[Evaluation of primary tinnitus with fine puretone test and estimate of the technique].

Objective:To evaluate the relative factors of the severity and characteristics of primary tinnitus patients with fine puretone test(FPT) by three kinds of octaves, and to estimate the clinical application of this technique.Method:All subjects(n=90, ears=108), diagnosed with primary tinnitus, were recruited during July 2017 to February 2018 from the out-patients of Nanfang Hospital. In the prospective design, the patients were randomly divided into group 1/3 octave(n=30, ears=35), group 1/12 octave(n=30, ears=37) and group 1/24 octave(n=30, ears=36). Then tinnitus handicap inventory(THI), FPT, time and degree of residual inhibition(RI), time and degree of hyposensitization test, time attenuation test of masking(TATM) were used for evaluating the diseased and healthy ear performed separately.Result:Eighty-six patients were taken THI. The severity of tinnitus was no significant correlation with the gender,ear,sleep time and working time(P>0.05),but positive correlation with age(r=0.274, P=0.011) and duration of tinnitus(r=0.239, P=0.026). The selfassessment loudness score(SALS)(r=0.542,P=0.000) and the visual analogue score(VAS)(r=0.529, P=0.000) showed a moderate positive correlation. In the 1/24 octave, except for the TATM of the diseased ear group, the results of other groups were statistically different from the 1/3 octave(P<0.05), and much suitable than the 1/3 octave. Moreover, the results showed that the time of RI in the left and diseased ear group, the time of HT in the right, left and diseased ear group, the degree of HT in the right ear group were statistically different from the 1/12 octave(P<0.05) respectively, and much suitable than the 1/12 octave. The 1/12 octave was only statistically different from the 1/3 octave in TATM of the diseased ear group(P<0.05), and much suitable than the 1/3 octave.Conclusion:This study shows that gender, ear, sleep time, and working time had no significant effect on the severity of tinnitus, the age and duration of tinnitus have certain effects on the severity of tinnitus, the SALS, VAS score and the severity of tinnitus are moderately positively correlated. The 1/24 octave is significantly much suitable than the 1/3 octave and the 1/12 octave. And the technique of FPT is beneficial to improve the reliability and accuracy of the tinnitus evaluation.

[Application of technique of HRCT and multipliate plane reconstruction in analysis of image features of superior semicircular canal dehiscence].

Objective: Application of technique of high resolution CT (HRCT) and multipliate plane reconstruction, analyzed the image features of suspicious superior semicircular canal dehiscence (SSCD) to improve the understanding of the SSCD.Method:From January 2016 to April 2017, a total of 230 adult patients who checked temporal bone HRCT were collected in this retrospective study, of which 160 cases (320 ears) of the non-SSCD, 73 cases were male (146 ears), 87 cases were female (174 ears), aged 18 to 70 years old; 70 cases (113 ears) were suspicious diagnosed with SSCD, 33 cases were male (55 ears), 37 cases were female (58 ears), and 18 to 71 years old. The thin section CT scan of the temporal bone translocation was performed on all subjects, then the CT post processing workstation was used for multipliate plane reconstruction (MPR), and the main rows of coronal and oblique sagittal image reconstruction was performed. To observe, measure and record the HRCT features and datas of the suspicious SSCD and non-SSCD, then analyze the collected data.Result:There were 160 cases (320 ears) in non-SSCD group, and the height of the superior semicircular canal was (6.43±0.51)mm, the outer tube diameter was (0.83±0.13)mm, the thickness of the tympanic cavity was (2.19±0.62)mm, the anteroposterior diameter of mastoid was (14.55±1.98)mm; There were 70 cases (113 ears) in the suspicious SSCD group, the above results were (6.42±0.60)mm, (0.85±0.16)mm, (1.62±0.55)mm, (13.24±1.97)mm, respectively. Statistical analysis showed that there were no significant difference in the height of the superior semicircular canal (P=0.94) and the outer tube diameter (P=0.64), There were significant differences in the thickness of the tympanic cavity (P=0.002) and the anteroposterior diameter of mastoid (P=0.004). There were 70 cases (113 ears) in group suspicious SSCD, the unilateral defect had 27 cases (27 ears), and bilateral defect had 43 cases (86 ears). The defect was located in the middle of the parietal wall in 48 ears, located in the anterior wall of 20 ears, located in the posterior wall of 32 ears, more than two defects in 13 ears; 7 ears were mastoid pneumatic type, 90 ears were mastoid sclerotic type, 16 ears were mastoid mixed type.Conclusion:The SSCD was more common in the sclerotic mastoid. The bilateral defect was common, mostly located in the middle of the parietal wall. The occurrence of the lesion is not related to the height and diameter of the superior semicircular canal, but may be related to the thinning of the whole temporal bone.

[Effect of melatonin on expression of Prestin protein in the inner ear of mice following radiotherapy].

Objective: To investigate the effect of melatonin on the expression of prestin protein in the inner ear of mice following a single dose radiation therapy, so as to provide the basis for the mechanism study of radiation induced inner ear injury and its prevention. Methods: Sixty 4-week-old male mice were randomly divided into six groups, including the control group (A group), 50 mg/kg MLT group (B group), 5 mg/kg MLT group (C group), 50 mg/kg MLT + radiotherapy group (D group), 5 mg/kg MLT+ radiotherapy group (E group), and 16 Gy radiotherapy group (F group). Each experimental group was randomly subdivided into two subgroups, which were killed to harvest the cochlea on the 3rd and 7th days following 16 Gy radiation. The specimens were used for immunostaining and Western blot to detect the expression of prestin protein. SPSS 19.0 software was used for statistical analysis. Results: Prestin protein mainly distributed in the lateral membrane above the outer hair cell nucleus. When compared with A, B and C group, the expression of prestin protein in the inner ear was significantly up-regulated in F group (P<0.05). However, D and E group reduced the abnormal expression of prestin following radiotherapy when compared with F group, the difference was statistically significant (P<0.05), and the effect of D group was more significant than E group (P<0.05). Conclusions: The prestin protein of cochlea is mainly distributed in the lateral membrane above the outer hair cell nucleus. Following the high-dose radiotherapy, the prestin expression is upregulated, and melatonin can control the abnormal expression of prestin protein induced by radiotherapy with dose dependent.

Optimisation of breast cancer patients' follow-up - potential way to improve cancer care in transitional countries.

The aim of this analysis was to evaluate adherence of Croatian oncologists to follow-up criteria as suggested by the current national and international guidelines for women with breast cancer receiving adjuvant endocrine therapy. The use of clinical and diagnostic methods was documented in this prospective, non-interventional, multicenter study. A total of 438 post-menopausal patients receiving adjuvant endocrine treatment with non-steroidal aromatase inhibitors were included. Average annual frequency for each clinical and diagnostic method was calculated. Median adjuvant endocrine treatment duration before study recruitment was 10.5 months (interquartile 4.7-26.6). Patients were followed up for an average 23.5 ± 4.9 months. Average number of oncological visits was 5.3. Mammograms were performed at mean annual frequency of 0.7, chest radiographs at 0.5, abdominal ultrasounds at 0.9, breast ultrasounds at 1.2, complete blood counts and chemistry panels at 1.7, carcinoembryonic antigen at 0.8, cancer antigen 15-3 at 1.6, gynaecological examination at 0.3, and densitometry at mean annual frequency of 0.3. In conclusion, among post-menopausal women with breast cancer receiving adjuvant endocrine therapy in this study, more unnecessary and unproven follow-up procedures were done compared to the guidelines' recommendations.

Retrospective analysis of efficacy of trastuzumab in adjuvant treatment of HER 2 positive early breast cancer - single institution experience.

Addition of trastuzumab to chemotherapy is the cornerstone of adjuvant treatment of early HER2 positive breast cancer. Clinical trials and metaanalyses of adjuvant trastuzumab have shown significant reduction in risk of recurrence and death. Nevertheless, the real magnitude of the effect of any drug must be reevaluated in daily clinical conditions, due to the fact that daily clinical practice often differs from conditions in clinical trials. In order to measure the benefit of adding adjuvant trastuzumab in HER 2 positive early breast cancer treatment, we have performed retrospective analysis in a single institution on consecutive patients divided in 2 cohorts: one, treated in "pre - trastuzumab" and the other in "trastuzumab era". Between 2003 and 2012, 258 consecutive HER 2 positive patients with early breast cancer have been treated with adjuvant chemotherapy, 103 patients did not received trastuzumab (patients treated from 2003 till 2007), and 155 (patients treated from 2008 till 2012) received trastuzumab. Patients who received trastuzumab experienced significantly longer median disease-free survival (107 vs. 92 months, LR: 11.6, p <0.001); breast cancer-specific survival (130 vs. 117 months, LR: 10.7, p < 0.001) and median overall survival (123 vs. 108 months LR = 11.6, p < 0.001). The benefits of adding trastuzumab were independent of chemotherapy regimen and hormonal therapy. This retrospective analysis has shown a clear, statistically significant benefit of adjuvant trastuzumab in treatment of early, HER2 positive breast cancer in daily clinical practice, and confirmed the results of the registration clinical trials.

The effects of single-dose dexamethasone on inflammatory response and pain after uterine artery embolisation for symptomatic fibroids or adenomyosis: a randomised controlled study.

OBJECTIVE: To investigate the effects of single-dose intravenous dexamethasone on inflammatory responses, pain, nausea, and vomiting after uterine artery embolisation (UAE). DESIGN: Prospective, randomised, double-blind, and placebo-controlled study. SETTING: Tertiary-care University centre in Korea. POPULATION: Patients undergoing UAE for the treatment of symptomatic fibroids or adenomyosis. METHODS: Patients were randomised to receive either intravenous dexamethasone (10 mg; dexamethasone group) or normal saline (control group) 1 hour before UAE. Both groups received fentanyl-based intravenous patient-controlled analgesia (PCA) during the 24 hours after UAE. MAIN OUTCOME MEASURES: The primary outcomes were the inflammatory and stress responses measured by white blood cell count, neutrophil percentage, C-reactive protein (CRP), interleukin-6 (IL-6), and cortisol. Secondary outcomes were severity of pain and incidence of nausea and vomiting. RESULTS: Sixty-four patients were enrolled and 59 patients completed the study. CRP, IL-6, and cortisol were significantly lower in the dexamethasone group compared with the control group during the 24 hours after UAE. Although the cumulative dose of fentanyl and additional analgesics administered during the 24 hours after UAE were similar between the two groups, pain scores were significantly lower in the dexamethasone group from 12 hours after UAE, and the incidence of severe nausea and vomiting was lower in the dexamethasone group. CONCLUSIONS: The administration of single-dose intravenous dexamethasone as an adjunct to fentanyl-based intravenous PCA is effective in reducing inflammation and pain during the first 24 hours after UAE. TWEETABLE ABSTRACT: Dexamethasone is effective in reducing inflammation and pain after uterine artery embolisation.

Lapatinib in the treatment of HER-2 overexpressing breast cancer.

Lapatinib is the only clinically available agent for the treatment of patients with human epidermal growth factor receptor-2 (HER-2) positive tumors that have progressed on treatment with trastuzumab, taxanes and anthracyclines. Moreover, when given with letrozole in postmenopausal patients with estrogen receptor (ER) and HER-2 positive disease it induces clinically meaningful benefit. Recently presented neoadjuvant data suggests an important place for the combination of trastuzumab and lapatinib in the therapy of early HER-2 positive breast cancer. This article reviews the current status and future perspectives of lapatinib.

A non-synonymous SNP within membrane metalloendopeptidase-like 1 (MMEL1) is associated with multiple sclerosis.

Several single-nucleotide polymorphism (SNP) genome-wide association studies (GWASs) have been completed in multiple sclerosis (MS). Follow-up studies of the variants with the most promising rankings, especially when supplemented by informed candidate gene selection, have proven to be extremely successful. In this study we report the results of a multi-stage replication analysis of the putatively associated SNPs identified in the Wellcome Trust Case Control Consortium non-synonymous SNP (nsSNP) screen. In total, the replication sample consisted of 3444 patients and 2595 controls. A combined analysis of the nsSNP screen and replication data provides evidence implicating a novel additional locus, rs3748816 in membrane metalloendopeptidase-like 1 (MMEL1; odds ratio=1.16, P=3.54 × 10⁻6) in MS susceptibility.

Roles of purines in synaptic modulation evoked by hypercapnia in isolated spinal cord of neonatal rat in vitro.

BACKGROUND AND PURPOSE: The purine compounds, adenosine 5'-triphosphate (ATP) and adenosine, are known to accumulate in the extracellular space and to elicit various cellular responses during hypoxia/ischemia, whereas the roles of purines during hypercapnia are poorly understood. In this study, we examined the effects of various drugs affecting purine turnover on the responses to hypercapnia in the spinal cord. EXPERIMENTAL APPROACH: Electrically evoked reflex potentials were measured in an in vitro preparation of the isolated spinal cord of the neonatal rat by extracellular recording. Extracellular adenosine concentrations were assayed by high performance liquid chromatography (HPLC) methods. KEY RESULTS: Hypercapnia (20% CO2) depressed the reflex potentials, which were partially reversed by an adenosine A1 receptor antagonist, 8-cyclopentyl theophylline, but not by a P2 receptor antagonist, pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid. Exogenous adenosine and ATP also depressed the reflex potentials via adenosine A1 receptors. The hypercapnia-evoked depression was not reversed by inhibitors of gap junction hemichannels, anion channels, P2X7 receptors or equilibrative nucleoside transporters, all of which might be involved in purine efflux pathways. The adenosine accumulation evoked by hypercapnia was not inhibited by tetrodotoxin, ethylene glycol-bis(beta-amino ethyl ether) tetraacetic acid (EGTA) or an ecto-ATPase inhibitor, ARL 67156. Homocysteine thiolactone, used to trap intracellular adenosine, significantly reduced extracellular adenosine accumulation during hypercapnia. CONCLUSIONS AND IMPLICATIONS: These results suggest that hypercapnia released adenosine itself from intracellular sources, using pathways different from the conventional exocytotic mechanism, and that this adenosine depressed spinal synaptic transmission via adenosine A1 receptors.

The 2.6 A resolution structure of Rhodobacter capsulatus bacterioferritin with metal-free dinuclear site and heme iron in a crystallographic 'special position'.

Bacterioferritin from Rhodobacter capsulatus was crystallized and its structure was solved at 2.6 A resolution. This first structure of a bacterioferritin from a photosynthetic organism is a spherical particle of 24 subunits displaying 432 point-group symmetry like ferritin and bacterioferritin from Escherichia coli. Crystallized in the I422 space group, its structural analysis reveals for the first time the non-symmetric heme molecule located on a twofold crystallographic symmetry axis. Other hemes of the protomer are situated on twofold noncrystallographic axes. Apparently, both types of sites bind heme in two orientations, leading to an average structure consisting of a symmetric 50:50 mixture, thus satisfying the crystallographic and noncrystallographic symmetry of the crystal. Five water molecules are situated close to the heme, which is bound in a hydrophobic pocket and axially coordinated by two crystallographic or noncrystallographically related methionine residues. Its ferroxidase center, in which Fe(II) is oxidized to Fe(III), is empty or fractionally occupied by a metal ion. Two positions are observed for the coordinating Glu18 side chain instead of one in the E. coli enzyme in which the site is occupied. This result suggests that the orientation of the Glu18 side chain could be constrained by this interaction.

A refractory case of secondary erythermalgia successfully treated with lumbar sympathetic ganglion block.

A 59-year-old Japanese man with myasthenia gravis, who had a 10-year history of temperature-sensitive pain in the lower extremities, i.e. improved by cooling and worsened by warming, consulted us because the pain had become intolerable during the previous 4 months. Bilateral erythema, swelling and large ulcers were noted on the calves, dorsal aspects of the feet, and soles. Laboratory data showed thrombocythaemia and a positive antibody to the acetylcholine receptor, but were negative for antinuclear and antiphospholipid antibodies. A diagnosis of secondary erythermalgia was made because of the clinical features, the laboratory data, and the lack of family history of this disease. Although steroid pulse therapy, oral aspirin and antiserotonin drugs were ineffective, bilateral lumbar sympathetic ganglion block succeeded in relieving the severe pain and curing the ulcers. The clinical course in our patient suggests that sympathetic ganglion block may be one of the most effective treatments for secondary erythermalgia. Although the mechanism of this effect is uncertain, microcirculation disturbance in secondary erythermalgia, if any, may be improved by this block.

Tufted angioma of adult onset, revealing abundant eccrine glands and central regression.

Tufted angioma (angioblastoma) usually occurs in infancy. Some lesions reveal proliferation of eccrine glands and some undergo spontaneous regression. We report a lesion from the left lateral chest of a 58-year-old male, which had appeared 5 years ago and gradually increased in size. Its color had faded out centrally. Histological examination showed many angiomatous lobules on the border of the lesion but few in the central portion. Abundant eccrine sweat glands, small venules and dense connective tissue were seen in both areas. The histology and the clinical course suggested that central regression occurred in the lesion. Out of 211 reported cases, we found 4 cases that had annular lesions spreading centrifugally. Though these lesions were not examined histologically, they were suspected to regress in the central portions. We suggest that central regression can rarely occur in tufted angioma.

Infection-susceptibility alleles of mannose-binding lectin are associated with increased carotid plaque area.

The MBL gene, encoding mannose-binding lectin, determines interindividual variation in susceptibility to certain infectious agents, such as Chlamydia pneumoniae. We examined whether infection-susceptibility alleles of MBL, called "non-A alleles," would be associated with increased carotid plaque area (CPA), an intermediate phenotype of atherosclerosis. In 164 subjects, we measured CPA with 2-dimensional ultrasound. We also determined traditional atherosclerosis risk factors and genotyped all subjects for MBL codons 52, 54, and 57. We used ANOVA to determine sources of variation for CPA and tested the hypothesis that the presence of a single MBL non-A "infection-susceptibility" allele was associated with increased CPA; 45.7% of subjects had at least one non-A allele. ANOVA showed that CPA was significantly associated with MBL genotype, age, smoking, hypertension, and hyperlipidemia (P < 0.05). When MBL was used as the sole independent variable in the regression analysis, the association with CPA was even more significant (P = 0.009). Subjects with at least one MBL non-A allele had significantly higher CPA than subjects homozygous for the MBL A allele and were significantly more likely to have CPA in excess of the sample median. Thus, infection-susceptibility alleles of MBL were associated with increased CPA in this study sample; these alleles may be a determinant of interindividual differences in atherosclerosis risk.

Role of the cyclic AMP response element binding complex and activation of mitogen-activated protein kinases in synergistic activation of the glycoprotein hormone alpha subunit gene by epidermal growth factor and forskolin.

The aim of these studies was to elucidate a role for epidermal growth factor (EGF) signaling in the transcriptional regulation of the glycoprotein hormone alpha subunit gene, a subunit of chorionic gonadotropin. Studies examined the effects of EGF and the adenylate cyclase activator forskolin on the expression of a transfected alpha subunit reporter gene in a human choriocarcinoma cell line (JEG3). At maximal doses, administration of EGF resulted in a 50% increase in a subunit reporter activity; forskolin administration induced a fivefold activation; the combined actions of EGF and forskolin resulted in synergistic activation (greater than eightfold) of the alpha subunit reporter. Mutagenesis studies revealed that the cyclic AMP response elements (CRE) were required and sufficient to mediate EGF-forskolin-induced synergistic activation. The combined actions of EGF and forskolin resulted in potentiated activation of extracellular signal-regulated kinase (ERK) enzyme activity compared with EGF alone. Specific blockade of ERK activation was sufficient to block EGF-forskolin-induced synergistic activation of the alpha subunit reporter. Pretreatment of JEG3 cells with a p38 mitogen-activated protein kinase inhibitor did not influence activation of the alpha reporter. However, overexpression of c-Jun N-terminal kinase (JNK)-interacting protein 1 as a dominant interfering molecule abolished the synergistic effects of EGF and forskolin on the alpha subunit reporter. CRE binding studies suggested that the CRE complex consisted of CRE binding protein and EGF-ERK-dependent recruitment of c-Jun-c-Fos (AP-1) to the CRE. A dominant negative form of c-Fos (A-Fos) that specifically disrupts c-Jun-c-Fos DNA binding inhibited synergistic activation of the alpha subunit. Thus, synergistic activation of the alpha subunit gene induced by EGF-forskolin requires the ERK and JNK cascades and the recruitment of AP-1 to the CRE binding complex.

Effects of nickel sulfate on pulmonary natural immunity in Wistar rats.

This study was designed to investigate the in vivo effect of nickel sulfate on the pulmonary non-specific immune defences. Groups of four male Wistar rats were treated with a single intratracheal instillation of NiSO(4) at different doses: 1, 2, 4 and 8 micromol of NiSO(4) per rat. Control rats received a corresponding instillation of the saline vehicle. The effect of NiSO(4) on the cytotoxic activity of the pulmonary natural killer (NK) cells and alveolar macrophages (AM), as well as the pulmonary production of cytokines such as alpha-tumor necrosis factor (TNF-alpha) and gamma-interferon (IFN-gamma), were examined 1, 2 and 7 days later. Spontaneous NK-cytotoxicity towards mouse-derived tumor cell line, Yac-1 was suppressed 1 day after treatment at doses of 2 micromol/rat and above with only one result significant (P<0.05); 2 days after treatment the suppression was increased with all results significant at the same doses; 1 week after treatment NK activity restoration was observed except for the highest dose, 8 micromol/rat. AM-mediated cytotoxicity towards mouse-derived tumor cell line, 3T12, did not show any significant difference in treated and untreated animals. In contrast, whereas moderate levels of TNF-alpha were detected in the broncho-alveolar lavage (BAL) fluid supernatants of controls, the NiSO(4) treatment highly suppressed TNF-alpha production with a maximum observed after 2 days. TNF-alpha suppression was found to be transient, at least with the lowest NiSO(4) dose, with levels returning to normal after 7 days. A non-significant increase in IFN-gamma was observed in the BAL fluids of treated animals at each time of examination. Taken together, these results indicate that NK cell activity and TNF-alpha secretion are sensitive targets for instilled NiSO(4) in Wistar rats.

Fibroblast growth factor-2.

Fibroblast growth factor-2 (FGF-2) is a heparin-binding growth factor which occurs in several isoforms resulting from alternative initiations of translation: an 18 kD cytoplasmic isoform and four larger molecular weight nuclear isoforms (22, 22.5, 24 and 34 kD). FGF-2 has pleiotropic roles in many cell types and tissues; it is a motogenic, angiogenic and survival factor which is involved in cell migration, cell differentiation and in a variety of developmental processes. Although devoid of signal peptide, it could be secreted. It acts mainly through a paracrine/autocrine mechanism involving high affinity transmembrane receptors and heparan sulfate proteoglycan low affinity receptors, but also through still unknown intracrine process(es) on intracellular targets. FGF-2 has many biological functions which are probably isoform-specific. Nevertheless, FGF-2 is not essential for embryonic development as knock-out mice for the growth factor are viable and fertile although they exhibit abnormalities in neuronal differentiation. Use of FGF-2 as therapeutic agent for the treatment of ischemic cardiovascular disease is promising and clinical trials are in progress.

Nuclear 24 kD fibroblast growth factor (FGF)-2 confers metastatic properties on rat bladder carcinoma cells.

The tumorigenic and metastatic properties of rat bladder carcinoma NBT-II cells transfected with a cDNA encoding the 24 kD nuclear isoform of human fibroblast growth factor-2 (FGF-2) were analysed and compared with those cells producing the 18 kD cytoplasmic isoform FGF-2. In transfected clones, 24 kD FGF-2 was found in the nucleus, and no FGF-2 was secreted. RT-PCR analysis showed no upregulation of FGF-2-specific receptor FGFR2c expression in these proliferating transfected cells. A shorter latency period for in vivo tumor formation and abundant spontaneous lung metastases were only seen if nuclear FGF-2-producing cells were injected subcutaneously into nude mice. Intravenous injection of 24 kD FGF-2-producing cells led to extensive experimental lung metastases whereas injection of control NBT-II cells or 18 kD FGF-2-producing cells did not. As FGF-2-producing cells have no specific FGF-2 receptors, our results suggest that the 24 kD FGF-2 has nuclear targets, and activates metastatic property of carcinoma cells via a mechanism other than the conventional FGF receptor-mediated signaling pathway.

[Impact on tumor angiogenesis and tumor progression of expression of the 18 kd and 24 kd isoforms of FGF-2]. / Impact sur l'angiogenèse tumorale et la progression tumorale de l'expression des isoformes 18 kD et 24 kD du FGF-2.

The role of FGF-2 in tumor progression and tumor cell invasiveness was investigated using the rat bladder carcinoma cells NBT-II, which do not constitutively express FGF-2 or its membrane-spanning receptor. The NBT-II cells were transfected using expression vectors encoding either the 18 kD or the 24 kD isoform of FGF-2. The 24 kD isoform contains a nuclear localization signal. The transfected NBT-II cells that expressed 18 kD FGF-2 produced and secreted this factor as the biologically active form and retained an epithelial morphology. When injected to nude mice, the tumorigenic potential of these cells was not increased over that of non-transfected NBT-II cells; however, although the time to tumor development was long, the tumors were highly vascularized, indicating secretion of the angiogenic factor FGF-2. The transfected NBT-II cells that expressed 24 kD FGF-2 varied in their morphological appearance and did not secrete FGF-2; immunofluorescence and Western-blot studies showed that the FGF-2 was mainly intranuclear. When injected to nude mice, these cells produced tumors and migrated not only to the lymph nodes but also to the lungs where they produced metastases. In aggregate, these data indicate that stimulation of angiogenesis is not sufficient to increase tumor growth and that nuclear FGF-2 acts as a tumorigenic and metastasis-promoting factor in the NBT-II carcinoma model.

Role of SAM-dependent thiol methylation in the renal toxicity of several solvents in mice.

The role of S-adenosylmethionine (SAM)-dependent thiol methylation in the nephrotoxicity of seven industrial solvents was studied in mice. The seven following solvents were utilized: bromobenzene (BB), styrene (STY), tetrachloroethylene (TTCE), trichloroethylene (TCE), 1,1-dichloroethylene (DCE), 1,2-dichloroethane (DCA) and hexachlorobutadiene (HCB). The experimental model comprised mice pretreated with periodate oxidized adenosine (ADOX) (100 micromol kg(-1) i.p.) 30 min before injection of solvents. In the first 4 h after ADOX treatment, the SAM levels were about fourfold higher than controls for the liver and kidney. The S-adenosylhomocysteine (SAH) levels were increased by factors of 11 and 14 and the SAM/SAH ratios were decreased by factors of 3 and 10 for the liver and kidney, respectively. These results show that ADOX treatment probably induces an inhibition of methyltransferase SAM-dependent in the liver and kidney and thus decreases the methylation capabilities. A single oral administration of BB (500 or 800 mg kg(-1)), TTCE (3500 or 4000 mg kg(-1)), TCE (3000 or 3500 mg kg(-1)) or STY (400 or 600 mg kg(-1)) did not induce renal toxicity, evaluated by the percentage of damaged tubules compared to controls. On the other hand, the three solvents DCE, HCB and DCA were nephrotoxic and the percentage of damaged tubules observed for each solvent was significantly different from the value of <1.8% for controls: 19% and 40% for DCE (130 and 200 mg kg(-1)), 50% and 46% for HCB (80 and 100 mg kg(-1)) and 5.1% and 7.6% for DCA (1000 and 1500 mg kg(-1)). The ADOX treatment in the mice did not modify the renal toxicity of the seven solvents. Thus, their renal toxicity, when it existed, was probably independent of the SAM-dependent thiolmethyltransferase activity in the mice. The results of this study are discussed from two viewpoints. The first concerns the general considerations on inhibition of thiol methyltransferase activities in mice and the second is related to the different solvents that are evoked individually.

Lipoprotein-genotype associations in Trinidadian neonates.

OBJECTIVES: We hypothesized that common variation in the angiotensinogen (AGT), beta-3-adrenergic receptor, intestinal fatty acid-binding protein, serum paraoxonase, paraoxonase-2, hepatic lipase, apolipoprotein E (APOE), and Werner helicase (WRN) genes would be associated with variation in biochemical phenotypes in a previously unstudied neonatal sample. DESIGN AND METHODS: We examined associations of both nongenetic and genetic variables with plasma lipoprotein traits in neonates from Trinidad. RESULTS: Among nongenetic variables, we found significant associations between plasma concentrations of: 1) lipoprotein(a) [Lp(a)] and both ethnicity (p = 0.037) and birth weight (p = 0.001); 2) total cholesterol and gender (p = 0.010); 3) triglyceride and birth weight (p = 0.035); and 4) apolipoprotein AI and gender (p = 0.016). Among genetic variables, we found that: 1) common variation on chromosome 1q in AGT codon 235 was significantly associated with variation in plasma apolipoproteins AI (p<0.0001) and B (p = 0.013); 2) common variation in WRN at codon 1367 was significantly associated with variation in plasma Lp(a) (p<0.0001); and 3) common variation in APOE at codons 112 and 158 was significantly associated with variation in plasma triglycerides (p = 0.013). CONCLUSIONS: The associations with AGT and WRN are novel and may have resulted either from a direct influence of the genetic variants or through linkage disequilibrium with other functional loci, such as the familial combined hyperlipidemia locus on chromosome 1q in the case of AGT. Despite the fact that there are some limitations in making determinations from cord blood, the results suggest that there may be genetic determinants of plasma lipoproteins in neonates.