxytocin administration. In WDUC mares, no significant differences in oxytocin-induced PG-metabolite trend of release were observed. In conclusion, the results of the present study showed the importance of PGF(2alpha) involvement in the pathogenesis of post-breeding DUC in the mare.

Comparison between cloprostenol-induced and spontaneous oestrus fertility in dairy cows.

A short calving to conception interval is of main importance to achieve high economic efficiency in dairy cow industry. In order to reduce this interval, several hormonal treatments have been put on the market, in which cloprostenol, a synthetic analogue of prostaglandin F2alpha (PGF2alpha). The aim of this study was to compare fertility of cloprostenol-induced oestrus to that of spontaneous oestrus in dairy cows. In a group of 525 cows, 280 (treated group) were administered 0.5 mg cloprostenol i.m. after transrectal corpus luteum (CL) detection, and inseminated at detected oestrus during the following week. The other 245 cows (control group) were inseminated during spontaneous oestrus. Whey progesterone concentrations were checked at treatment and at insemination in order to remove from the study cows whose P4 levels indicate a non-functional CL, or a lack of luteolysis respectively. Moreover, cows that were not inseminated due to genital problems were also excluded from this study. Conception (59% vs 54.5%) and calving rates (93.7% vs 93%) were not significantly different between the two groups.

Plasma concentrations of 15-ketodihydro-PGF(2alpha), cortisol and progesterone during manual twin reduction in thoroughbred mares.

The aim of the present study was to highlight the effect of two different techniques of one embryo crushing on some hormonal changes. Ten twinning mares were submitted to the mobile or fixed manual crushing of one blastocyst within day 19 after the last mating. Blood sample was collected from 20 min before to 90 min, 24 and 72 h after the procedure was performed to analyse 15-ketodihydro-PGF(2alpha), cortisol and progesterone plasma concentrations. Singleton pregnancy diagnosis was checked 72 h after crushing and at term of pregnancy. Because the unwanted crushing of both embryos occurred in one mare during the attempt of manual separation of the twins, that mare was not included in the evaluation of crushing-induced hormonal changes. No significant differences in hormonal concentrations were observed after one embryo crushing and also when the effect of the mobile (n = 6) or fixed (n = 3) technique was specifically evaluated. When the effect of the two techniques on each post-crushing sampling time hormonal levels was analysed, only a higher cortisol level 30 min after the fixed compared with the mobile technique was observed. The crushing performed within 19 days of gestation does not induce significant changes in 15-ketodihydro-PGF(2alpha), cortisol or progesterone plasma concentrations. When the fixed technique was performed, only a temporary higher cortisol concentration was seen 30 min after crushing, suggesting that the fixed technique might be responsible for a slight level of stress for the mare.

Relationship between pharmacological induction of estrous and/or ovulation and twin pregnancy in the Thoroughbred mares.

The aim of this study was to evaluate the possible relationship of pharmacological induction of estrous and/or ovulation with the occurrence of twin pregnancies in Thoroughbred mares. Out of 680 mares, 356 received one of the following treatments during the estrous cycle in which they became pregnant: injection of 0.5mg of cloprostenol at the ultrasonographic detection of a CL (n=86); injection of 5000 IU human chorionic gonadotropin (hCG) immediately before mating (n=221); injection of 0.5mg of cloprostenol at the ultrasonographic detection of a CL plus injection of 5000 IU hCG immediately before mating on cloprostenol-induced estrous (n=49). The other 324 mares, not treated for induction of estrous or ovulation in the estrous cycle resulting in pregnancy, were used as control group. The occurrence of twin and single pregnancies in treated and control mares underlines that the percentage of twin pregnancy in treated mares (16.6%) was statistically significantly higher (P<0.0001; odds ratio, OR=2.87) than the percentage of twinning in the control group (6.5%). Comparison of the occurrence of twins between treatments revealed a statistically significant difference between mares treated with hCG alone compared to animals given prostaglandin F2alpha (PGF2alpha) plus hCG. The results show a statistically significant difference for each treatment compared to controls, with the least difference (P<0.05; OR=2.18) for the comparison between hCG treatment group and controls, a significance of P<0.01; OR=3.05 for the comparison between PGF2alpha treatment and controls, and a highly statistically significant difference (P<0.0001; OR=6.37) for the comparison between PGF2alpha plus hCG-treated animals and controls.

17beta-estradiol, progesterone and testosterone concentrations in cystic fluids and response to GnRH treatment after emptying of ovarian cysts in dairy cows.

The aim of this study was to determine possible links between steroidogenic activity of single ovarian cysts and response to intramuscular treatment with 20 microg of buserelin (GnRH-analogue) after cyst emptying, in pluriparous Friesian cows bearing a singleton cyst treated not earlier than 55 days post-partum. Progesterone, 17beta-estradiol and testosterone were determined in cystic fluids collected by needle aspiration of the cyst. Of the cows, 75.6% began ovarian cyclicity within 30 days after treatment with a conception rate of 64.7%. In this study it was found that as progesterone concentration in cystic fluids rose, the number of positive responses to the treatment fell.

Ultrasonographic appearance of tissue is a better indicator of CL function than CL diameter measurement in dairy cows.

In this study, the ovaries of 99 randomly selected Friesian cows were examined by ultrasonography measuring the diameter and evaluating the appearance of corpora lutea (CLs) in order to assess the most reliable method for their functional classification. Concurrently, blood samples were taken and analyzed for plasma progesterone (P4) concentration. On the basis of the ultrasonographic measurement of the diameter of the CL, three groups were established: (A) CL not detected (n = 30), (B) CL psi < 20 mm (n = 22), and (C) CL psi > or = 20mm (n = 47). On the basis of the ultrasonographic appearance, three different groups were established: (A) CL not detected (n = 30), (B) evolving CL (n = 25), and (C) mid-cycle CL (n = 44). On the basis of the P4 values, CLs were functionally classified in the following three groups: (A) CL not detected when plasma P4 was lower than 1 ng/ml (n = 27), (B) evolving CL when plasma P4 was between 1 and 4 ng/ml inclusive (n = 29), and (C) mid-cycle CL when plasma P4 was more than 4 ng/ml (n = 43). The degree of agreement between plasma P4 concentrations and either ultrasonographic classification (diameter or appearance) was highly significant (P < 0.001). However, the results of the present study suggest that for the evaluation of functional classification of the CL in cows ultrasonographic appearance is more reliable than the evaluation of the diameter.

Correlations among body temperature, plasma progesterone, cortisol and prostaglandin F2alpha of the periparturient bitch.

The results of this study suggest that, besides the irrelevant role of body temperature measurement to predict the impending parturition in the bitch, progesterone and 15-ketodihydroprostaglandin F2alpha plasma level records could be more suitable to detect the approaching whelping in this species. More interesting was the statistically significant substantial increase in body temperature beginning 12 h after the onset of parturition. Therefore, if any significant increase in body temperature is recorded at the end of pregnancy without the beginning of the expulsion of fetuses, it could indicate problems at parturition. In this study, cortisol levels increased significantly at the time of delivery and remained high 12 h after the beginning of parturition, decreasing within 36 h after the onset of whelping. 15-ketodihydro-prostaglandin F2alpha levels increased significantly 24 h before parturition and again at the onset of whelping. Progesterone levels decreased significantly, starting 24 h before the onset of whelping and remained low after delivery.

Agreement between ultrasonographic classification of the CL and plasma progesterone concentration in dairy cows.

We evaluated the agreement between ultrasonographic characteristics of the corpus luteum (CL) and plasma progesterone (P4) concentration in dairy cows. In Phase I of the study, the ovaries of 8 cows were ultrasonographically examined, and P4 was analyzed daily from estrus (Day 0) to Day 4, then at Day 7 and Day 10, and again daily from Day 17 to the onset of next estrus. In Phase 2, the ovaries of 157 randomly selected Friesian cows were examined once by ultrasonography, and blood samples collected concurrently were analyzed for plasma P4. On the basis of the P4 values, the function of CLs was classified as follows: 1) non-secretory CL when plasma P4 was lower than 1 ng/mL (n=41); 2) evolving CL when plasma P4 was between 1 and 4 ng/mL (n=55); and 3) mid-cycle CL when plasma P4 was more than 4 ng/mL (n=61). On the basis of ultrasonographic examination, 3 additional groups were established (absence of CL, evolving CL, midcycle CL). Ultrasonographic characteristics and size of Day 3 to 4 CLs and their respective plasma P4 concentrations were not distinguishable from those of CLs observed 3 to 4 d before the subsequent estrus. The degree of agreement between the two classification was 72%. The data indicate that the functional classification of CLs is difficult to determine based on ultrasonography alone.

Effects of slow and ultrarapid freezing on morphology and resumption of meiosis in immature cat oocytes.

This study examined the ability of immature cat oocytes to survive after cryopreservation by evaluating their subsequent development following maturation in vitro. The effect of slow and ultrarapid freezing using one of two cryoprotectants dimethylsulphoxide (DMSO) or 1,2-propanediol (PROH) at different concentrations (1.5 or 3.0 mol-1) and the slow freezing with the cryoprotectant ethylene glycol (EG 1.5 mol l-1 and 3.0 mol l-1) were tested. Morphology, resumption of meiosis and metaphase II rates of oocytes were recorded after thawing. Freshly collected oocytes were used as controls. Results indicate that immature cat oocytes can survive, resume meiosis and achieve metaphase II in vitro after freezing. The highest rates of resumption of meiosis and metaphase II were achieved with slow freezing and 1.5 mol DMSO or EG l-1 (DMSO: 47.4%, 18/38 and 23.7%, 9/38 and EG: 52%, 13/25 and 20%, 5/25, respectively). The ultrarapid procedure did not result in resumption of meiosis in vitro, despite intact morphology of the oocytes after thawing. These results suggest that morphology of oocytes after freezing and thawing has no predictive value for their ability to resume meiosis.

Flow cytometric analysis of cellular DNA content in paraffin wax-embedded specimens of canine mammary tumours.

Flow cytometric analysis of DNA content was performed on 72 formalin-fixed, paraffin wax-embedded samples of canine mammary tumours. DNA content was correlated with histological appearance, stage of the tumour, and survival after mastectomy. All benign tumours (14 cases) were diploid. Amongst the malignant tumours, a higher incidence of aneuploid tumours (50 per cent) was observed in cases with lymph nodal metastases in comparison with those without metastases (26 per cent). All dogs with benign tumours survived for at least two years after mastectomy and so did 26 (81 per cent) of 32 DNA diploid malignant cases without metastases. A 67 per cent (eight of 12 cases) survival rate was found in the dogs with DNA aneuploid malignant tumours without metastases, while a very low survival rate (17 per cent) was observed in the group with malignant tumours with metastases, independently of the extent of ploidy.