The Rabbit Phosphofructokinase Genes: Molecular Cloning, Sequencing and Characterization.

Two rabbit phosphofructokinase (PFK) genomic clones were isolated from a lambda Charon 4A rabbit liver DNA library. A 180-bp rabbit muscle PFK cDNA encoding 60 amino acids in the C-terminal region of RM-PFK was used as a probe. By using the shotgun subcloning approach, we cloned, sequenced and characterized two rabbit PFK genomic clones. The two clones overlapped and represented a total length of 17 kb which encoded the entire RM-PFK protein. The organization of this gene was elucidated by restriction mapping and DNA sequencing. It contains 22 exons, ranging in size from 45 to 190 bp, separated by 21 introns of 78-3,500 in length. This gene can be divided into two nearly homologous halves, the N- and C-halves. Each half is composed of almost an equal number of exons. The 22 exons code for 779 amino acid residues with a calculated molecular mass of 84,975 daltons. Exons XV and XVI together code for the 30 amino acid residues which were left as an unidentified gap in the published primary structure for this enzyme. Sequence analysis showed that 74% of the bases at the third position of the codons in the coding exons are either G or C. When overlaid on the amino acid sequence of the protein, most of the introns are located between or near the ends of the secondary structural elements. However, introns are not located at analogous positions in the two protein-coding halves of the gene. Sequence homologies between bacterial and rabbit muscle PFKs and between the amino- and carboxyl-terminal halves of the latter suggest that the mammalian enzyme evolved from a prokaryotic progenitor by gene duplication and divergence. A cDNA clone encoding one-quarter of the RM-PFK sequence was sequenced. The nucleotide sequence of the cDNA agrees with the determined RM-PFK genomic sequence. In addition, the amino acid sequence deduced from the cDNA is identical to the RM-PFK sequence previously reported by Poorman et al

Hyperbaric oxygen upregulates cochlear constitutive nitric oxide synthase

<p>Abstract</p> <p>Background</p> <p>Hyperbaric oxygen therapy (HBOT) is a known adjuvant for treating ischemia-related inner ear diseases. Controversies still exist in the role of HBOT in cochlear diseases. Few studies to date have investigated the cellular changes that occur in inner ears after HBOT. Nitric oxide, which is synthesized by nitric oxide synthase (NOS), is an important signaling molecule in cochlear physiology and pathology. Here we investigated the effects of hyperbaric oxygen on eardrum morphology, cochlear function and expression of NOS isoforms in cochlear substructures after repetitive HBOT in guinea pigs.</p> <p>Results</p> <p>Minor changes in the eardrum were observed after repetitive HBOT, which did not result in a significant hearing threshold shift by tone burst auditory brainstem responses. A differential effect of HBOT on the expression of NOS isoforms was identified. Upregulation of constitutive NOS (nNOS and eNOS) was found in the substructures of the cochlea after HBOT, but inducible NOS was not found in normal or HBOT animals, as shown by immunohistochemistry. There was no obvious DNA fragmentation present in this HBOT animal model.</p> <p>Conclusions</p> <p>The present evidence indicates that the customary HBOT protocol may increase constitutive NOS expression but such upregulation did not cause cell death in the treated cochlea. The cochlear morphology and auditory function are consequently not changed through the protocol.</p

Staggered intercalation of DNA duplexes with base-pair modulation by two distinct drug molecules induces asymmetric backbone twisting and structure polymorphism

The use of multiple drugs simultaneously targeting DNA is a promising strategy in cancer therapy for potentially overcoming single drug resistance. In support of this concept, we report that a combination of actinomycin D (ActD) and echinomycin (Echi), can interact in novel ways with native and mismatched DNA sequences, distinct from the structural effects produced by either drug alone. Changes in the former with GpC and CpG steps separated by a A:G or G:A mismatch or in a native DNA with canonical G:C and C:G base pairs, result in significant asymmetric backbone twists through staggered intercalation and base pair modulations. A wobble or Watson-Crick base pair at the two drug-binding interfaces can result in a single-stranded 'chair-shaped' DNA duplex with a straight helical axis. However, a novel sugar-edged hydrogen bonding geometry in the G:A mismatch leads to a 'curved-shaped' duplex. Two non-canonical G:C Hoogsteen base pairings produce a sharply kinked duplex in different forms and a four-way junction-like superstructure, respectively. Therefore, single base pair modulations on the two drug-binding interfaces could significantly affect global DNA structure. These structures thus provide a rationale for atypical DNA recognition via multiple DNA intercalators and a structural basis for the drugs' potential synergetic use

Well-differentiated gall bladder hepatoid carcinoma producing alpha-fetoprotein: a case report

<p>Abstract</p> <p>Introduction</p> <p>Gall bladder carcinoma is rare, and metastatic gall bladder carcinoma from hepatocellular carcinoma has been reported in only a few patients.</p> <p>Case presentation</p> <p>We present a 73-year-old man with a history of hepatitis B virus-related liver cirrhosis and hepatocellular carcinoma. He received transcatheter arterial chemoembolization, and was diagnosed to have an alpha-fetoprotein producing gall bladder tumor with intraluminal growth. Open cholecystectomy was performed. Pathologic examination of the lesion revealed a well-differentiated hepatoid carcinoma. The lesion was thought most likely to be a metastatic lesion from previous hepatocellular carcinoma. His alpha-fetoprotein level dropped to normal levels five months after the surgery.</p> <p>Conclusion</p> <p>This unusual intraluminal growing tumor proved to be a well-differentiated hepatoid carcinoma, most likely a metastatic lesion from previous hepatocellular carcinoma. This case reminds clinicians that in looking for likely hepatocellular carcinoma recurrence, when no detectable hepatic lesion can account for an elevated alpha-fetoprotein level, the gall bladder should be included in the search for the site of metastasis.</p

Developing the Effective Method of Spectral Harmonic Energy Ratio to Analyze the Arterial Pulse Spectrum

In this article, we analyze the arterial pulse in the spectral domain. A parameter, the spectral harmonic energy ratio (SHER), is developed to assess the features of the overly decreased spectral energy in the fourth to sixth harmonic for palpitation patients. Compared with normal subjects, the statistical results reveal that the mean value of SHER in the patient group (57.7 ± 27.9) is significantly higher than that of the normal group (39.7 ± 20.9) (P-value = .0066 < .01). This means that the total energy in the fourth to sixth harmonic of palpitation patients is significantly less than it is in normal subjects. In other words, the spectral distribution of the arterial pulse gradually decreases for normal subjects while it decreases abruptly in higher-order harmonics (the fourth, fifth and sixth harmonics) for palpitation patients. Hence, SHER is an effective method to distinguish the two groups in the spectral domain. Also, we can thus know that a “gradual decrease” might mean a “balanced” state, whereas an “abrupt decrease” might mean an “unbalanced” state in blood circulation and pulse diagnosis. By SHER, we can determine the ratio of energy distribution in different harmonic bands, and this method gives us a novel viewpoint from which to comprehend and quantify the spectral harmonic distribution of circulation information conveyed by the arterial pulse. These concepts can be further applied to improve the clinical diagnosis not only in Western medicine but also in traditional Chinese medicine (TCM)

Pulsed laser deposition of hexagonal GaN-on-Si(100) template for MOCVD applications

Growth of hexagonal GaN on Si(100) templates via pulsed laser deposition (PLD) was investigated for the further development of GaN-on-Si technology. The evolution of the GaN growth mechanism at various growth times was monitored by SEM and TEM, which indicated that the GaN growth mode changes gradually from island growth to layer growth as the growth time increases up to 2 hours. Moreover, the high-temperature operation (1000°C) of the PLD meant no significant GaN meltback occurred on the GaN template surface. The completed GaN templates were subjected to MOCVD treatment to regrow a GaN layer. The results of X-ray diffraction analysis and photoluminescence measurements show not only the reliability of the GaN template, but also the promise of the PLD technique for the development of GaN-on-Si technology

A 64-week, multicenter, open-label study of aripiprazole effectiveness in the management of patients with schizophrenia or schizoaffective disorder in a general psychiatric outpatient setting

<p>Abstract</p> <p>Objective</p> <p>To evaluate the overall long-term effectiveness of aripiprazole in patients with schizophrenia in a general psychiatric practice setting in Taiwan.</p> <p>Methods</p> <p>This was a prospective, open-label, multicenter, post-market surveillance study in Taiwanese patients with a Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) diagnosis of schizophrenia or schizoaffective disorder requiring a switch in antipsychotic medication because current medication was not well tolerated and/or clinical symptoms were not well controlled. Eligible patients were titrated to aripiprazole (5-30 mg/day) over a 12-week switching phase, during which their previous medication was discontinued. Patients could then enter a 52-week, long-term treatment phase. Aripiprazole was flexibly dosed (5-30 mg/day) at the discretion of the treating physicians. Efficacy was assessed using the Clinical Global Impression scale Improvement (CGI-I) score, the Clinical Global Impression scale Severity (CGI-S) score, The Brief Psychiatry Rating Scale (BPRS), and the Quality of Life (QOL) scale, as well as Preference of Medicine (POM) ratings by patients and caregivers. Safety and tolerability were also assessed.</p> <p>Results</p> <p>A total of 245 patients were enrolled and switched from their prior antipsychotic medications, and 153 patients entered the 52-week extension phase. In all, 79 patients (32.2%) completed the study. At week 64, the mean CGI-I score was 3.10 and 64.6% of patients who showed response. Compared to baseline, scores of CGI-S, QOL, and BPRS after 64 weeks of treatment also showed significant improvements. At week 12, 65.4% of subjects and 58.9% of caregivers rated aripiprazole as better than the prestudy medication on the POM. The most frequently reported adverse events (AEs) were headache, auditory hallucinations and insomnia. A total of 13 patients (5.3%) discontinued treatment due to AEs. No statistically significant changes were noted with respect to fasting plasma glucose, lipid profile, body weight, and body mass index after long-term treatment with aripiprazole.</p> <p>Conclusions</p> <p>Although the discontinuation rate was high, aripiprazole was found to be effective, safe and well tolerated in the long-term treatment of Taiwanese patients with schizophrenia who continued to receive treatment for 64 weeks.</p

Radial Pressure Pulse and Heart Rate Variability in Heat- and Cold-Stressed Humans

This study aims to explore the effects of heat and cold stress on the radial pressure pulse (RPP) and heart rate variability (HRV). The subjects immersed their left hand into 45°C and 7°C water for 2 minutes. Sixty healthy subjects (age 25 ± 4 yr; 29 men and 31 women) were enrolled in this study. All subjects underwent the supine temperature measurements of the bilateral forearms, brachial arterial blood pressure, HRV and RPP with a pulse analyzer in normothermic conditions, and thermal stresses. The power spectral low-frequency (LF) and high-frequency (HF) components of HRV decreased in the heat test and increased in the cold test. The heat stress significantly reduced radial augmentation index (AIr) (P < .05), but the cold stress significantly increased AIr (P < .01). The spectral energy of RPP did not show any statistical difference in 0 ~ 10 Hz region under both conditions, but in the region of 10 ~ 50 Hz, there was a significant increase (P < .01) in the heat test and a significant decrease in the cold test (P < .01). The changes in AIr induced by heat and cold stress were significantly negatively correlated with the spectral energy in the region of 10 ~ 50 Hz (SE10−50 Hz) but not in the region of 0 ~ 10 Hz (SE0−10 Hz). The results demonstrated that the SE10−50 Hz, which only possessed a small percentage in total pulse energy, presented more physiological characteristics than the SE0−10 Hz under the thermal stresses