Analysis of genomic mutation and immunohistochemistry of platelet‐derived growth factor receptors in canine vascular tumours

We examined whether mutation of the platelet‐derived growth factor receptor protein tyrosine kinase (PDGFR)‐α and PDGFR‐β genes contributes to their overexpression in canine vascular tumours. Genomic sequences of trans‐ or juxtamembrane regions of PDGFR‐α and PDGFR‐β were analysed with immunohistochemical staining and polymerase chain reaction‐direct sequencing using DNA from paraffin‐embedded neoplastic tissues of 27 hemangiosarcomas (HSAs) and 20 hemangiomas (HAs). Immunohistochemically, 75% of the HA cases were positive for PDGFR‐α and almost most of the HA cases were negative for PDGFR‐β. Of the HSA cases, 55.6% were negative for PDGFR‐α and 63% were strongly positive for PDGFR‐β. Among the HA cases, 1 missense mutation was detected in PDGFR‐α exon 18 and 1 in PDGFR‐β exon 17. Two HSA cases had missense mutations in exon 14 and 1 in exon 17 of PDGFR‐β. Thus, genomic mutation of trans‐ or juxtamembrane regions of PDGFRs was not the main mechanism driving the activation of receptors in HSA and HA.     

Diurnal changes in plasma melatonin and the timing of reproductive onset in anestrous sheep fed melatonin.

Seasonally anestrous Suffolk ewes (n = 28) were randomly divided into 5 groups and treated with varying doses of melatonin as follows; Groups C (n = 4), M1 (n = 6), M2 (n = 6), M3 (n = 6), M4 (n = 6) of ewes were fed pellets containing 0, 1, 2, 3 and 4 mg melatonin, respectively, daily for 60 days from May 17 (Day 0). Following feeding of the pellets at 13.00 hr plasma levels of melatonin rapidly increased reaching the peak values within 30 min, which ranged from 92.0 to 292.7 pg/ml and were highly correlated with the dose of melatonin administered (r = 0.986, P less than 0.01). Maximum dose of melatonin (4 mg) produced an increase of plasma melatonin similar in magnitude to nocturnal peaks of endogenous secretion. The onset of ovulatory cyclicity, assessed from plasma progesterone profiles, was advanced by melatonin administration. The mean +/- SEM intervals from the commencement of melatonin treatment until the onset of ovulatory cyclicity were 53.0 +/- 5.8, 53.6 +/- 2.5, 42.0 +/- 5.6 and 44.3 +/- 4.3 days for the Groups M1, M2, M3 and M4, respectively, which were shorter (M1, P less than 0.05; M2, M3 & M4, P less than 0.01) than that for the Group C (72.5 +/- 1.4 days). The melatonin treatment also suppressed, in a dose related manner, the rise in plasma prolactin under the lengthening photoperiod. We conclude that the dose-related efficacy of melatonin could be ascribed to the difference in the diurnal profiles of circulating melatonin.     

Reference Values on Serum Biochemical Parameters of Brazilian Donkey (Equus asinus) Breed

Seventeen serum biochemical variables were determined in 40 donkeys of the Brazilian breed (34 females and 6 males) aged from 3 to 19 years. Mean ± standard deviation (SD) (minimum–maximum) values, obtained by automated analysis, were as follows: glucose, 58.35 ± 10.40 (44.00–90.00) mg/dL; cholesterol, 88.41 ± 9.86 (73.58–124.26) mg/dL; serum protein, 6.82 ± 0.40 (6.00–7.52) g/dL; albumin, 3.13 ± 0.21 (2.65–3.69) g/dL; creatinine, 1.80 ± 0.14 (1.51–2.19) mg/dL; urea, 24.25 ± 5.37 (14.12–34.39) mg/dL; lactate, 20.10 ± 4.58 (12.99–33.47) mg/dL; aspartate aminotransferase (AST), 295.81 ± 62.79 (173.71–466.07) IU/L; creatine kinase (CK), 158.00 ± 76.94 (51.69–440.33) IU/L; γ-glutamil-transferase (GGT), 45.82 ± 13.34 (26.17–86.38) IU/L; lactate dehydrogenase (LDH), 576.02 ± 156.32 (213.53–1162.81) IU/L; alkaline phosphatase (AP), 345.36 ± 65.90 (227.25–490.16) IU/L; calcium (Ca), 8.54 ± 0.18 (8.19–8.90) mg/dL; phosphorus (P), 2.76 ± 0.38 (1.99–3.97) mg/dL; chloride (Cl), 106.05 ± 3.20 (99.00–112.00) mEq/L; sodium (Na), 121.50 ± 4.14 (116.00–132.00) mEq/L; and potassium (K), 3.70 ± 0.42 (2.80–4.40) mEq/L. Comparisons of biochemical ranges obtained for the Brazilian donkey breed with reference ranges for other donkey breeds suggested that most values were similar. Biochemical values determined in the present study serve as reference ranges for donkey populations and can be used for health control and diagnosis of diseases.     

Stereotaxic approach to hypothalamic nuclei of the Shiba goat with radiographic monitoring

Practical method was devised for precise approach to hypothalamic nuclei in the Shiba goat. A stereotaxic instrument and a brain atlas with stereotaxic coordinates were developed. For an accurate placement of probes into specific hypothalamic regions a radiographic method was employed in which radio-opaque material was injected into the lateral ventricle and the ventricular outline was depicted. A sagittal diagram showing the arrangement of hypothalamic nuclei in relation to the brain ventricular system was constructed from the transverse stereotaxic atlas. This diagram was revealed extremely useful in pinpointing the target on the radiographs of lateral view. Precision of this method was evaluated in female Shiba goats (n = 4) by comparing radiographically estimated positions of hypothalamic nuclei with those histologically determined. Despite of cranial variability among individual animals these two parameters matched well each other in all the nuclei examined. Furthermore, chronic cannulae were implanted into different hypothalamic structures of one goat and the accuracy of their placement was confirmed histologically. Thus, it was revealed that the stereotaxy by aid of radiography herein described was accurate enough to apply to various neuroendocrinological studies in the Shiba goat.     

Effects of feeding high levels of cactus (Opuntia fícus-indica Mill) cladodes on urinary output and electrolyte excretion in goats

A study was conducted to determine the effects of feeding spineless cactus cladodes on diuresis and urinary electrolyte excretion in goats. Five bucks were used in a 5 × 5 Latin square experiment with 17-day periods. Experimental diets contained (g/kg dry matter (DM) basis) 370, 470, 570, 670, and 770 spineless cactus cladodes. Water consumption from feed and urine output increased linearly (P < 0.05) as the level of cactus cladodes in the diet increased. However, water intake from drinking was low and unaffected by cactus cladode level. Creatinine clearance and urinary Na excretion were similar for all dietary treatments while K excretion decrease linearly (P < 0.05) as the level of cactus cladodes in the diet increased. Feeding cactus cladodes caused diuresis and reduced urinary K excretion in goats. Possible reasons for these effects include water over-consumption from cactus cladodes and high dietary K intake.     

Ascorbic acid promotes graft-take in sweet pepper plants (Capsicum annuum L.)

The cause of the low survival rate of sweet pepper plants after grafting was investigated and compared with findings of eggplant and tomato plants, and the promotive effect of ascorbic acid (AA) was determined. Sweet pepper plants formed less callus at the cut surfaces of the stem compared to eggplant and tomato plants. Foliar application with 100 mg L⁻¹ AA promoted callus formation at the cut surfaces of the scion stems and improved the survival rate after grafting. We suggest that the low survival rate of grafted sweet pepper plants is caused by their low rate of callus formation, and that AA can be used to promote graft-take through the acceleration of callus formation at the cut surfaces of the stems.     

Leaf morphology and photosynthetic adjustments among deciduous broad-leaved trees within the vertical canopy profile

Photosynthetic acclimation of deciduous broad-leaved tree species was studied along a vertical gradient within the canopy of a multi-species deciduous forest in northern Japan. We investigated variations in (1) local light regime and CO2 concentration ([CO2]), and (2) morphological (area, thickness and area per mass), biochemical (nitrogen and chlorophyll concentrations) and physiological (light-saturated photosynthetic rate) attributes of leaves of seven major species on three occasions (June, August and October). We studied early successional species, alder (Alnus hirsuta (Spach) Rupr.) and birch (Betula platyphylla var. japonica (Miq.) Hara); gap phase species, walnut (Juglans ailanthifolia Carrière) and ash (Fraxinus mandshurica var. japonica Rupr.); mid-successional species, basswood (Tilia japonica (Miq.) Simonk.) and elm (Ulmus davidiana var. japonica (Rehd.) Nakai); and the late-successional species, maple (Acer mono Bunge). All but maple initiated leaf unfolding from the lower part of the crown. The [CO2] within the vertical profile ranged from 320-350 ppm in the upper canopy to 405-560 ppm near the ground. The lowest and highest ambient [CO2] occurred during the day and during the night, respectively. This trend was observed consistently during the summer, but not when trees were leafless. Chlorophyll concentration was positively related to maximum photosynthetic rate within, but not among, species. Leaf senescence started from the inner part of the crown in alder and birch, but started either in the outer or top portion of the canopy of ash, basswood and maple. Chlorophyll (Chl) to nitrogen ratio in leaves increased with decreasing photon flux density. However, Chl b concentration in all species remained stable until the beginning of leaf senescence. Maximum photosynthetic rates observed in sun leaves of early successional species, gap phase or mid-successional species, and late successional species were 12.5-14.8 micromol m(-2) s(-1), 4.1-7.8 micromol m(-2) s(-1) and 3.1 micromol m(-2) s(-1), respectively.     

Functional relationships between crown morphology and within-crown characteristics of understory saplings of three codominant conifers in a subalpine forest in central Japan

Light-related plasticity of crown morphology and within-crown characteristics were investigated in understory sun and shade saplings of three codominant subalpine conifers, Abies mariesii M.T. Mast., Abies veitchii Lindl. and Picea jezoensis var. hondoensis (Mayr) Rehd. Compared with those of sun saplings, current-year shoots of shade saplings allocated less biomass to needles, resulting in less dense needle packing and hence less mutual needle shading. The proportion of lateral branch biomass in foliage was either similar in sun and shade saplings or greater in shade saplings, depending on the species, suggesting that, over the lifetime of a branch, greater needle longevity in shade compensates for reduced biomass investment in needles of current-year shoots of shade saplings. Saplings with slower-growing branches tended to have greater needle life spans, suggesting that plasticity of branch growth rate and plasticity of needle life span are interdependent. Both Abies species showed greater light-related plasticity of needle life span and branch growth than P. jezoensis. The greater shade tolerance of the Abies species derives from their broad flattened crowns with slow-growing branches. This type of crown development incurs substantial support costs, but the long needle life span of shade saplings of the Abies species compensates, at least in part, for their low annual investment in foliage, especially in the case of A. mariesii, which has a longer needle life span and slower-growing and stouter branches than A. veitchii. Compared with the Abies species, P. jezoensis had a less plastic crown morphology, and less variability of needle life span and branch growth in response to light, resulting in lower shade tolerance. However, compared with the flattened crown of Abies shade saplings, the conical crown of P. jezoensis saplings imposes a smaller support cost, making this species better adapted to rapid height growth than to survival in shade.