Effects of understory removal on soil respiration and microbial community composition structure in a Chinese fir plantation

Aims Soil respiration (R_s) is the largest fraction of carbon flux in forest ecosystems, but the effects of forest understory removal on R_s in Chinese fir (Cunninghamia lanceolate) plantations is poorly understood. In order to quantify the effects of forest understory removal on R_s and microbial community composition, a field experiment was conducted in a subtropical Chinese fir plantation. Methods Forest understory was removed manually in June 2012. R_s was measured monthly using a LI-COR 8100 infrared gas analyzer from July 2012 through July 2014. Soil temperature and moisture were also measured at 5 cm depth at the time of R_s measurements. Surface soil (0-10 cm) samples were collected in July 2013 and 2014, respectively, and the soil microbial community structures were determined by phospholipid fatty acids (PLFAs) analysis. Important findings R_s decreased by 32.8% over a two-year period following understory removal (UR), with a greater rate of decrease in the first year (42.9%) than in the second year (22.2%). The temperature sensitivity of R_s was affected by UR, and was 2.10 and 1.87 in the control and UR plots, respectively. UR significantly reduced the concentration of fungal PLFAs by 18.3%, but did not affect the concentration of bacterial PLFAs, resulting in an increase in the fungal:bacterial ratio; it significantly increased the concentration of gram-positive bacterial PLFAs by 24.5%, and the ratio of gram-positive to gram-negative bacterial PLFAs after one year of treatment, but decreased the concentration of gram-positive bacterial PLFAs by 9.4% and the ratio of gram-positive to gram-negative bacterial PLFAs after two years of treatment. The results suggested that R_s and microbial community composition were both affected by UR in Chinese fir plantation, and the effects were dependent of the duration following the UR treatment.     

林下植被剔除对杉木林土壤呼吸和微生物群落结构的影响

以湖南会同地区26年生杉木(Cunninghamia lanceolata)人工林为研究对象, 探讨剔除林下植被对土壤呼吸和微生物群落结构的影响。2012年6月将林下植被剔除后, 2012年7月-2014年7月每月测定一次土壤呼吸速率、5 cm土壤温度和含水量, 并分别于2013年7月和2014年7月测定了土壤微生物群落结构和土壤养分数据。研究结果表明: 杉木人工林土壤呼吸具有明显的季节变化规律, 且与5 cm深处的土壤温度呈极显著的正相关关系。林下植被剔除两年内土壤呼吸平均下降了32.8%, 2012年7月-2013年6月下降了42.9%, 2013年7月-2014年7月下降了22.2%。根据土壤呼吸与温度拟合的指数方程所计算出的土壤呼吸的温度敏感性Q₁₀值在对照区为2.10, 林下植被剔除区为1.87, 说明在杉木人工林系统中林下植被剔除2年降低了土壤呼吸的温度敏感性。此外, 林下植被剔除也改变了土壤微生物群落结构。林下植被剔除1年后, 土壤细菌的浓度没有发生改变, 但真菌的浓度降低, 导致真菌与细菌的浓度比值下降。此外, 革兰氏阳性细菌(G⁺)的浓度及其与革兰氏阴性菌(G^-)的比值升高。林下植被剔除2年后, G⁺浓度和G⁺与G^-的浓度比值降低。该研究表明林下植被剔除可以降低土壤呼吸, 从而减少土壤向大气中释放碳; 同时可改变土壤微生物群落结构, 而且其效应受作用时间的影响。     

In situ inactivation of E. coli uridylylation cycle is independent of the state of covalent modification of the components of the glutamine synthetase cascade

Permeabilization of nitrogen-starved cells of Escherichia coli W with Lubrol WX leads to a selective inactivation of the uridylyl-removing uridylyltransferase (UR/ UTase) enzyme of the glutamine synthetase (GS) cascade system; whereas similar treatment does not affect activity of UR/UTase in cells grown under conditions of nitrogen excess (10 mm glutamine) (Mura, U., and Stadtman, E. R. (1981) J. Biol. Chem.256, 13014–13021). The possibility that susceptibility to Lubrol inactivation is related to differences in the state of adenylylation of GS and/or in the state of uridylylation of the P_II protein was investigated. Permeabilized cells from nitrogen sufficient as well as from nitrogen-limited growth medium were exposed to Lubrol after prior incubation under conditions that lead to high or low states of GS adenylylation and high or low P_IID/P_IIA ratios. Integrity of UR/UTase was monitored by measuring the capacity of UTP to stimulate the deadenylylation of GS in situ. The results showed that the inactivation of UR/UTase by Lubrol is not affected by the states of GS adenylylation or P_II uridylylation.     

Acute and chronic effects of exercise on tissue sensitivity to glucocorticoids.

The aim of this study was to address the effect of endurance training on tissue sensitivity to glucocorticoids (GC) in both resting and exercising conditions. In vitro dexamethasone inhibition of LPS-induced interleukin-6 secretion in cultures of peripheral monocytes was compared in untrained subjects (UT) and in endurance-trained men (ET) at the end of a 2-h run and during exercise recovery. We demonstrated an in vitro plasticity of sensitivity of monocytes to GC in ET men, superimposed to changes in systemic cortisol concentrations (plasma and saliva). Compared with sedentary men, similar resting cortisol levels in ET men are associated with decreased sensitivity of monocytes to GC 8 and 24 h after the end of the last training session (P < 0.05, ET vs. UT). Moreover, in these ET subjects, an acute bout of exercise increased the sensitivity of monocytes to GC (at 1000 and 1200; ET vs. UT, P > 0.05). This acute exercise-induced increase in tissue sensitivity to GC, which is synchronous with activation of the hypothalamo-pituitary adrenal axis, may act to shut off muscle inflammatory reaction and cytokine synthesis and then decrease exercise-induced muscle damage or inflammatory response. By contrast, the decreased sensitivity of monocytes to GC reported in ET men 24 h after the last bout of exercise may be related to the process of desensitization that may act to protect the body from prolonged, exercise-induced cortisol secretion. These acute and chronic effects of exercise on tissue sensitivity to GC demonstrate an adaptation of the hypothalamo-pituitary adrenal axis to repeated and prolonged exercise-induced increases in GC secretion.     

Coevolution of the hepatitis C virus polyprotein sites in patients on combined pegylated interferon and ribavirin therapy

Genotype-specific sensitivity of the hepatitis C virus (HCV) to interferon-ribavirin (IFN-RBV) combination therapy and reduced HCV response to IFN-RBV as infection progresses from acute to chronic infection suggest that HCV genetic factors and intrahost HCV evolution play important roles in therapy outcomes. HCV polyprotein sequences (n = 40) from 10 patients with unsustainable response (UR) (breakthrough and relapse) and 10 patients with no response (NR) following therapy were identified through the Virahep-C study. Bayesian networks (BNs) were constructed to relate interrelationships among HCV polymorphic sites to UR/NR outcomes. All models showed an extensive interdependence of HCV sites and strong connections (P ≤ 0.003) to therapy response. Although all HCV proteins contributed to the networks, the topological properties of sites differed among proteins. E2 and NS5A together contributed ～40% of all sites and ～62% of all links to the polyprotein BN. The NS5A BN and E2 BN predicted UR/NR outcomes with 85% and 97.5% accuracy, respectively, in 10-fold cross-validation experiments. The NS5A model constructed using physicochemical properties of only five sites was shown to predict the UR/NR outcomes with 83.3% accuracy for 6 UR and 12 NR cases of the HALT-C study. Thus, HCV adaptation to IFN-RBV is a complex trait encoded in the interrelationships among many sites along the entire HCV polyprotein. E2 and NS5A generate broad epistatic connectivity across the HCV polyprotein and essentially shape intrahost HCV evolution toward the IFN-RBV resistance. Both proteins can be used to accurately predict the outcomes of IFN-RBV therapy.     

Anticonvulsive and free radical scavenging actions of two herbs, Uncaria rhynchophylla (MIQ) Jack and Gastrodia elata Bl., in kainic acid-treated rats

Uncaria rhynchophylla (Miq.) Jack (UR) and Gastrodia elata BI. (GE) are traditional Chinese herbs that are usually used in combination to treat convulsive disorders, such as epilepsy, in China. The aim of this study was to compare the anticonvulsive and free radical scavenging activities of UR alone and UR in combination with GE in rats. For the in vitro studies, brain tissues from 6 male Sprague-Dawley (SD) rats were treated with 120 microg/ml kainic acid (KA), with or without varied concentrations of UR or UR plus GE. For the in vivo studies, male SD rats (6 per group) received intraperitoneal (i.p.) injection of KA 12 mg/kg to induce epileptic seizures and generation of free radicals, with or without oral administration of UR 1 g/kg alone or UR 1 g/kg plus GE 1 g/kg. Epileptic seizures were verified by behavioral observations, and electroencephalography (EEG) and electromyography (EMG) recordings. These results showed that UR alone decreased KA-induced lipid peroxide levels in vitro, whereas UR plus GE did not produce a greater effect than UR alone. UR significantly reduced counts of wet dog shakes (WDS), paw tremor (PT) and facial myoclonia (FM) in KA-treated rats and significantly delayed the onset time of WDS, from 27 min in the control group to 40 min in the UR group. UR plus GE did not inhibit seizures more effectively than UR alone, but did further prolong the onset time of WDS to 63 min (P < 0.05 vs. UR alone). UR alone reduced the levels of free radicals in vivo, as measured by lipid peroxidation in the brain and luminol-chemiluminescence (CL) counts and lucigenin-CL counts in the peripheral whole blood, but the combination of GE and UR did not reduce free radical levels more markedly than UR alone. In conclusion, our results indicate that UR has anticonvulsive and free radical scavenging activities, and UR combined with GE exhibit greater inhibition on the onset time of WDS than UR alone. These findings suggest that the anticonvulsive effects of UR and GE may be synergistic. However, the mechanism of interaction between UR and GE remains unknown.     

Nucleotide sequence of avian sarcoma virus UR2 and comparison of its transforming gene with other members of the tyrosine protein kinase oncogene family.

The genome of avian sarcoma virus UR2 was completely sequenced and found to have a size of 3,165 nucleotides. The UR2-specific transforming sequence, ros, with a length of 1,273 nucleotides, is inserted between the truncated gag gene coding for p19 and the env gene coding for gp37 of the UR2AV helper virus. The deduced amino acid sequence for the UR2 transforming protein P68 gives a molecular weight of 61,113 and shows that it is closely related to the oncogene family coding for tyrosine protein kinases. P68 contains two distinctive hydrophobic regions that are absent in other tyrosine kinases, and it has unique amino acid changes and insertions within the conserved domain of the kinases. These characteristics may modulate the activity and target specificity of P68.     

Antioxidant activity and protective effects of Uncaria rhynchophylla extracts on t-BHP-induced oxidative stress in Chang cells

Uncaria rhynchophylla (UR) is widely used as a traditional Chinese herbal medicine. However, there are few studies on antioxidant activity of UR extracts. The study was aimed at determining the antioxidant activity, the contents of polyphenol and flavonoid of UR extracts. Various assays were employed to evaluate the antioxidant property of water and ethanol extracts from the UR, compared to those of the other natural and synthetic antioxidants. UR extracts had high total phenolic contents in both the water (160 ± 2.32 mg GAE/g extracts) and ethanol extracts (190.2 ± 3.16 mg GAE/g extracts). In addition, total flavonoid contents were high in both the water extracts (154 ± 1.47 mg CE/g extracts) and ethanol extracts (184.2 ± 2.41 CE/g extracts). Specially, DPPH radical scavenging activity of ethanol extracts from UR were similar with vitamin C as a positive control. In addition, antioxidant capacity in ferric reducing antioxidant power (FRAP) were higher than that for BHT, which was used as a positive control. The antioxidant activity of extracts from UR showed stronger activity than those of vitamin C and α-tocopherol in ferric thiocyanate (FTC) and thiobarbituric acid (TBA) methods. The ethanol extracts of UR protected on H₂O₂-induced DNA damage. In addition, cytoprotective and anti-apoptotic effect of ethanol extracts from UR was also investigated in t-BHP-induced hepatotoxicity. Therefore, these results indicate that UR extracts have antioxidant properties through its ability to enhance the cell viability, mediation of production of ROS. The UR extracts could be suitable as an antioxidant in the food industry.     

Genetic structure, transforming sequence, and gene product of avian sarcoma virus UR1

We analyzed the genetic structure and gene products of the newly isolated avian sarcoma virus UR1, which recently has been shown to be replication defective and to contain no sequences homologous to the src gene of Rous sarcoma virus. The sizes of the genomic RNAs of UR1 and its associated helper virus, UR1AV, were determined to be 29S and 35S (5.9 and 8.5 kilobases), respectively, by gel electrophoresis and sucrose gradient sedimentation. RNase T1 oligonucleotide mapping of purified viral RNAs indicated that UR1 RNA contains eight unique oligonucleotides in the middle of the genome and shares four 5'-terminal and three 3'-terminal oligonucleotides with UR1AV RNA. The unique sequences of UR1 and Fujinami sarcoma virus were found to be closely related to each other by molecular hybridization of UR1 RNA with DNA complementary to the unique sequence of Fujinami sarcoma virus RNA, but minor differences were found by oligonucleotides fingerprinting. In the regions flanking the unique sequences, UR1 and Fujinami sarcoma viral RNAs contain distinct oligonucleotides, which are shared with oligonucleotides of the respective helper viral RNAs. Cell transformed with UR1 produce a single 29S RNA species which contains a UR1 unique sequence; this species is most likely the mRNA coding for the transforming protein. In UR1-transformed cells, a phosphoprotein fo 150,000 daltons (p150) was detected by immunoprecipitation with antiserum against gag proteins. p150 was associated with a protein kinase activity that was capable of phosphorylating p150 itself, immunoglobulin G of antiserum, and a soluble substrate, alpha-casein. This enzyme transferred phosphate exclusively to tyrosine residues of substrates in vitro, but p 150 labeled in vivo with 32P contained both phosphoserine and phosphotyrosine. The in vitro kinase reaction was not affected by the presence of cyclic AMP or cyclic GMP and strongly preferred Mn2+ over Mg2+. Thus, the properties of UR1 protein are almost identical to those of Fujinami sarcoma virus protein.     

Factors Affecting De Novo Urinary Retention after Holmium Laser Enucleation of the Prostate

Objective

Patients can experience urinary retention (UR) after Holmium laser enucleation of the prostate (HoLEP) that requires bladder distension during the procedure. The aim of this retrospective study is to identify factors affecting the UR after HoLEP.

Materials and Methods

336 patients, which underwent HoLEP for a symptomatic benign prostatic hyperplasia between July 2008 and March 2012, were included in this study. Urethral catheters were routinely removed one or two days after surgery. UR was defined as the need for an indwelling catheter placement following a failure to void after catheter removal. Demographic and clinical parameters were compared between the UR (n = 37) and the non-urinary retention (non-UR; n = 299) groups.

Results

The mean age of patients was 68.3 (±6.5) years and the mean operative time was 75.3 (±37.4) min. Thirty seven patients (11.0%) experienced a postoperative UR. UR patients voided catheter free an average of 1.9 (±1.7) days after UR. With regard to the causes of UR, 24 (7.1%) and 13 (3.9%) patients experienced a blood clot-related UR and a non-clot related UR respectively. Using multivariate analysis (p<0.05), we found significant differences between the UR and the non-UR groups with regard to a morcellation efficiency (OR 0.701, 95% CI 0.498–0.988) and a bleeding-related complication, such as, a reoperation for bleeding (OR 0.039, 95% CI 0.004–0.383) or a transfusion (OR 0.144, 95% CI 0.027–0.877). Age, history of diabetes, prostate volume, pre-operative post-void residual, bladder contractility index, learning curve, and operative time were not significantly associated with the UR (p>0.05).

Conclusions

De novo UR after HoLEP was found to be self-limited and it was not related to learning curve, patient age, diabetes, or operative time. Efficient morcellation and careful control of bleeding, which reduces clot formation, decrease the risk of UR after HoLEP.     

c‐FLIP is involved in erythropoietin‐mediated protection of erythroid‐differentiated cells from TNF‐α‐induced apoptosis

The TNF‐α (tumour necrosis factor) affects a wide range of biological activities, such as cell proliferation and apoptosis. Cell life or death responses to this cytokine might depend on cell conditions. This study focused on the modulation of factors that would affect the sensitivity of erythroid‐differentiated cells to TNF‐α. Hemin‐differentiated K562 cells showed higher sensitivity to TNF‐induced apoptosis than undifferentiated cells. At the same time, hemin‐induced erythroid differentiation reduced c‐FLIP (cellular FLICE‐inhibitory protein) expression. However, this negative effect was prevented by prior treatment with Epo (erythropoietin), which allowed the cell line to maintain c‐FLIP levels. On the other hand, erythroid‐differentiated UT‐7 cells – dependent on Epo for survival – showed resistance to TNF‐α pro‐apoptotic action. Only after the inhibition of PI3K (phosphatidylinositol‐3 kinase)‐mediated pathways, which was accompanied by negative c‐FLIP modulation and increased erythroid differentiation, were UT‐7 cells sensitive to TNF‐α‐triggered apoptosis. In summary, erythroid differentiation might deregulate the balance between growth promotion and death signals induced by TNF‐α, depending on cell type and environmental conditions. The role of c‐FLIP seemed to be critical in the protection of erythroid‐differentiated cells from apoptosis or in the determination of their sensitivity to TNF‐mediated programmed cell death. Epo, which for the first time was found to be involved in the prevention of c‐FLIP down‐regulation, proved to have an anti‐apoptotic effect against the pro‐inflammatory factor. The identification of signals related to cell life/death switching would have significant implications in the control of proliferative diseases and would contribute to the understanding of mechanisms underlying the anaemia associated with inflammatory processes.     

Molecular cloning and characterization of avian sarcoma virus UR2 and comparison of its transforming sequence with those of other avian sarcoma viruses.

Avian sarcoma virus UR2 and its associated helper virus, UR2AV , were molecularly cloned into lambda gtWES X lambda B by using unintegrated viral DNAs. One UR2 and several UR2AV clones were obtained. The UR2 DNA was subsequently cloned into pBR322. Both UR2 and UR2AV DNAs were tested for their biological activity by transfection onto chicken embryo fibroblasts. When cotransfected with UR2AV DNA, UR2 DNA was able to induce transformation of chicken embryo fibroblasts with a morphology similar to that of parental UR2 . UR2 -specific protein with kinase activity and UR2 -specific RNA were detected in the transfected cells. Transforming virus, UR2 ( UR2AV ), was produced from the doubly transfected cells. Five of the six UR2AV clones tested were also shown to be biologically active. The insert of the UR2 DNA clone is 3.4 kilobases in length and contains two copies of the long terminal repeat. Detailed restriction mapping showed that UR2 DNA shared with UR2AV DNA 0.8 kilobases of 5' sequence, including a portion of 5' gag, and 1.4 kilobases of 3' sequence, including a portion of 3' env. The UR2 transforming sequence, ros, is ca. 1.2 kilobases. No significant homology was found between v-ros and the conserved regions of v-src, v-yes, or v- abl . By contrast, a significant homology was found between v-ros and v-fps. The v-fps-related sequence was mapped within a 300-base-pair sequence in the middle of ros.     

The UII/UT System Mediates Upregulation of Proinflammatory Cytokines through p38 MAPK and NF-κB Pathways in LPS-Stimulated Kupffer Cells

The urotensin II (UII)/UII receptor (UT) system is closely related to immune inflammation. In acute liver failure (ALF), the UII/UT system can promote the production and release of proinflammatory cytokines, inducing an inflammatory injury response in liver tissue. However, the mechanism by which the hepatic UII/UT system promotes proinflammatory cytokine production and release is not clear. To solve this problem, we used primary Kupffer cells (KCs) as the model system in the current study. The results showed that after lipopolysaccharide (LPS) stimulation, KCs showed significantly increased expression and release of UII/UT and proinflammatory cytokines tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β). Pretreatment with urantide, which is a UT receptor antagonist, significantly inhibited the LPS-stimulated expression and release of UII/UT, TNF-α, and IL-1β by KCs. In addition, LPS stimulation induced nuclear p38 mitogen-activated protein kinase (MAPK) protein phosphorylation and expression of the nuclear nuclear factor κB (NF-κB) p65 subunit in KCs and enhanced the binding activity of NF-κB to DNA molecules, whereas urantide pretreatment significantly inhibited the LPS-stimulated nuclear expression and activity of these molecules in KCs. Therefore, our conclusion is that the UII/UT system mediates LPS-stimulated production and release of proinflammatory cytokine by KCs, and this mediating effect at least partially relies on the inflammatory signaling pathway molecules p38 MAPK and NF-κB.     

Cloning and characterization of an envelope-specific probe from xenotropic murine leukemia proviral DNA.

UR2 is a newly characterized avian sarcoma virus whose genome contains a unique sequence that is not related to the sequences of other avian sarcoma virus transforming genes thus far identified. This unique sequence, termed ros, is fused to part of the viral gag gene. The product of the fused gag-ros gene of UR2 is a protein of 68,000 daltons (P68) immunoprecipitable by antiserum against viral gag proteins. In vitro translation of viral RNA and in vivo pulse-chase experiments showed that P68 is not synthesized as a large precursor and that it is the only protein product encoded in the UR2 genome, suggesting that it is involved in cell transformation by UR2. In vivo, P68 was phosphorylated at both serine and tyrosine residues. Immunoprecipitates of P68 with anti-gag antisera had a cyclic nucleotide-independent protein kinase activity that phosphorylated P68, rabbit immunoglobulin G in the immune complex, and alpha-casein. The phosphorylation by P68 was specific to tyrosine of the substrate proteins. P68 was phosphorylated in vitro at only one tyrosine site, and the tryptic phosphopeptide of in vitro-labeled P68 was different from those of Fujinami sarcoma virus P140 and avian sarcoma virus Y73-P90. A comparison of the protein kinases encoded by UR2, Rous sarcoma virus, Fujinami sarcoma virus, and avian sarcoma virus Y73 revealed that UR2-P68 protein kinase is distinct from the protein kinases encoded by those viruses by several criteria. Our results suggest that several different protein kinases encoded by viral transforming genes have the same functional specificity and cause essentially the same cellular alterations.     

模拟氮沉降和灌草去除对杉木人工林地土壤微生物群落结构的影响

土壤微生物是陆地生态系统重要的分解者和地上-地下相互作用的纽带。本文以亚热带杉木(Cunninghamia lanceolateata)人工林为对象, 通过模拟林冠层氮沉降和林下灌草去除, 设置4种处理, 包括: 对照(CK)、灌草去除(UR)、氮沉降(N)和氮沉降加灌草去除(N × UR)的野外控制实验, 研究土壤微生物群落结构的响应。本实验分别于2016年4月(春季)和10月(秋季)采集0-10 cm层土壤样品, 运用磷脂脂肪酸法(PLFAs)分析土壤微生物群落结构。结果表明: (1) 10月份土壤微生物总PLFAs量及其他类群土壤微生物PLFAs量显著高于4月份(P < 0.05), 真菌/细菌比值没有显著差异。土壤微生物PLFAs中细菌占优势, 其次为真菌, 放线菌的占比最小; (2)相比CK处理, UR处理下土壤微生物总PLFAs量、细菌PLFAs量、革兰氏阴性菌PLFAs量和放线菌PLFAs量有增加趋势, 但未达到显著差异水平(P > 0.05); (3)相对CK, UR、N和N × UR处理降低了4月份土壤微生物多样性(H°)和均匀度指数(J), 但提高了10月份土壤微生物多样性指数; (4)冗余分析表明, 土壤硝态氮和总磷含量与土壤微生物群落之间呈现显著相关。本研究表明土壤微生物PLFAs在各处理下都表现出明显的季节动态; 短期内林下灌草去除对土壤微生物PLFAs影响表现出一定的促进作用, 氮沉降对土壤微生物群落影响还不甚明显, 需要长期的监测研究来评估两者及其交互作用对土壤微生物群落及其功能的影响。     

Changes in the synthesis and phosphorylation of cellular proteins in chick fibroblasts transformed by two avian sarcoma viruses

35S- and 32P-labeled proteins from control chick embryo fibroblasts and from fibroblasts transformed by UR2 sarcoma virus, or by a temperature-sensitive mutant (tsLA29) of Rous sarcoma virus, were separated by two-dimensional electrophoresis on giant gels to detect transformation-specific changes in protein synthesis and total phosphorylation. A nontransforming avian retrovirus, UR2-associated virus (UR2AV), was also studied. Virus-coded proteins appear in whole cell lysates of all infected cells. The structural proteins can be identified by comparison with proteins immunoprecipitated with antivirus serum. The transforming proteins pp60src and p68ros, present in cells transformed with Rous sarcoma virus and UR2, respectively, are phosphorylated in vivo. Eighteen increases and eight decreases in cellular phosphoproteins are associated with transformation, and revert toward normal levels when cells infected with tsLA29 are incubated at 42 degrees C. These changes are more extensive than previously reported, but none represent new phosphorylations, since all phosphoproteins seen in transformed cells also appear to be phosphorylated to a certain extent in control cells. Fifteen cellular proteins show increased relative rates of synthesis apparently related either to transformation or to growth at 42 degrees C. Four other proteins are increased exclusively in cells incubated at 42 degrees C, but not at 37 degrees C, whether transformed or not. Eleven additional increases in the synthesis of cellular proteins, many quite large, and one seemingly a de novo induction, appear to be specific for transformation. These changes occur in cells transformed by either UR2 or Rous sarcoma virus at 37 degrees C, do not occur with UR2AV infection, and tend to revert in cells infected with tsLA29 incubated at 42 degrees C. These 11 changes may represent increases in cellular gene expression that are related specifically to the maintenance of the transformed state.     

The phenotypic spectrum of GLI3 morphopathies includes autosomal dominant preaxial polydactyly type-IV and postaxial polydactyly type-A/B; No phenotype prediction from the position of GLI3 mutations.

Functional characterization of a gene often requires the discovery of the full spectrum of its associated phenotypes. Mutations in the human GLI3 gene have been identified in Greig cepalopolysyndactyly, Pallister-Hall syndrome (PHS), and postaxial polydactyly type-A (PAP-A). We studied the involvement of GLI3 in additional phenotypes of digital abnormalities in one family (UR003) with preaxial polydactyly type-IV (PPD-IV), three families (UR014, UR015, and UR016) with dominant PAP-A/B (with PPD-A and -B in the same family), and one family with PHS. Linkage analysis showed no recombination with GLI3-linked polymorphisms. Family UR003 had a 1-nt frameshift insertion, resulting in a truncated protein of 1,245 amino acids. A frameshift mutation due to a 1-nt deletion was found in family UR014, resulting in a truncated protein of 1,280 amino acids. Family UR015 had a nonsense mutation, R643X, and family UR016 had a missense mutation, G727R, in a highly conserved amino acid of domain 3. The patient with PHS had a nonsense mutation, E1147X. These results add two phenotypes to the phenotypic spectrum caused by GLI3 mutations: the combined PAP-A/B and PPD-IV. These mutations do not support the suggested association between the mutations in GLI3 and the resulting phenotypes. We propose that all phenotypes associated with GLI3 mutations be called "GLI3 morphopathies," since the phenotypic borders of the resulting syndromes are not well defined and there is no apparent genotype-phenotype correlation.     

Influence of training and maturity status on the cardiopulmonary responses to ramp incremental cycle and upper body exercise in girls

It has been suggested that the potential for training to alter the physiological responses to exercise in children is related to a "maturational threshold". To address this, we investigated the interaction of swim-training status and maturity on cardiovascular and metabolic responses to lower and upper body exercise. Twenty-one prepubertal [Pre: 11 trained (T), 10 untrained (UT)], 30 pubertal (Pub: 14 T, 16 UT), and 18 postpubertal (Post: 8 T, 10 UT) girls completed ramp incremental exercise on a cycle and an upper body ergometer. In addition to pulmonary gas exchange measurements, stroke volume and cardiac output were estimated by thoracic bioelectrical impedance, and muscle oxygenation status was assessed using near-infrared spectroscopy. All T girls had a higher peak O(2) uptake during cycle (Pre: T 49 ± 5 vs. UT 40 ± 4; Pub: T 46 ± 5 vs. UT 36 ± 4; Post: T 48 ± 5 vs. UT 39 ± 8 ml·kg(-1)·min(-1); all P < 0.05) and upper body exercise (Pre: T 37 ± 6 vs. UT 32 ± 5; Pub: T 36 ± 5 vs. UT 28 ± 5; Post: T 39 ± 3 vs. UT 28 ± 7 ml·kg(-1)·min(-1); all P < 0.05). T girls also had a higher peak cardiac output during both modalities, and this reached significance in Pub (cycle: T 21 ± 3 vs. UT 18 ± 3; upper body: T 20 ± 4 vs. UT 15 ± 4 l/min; all P < 0.05) and Post girls (cycle: T 21 ± 4 vs. UT 17 ± 2; upper body: T 22 ± 3 vs. UT 18 ± 2 l/min; all P < 0.05). None of the measured pulmonary, cardiovascular, or metabolic parameters interacted with maturity, and the magnitude of the difference between T and UT girls was similar, irrespective of maturity stage. These results challenge the notion that differences in training status in young people are only evident once a maturational threshold has been exceeded.