Light-dependent quenching of chlorophyll fluorescence in pea chloroplasts induced by adenosine 5′-triphosphate

Addition of ATP to chloroplasts causes a reversible 25–30% decrease in chlorophyll fluorescence. This quenching is light-dependent, uncoupler insensitive but inhibited by DCMU and electron acceptors and has a half-time of 3 minutes. Electron donors to Photosystem I can not overcome the inhibitory effect of DCMU, suggesting that light activation depends on the reduced state of plastoquinone. Fluorescence emission spectra recorded at ?196°C indicate that ATP treatment increases the amount of excitation energy transferred to Photosystem I. Examination of fluorescence induction curves indicate that ATP treatment decreases both the initial (F_o) and variable (F_v) fluorescence such that the ratio of F_v to the maximum (F_m) yield is unchanged. The initial sigmoidal phase of induction is slowed down by ATP treatment and is quenched 3-fold more than the exponential slow phase, the rate of which is unchanged. A plot of F_v against area above the induction curve was identical plus or minus ATP. Thus ATP treatment can alter quantal distribution between Photosystems II and I without altering Photosystem II-Photosystem II interaction. The effect of ATP strongly resembles in its properties the phosphorylation of the light-harvesting complex by a light activated, ATP-dependent protein kinase found in chloroplast membranes and could be the basis of physiological mechanisms which contribute to slow fluorescence quenching in vivo and regulate excitation energy distribution between Photosystem I and II. It is suggested that the sensor for this regulation is the redox state of plastoquinone.     

ATP oscillation inPhysarum plasmodium

Summary Using bioluminescence of luciferin-luciferase, we showed that ATP leaked out rhythmically from a strand segment ofPhysarum plasmodium made permeable with caffeine-arsenate. With simultaneous measurement of isometric tension rhythm of the strand, it was revealed that the period and phase of oscillation in ATP leakage correspond well with those of tension production. Further, microinjection of luciferin-luciferase into the plasmodial strand indicated that the intracellular luminescence of luciferin-luciferase also oscillates with the same period and in the same phase as the tension rhythm.The free ATP concentration in a homogenate ofPhysarum plasmodium was of the order of 10 M, but if the homogenate was heated in boiling water, the intensity of luminescence suddenly increased 10–100 fold. ATP available for mechanical workin vivo is thus supposed to be at a much lower level than the total average, which was found in the range of 0.2–0.7 mM.     

Effects of feeding and starvation on growth and swimming activity in an obligatory air-breathing fish

Reared in (tubular) aquaria containing different depths of water, Ophiocephalus striatus (0.7 g, 4.5 cm body length), an obligatory air-breathing tropical fish, swam long or short distances to enable themselves to exchange atmospheric air. In each tested depth (2.5, 5.0, 15.5, 31.0 and 40.0 cm) series, one group was starved, while the other was fed ad libitum twice a day on fish muscle. In the shallowest water (2.5 cm depth), the feeding group surfaced 1,294 times, travelling 64.7 m at an energy cost of 20.4 mg dry fish substance/g live fish/day, against those exposed to the deepest water (40 cm depth), which expended 35.8 mg/g/day, swimming 1,503.4 m on 1,879 visits to the surface. The starving group surfaced only 482 times, travelling 24.1 m at an expense of 5.8 mg/g/day in the shallowest water, while those at 40 cm depth surfaced 504 times, swimming 403.2 m at an energy cost of 7.4 mg/g/day. Owing to the sustained swimming activity and the consequent fatigue, the test individuals belonging to both groups in all the tested series hang to the surface for a definite interval, repaying the O₂ debt. Observations were also made to assess the duration of hanging to precisely estimate the distance travelled. Irrespective of changes in depths of water, the duration of hanging to surface was only 3.0 hr/day for the feeding groups, while it was as much as 15.5 hr/day for the starving groups. The maximum sustained metabolic level of O.striatus reared in 40 cm depth was equivalent to 1.23 ml O₂/g/hr, which is about 2 times higher than the value reported for the active metabolism of swimming Oncorhynchus nerka at 15°C in Brett's (1964) respirometer. O.striatus reared in 2.5 cm depth fed 32.0 mg and converted 6.7 mg dry food/g live fish/day, while those exposed to the deepest water fed 49.1 mg, but converted only 5.5 mg/g/day. Culturing obligatory air-breathing fishes in shallow waters will be advantageous.     

An optimal dietary non-phytate phosphorus level of broilers fed a conventional corn–soybean meal diet from 4 to 6 weeks of age

It is imperative to evaluate precise nutrient requirements of animals in order to optimize productivity and minimize feed cost and nutrient excretions. The current non-phytate phosphorus (NPP) recommendation for broilers is based on the papers published 30 years ago. However, today’s commercial birds are quite different from those before 30 years. Therefore, the present experiment was conducted with growing male broiler chickens to evaluate an optimal dietary NPP level of broiler chickens fed a conventional corn–soybean meal diet from 4 to 6 weeks of age. The 1-day-old chicks were fed corn–soybean meal diet containing 0.39% NPP from 1 to 3 weeks of age. At 22 days of age, 360 birds were selected and randomly allotted by BW to one of 10 dietary treatments with six replicate cages of six birds per cage for each treatment. Birds were fed the P-unsupplemented corn–soybean meal basal diet and the basal diet supplemented with inorganic P as CaHPO₄·H₂O ranging from 0.00% to 0.45% with 0.05% increment from 4 to 6 weeks of age. The dietary NPP levels were 0.09%, 0.14%, 0.20%, 0.24%, 0.30%, 0.34%, 0.38%, 0.45%, 0.49% and 0.54%, respectively, and the dietary Ca level was fixed at 0.90% for all treatments. The results showed that average daily gain, serum inorganic P concentration, tibia bone strength, tibia ash percentage and P percentage, tibia bone mineral content (BMC) and density (BMD), middle toe ash percentage and P percentage, middle toe BMC, total body BMC and BMD were affected (P<0.0001) by dietary NPP level, and increased linearly (P<0.0001) and quadratically (P<0.003) as dietary NPP levels increased. Optimal dietary NPP levels estimated based on fitted broken-line models (P<0.0001) of the above indices are 0.21%, 0.29%, 0.29%, 0.29%, 0.29%, 0.31%, 0.29%, 0.30%, 0.27%, 0.29% and 0.28%, respectively. It is suggested that the total body BMC and BMD, and middle toe ash P and BMC might be new, sensitive and non-invasive criteria to evaluate the dietary NPP requirements of broilers. The optimal dietary NPP level would be 0.31% for broiler chickens fed a conventional corn–soybean meal diet from 4 to 6 weeks of age.     

Combined effect of glycerol concentration and cooling velocity on motility and acrosomal integrity of boar spermatozoa frozen in 0.5 ml straws

The interaction of glycerol concentrations of 0-10% and cooling rates from 1 to 1,500 degrees C/min with boar spermatozoa motility and acrosomal integrity (proportion of spermatozoa with normal apical ridge) was studied after thawing 0.5 ml straws at a constant rate. While increasing the glycerol concentration from 0 to 4% progressively improved motility, the percentage of spermatozoa with a normal apical ridge gradually decreased. The magnitudes of the respective changes depended on cooling rate. A peak value of 48.1% and rating 3.8 were obtained in semen protected with 4% glycerol, frozen at 30 degrees C/min. Increasing the glycerol levels above 6% resulted in a gradual decrease in motility. The proportion of spermatozoa with normal apical ridge was highest in semen protected with 0-1% glycerol after cooling at 30 degrees C/min (64.4% and 66.1%, respectively), but at these glycerol concentrations the percentage of motile spermatozoa was low. At the 30 degrees C/min cooling rate, the decline in the proportion of cells with normal apical ridge due to increasing the glycerol levels to 3 and 4% was relatively slow (57.3% and 49.4%, respectively). Cooling at 1 degrees C/min was detrimental to acrosomal integrity, which decreased with increasing glycerol concentration, in contrast to increasing motility, which even at its maximum, remained low. The direct plunging of straws into liquid nitrogen (1,500 degrees C/min) resulted in damaged acrosomes in all spermatozoa with the total loss of motility. Balancing motility and acrosomal integrity, freezing boar semen protected with 3% glycerol by cooling at 30 degrees C/min resulted in optimal survival for boar semen frozen in 0.5 ml French straws.     

SHCBP1:乳腺癌治疗候选靶点和潜在预后标志物

探讨SHCBP1在乳腺癌及其不同亚型中的表达特征和预后价值。利用Oncomine, bc-GenExMiner v4.2, Kaplan-Meier plotter, GSE41994及STRING数据库分析乳腺癌病人SHCBP1的mRNA表达水平及其与乳腺癌患者生存率的相关性等。结果:(1)乳腺癌组织中SHCBP1的mRNA水平明显高于正常组织样本,但不同分子亚型的乳腺癌患者中SHCBP1的表达水平不同;(2)SHCBP1表达升高可导致Luminal A亚型更短的无转移生存期(Distant metastasis-free survival,DMFS)和无病生存期(Disease-free survival, DFS);(3)在雌激素受体阳性(ER+)、孕酮受体阳性(PR+)亚型、仅接受辅助化疗、仅接受辅助化疗的ER(+)与接受包括或排除内分泌治疗乳腺癌患者中,SHCBP1表达升高则无复发生存率(Relapse-free survival,RFS)显著缩短;(4)SHCBP1和CDC45 mRNA表达水平之间呈正相关,并可能与KIF23等10种蛋白相互作用。结论:SHCBP1是一种很有前景的乳腺癌预后指标和潜在的治疗靶点。