Odor detection ability and thallium-201 transport in the olfactory nerve of traumatic olfactory-impaired mice

Although olfactory nerve damage is a contributing factor in the diagnosis of posttraumatic olfactory loss, at present, there are no methods to directly assess injury to these nerves. We have shown that following olfactory nerve injury in mice, thallium-201 (201 Tl) transport from the nasal cavity to the olfactory bulb decreases. To determine if olfactory function after nerve injury could be assessed with nasal administration of 201 Tl, we measured the correlation between odor detection ability (ODA) and the rate of transport of 201 Tl in olfactory nerves. Both ODA and 201 Tl transport were measured after bilateral olfactory nerve transection for a 4-week period. Cycloheximide solution was used for ODA against tap water. 201 Tl transport was measured as the ratio of radioactivity in the nasal cavity and olfactory bulb with gamma spectrometry. There was a significant correlation between ODA and the rate of 201 Tl transport in the olfactory nerve. These findings suggest that olfactory function after nerve injury can be objectively evaluated with the nasal administration of 201 Tl.     

Identification of a WT1 protein‐derived peptide,WT1187, as a HLA‐A*0206‐restricted,WT1‐specific CTL epitope

The Wilms' tumor gene WT1 is overexpressed in various kinds of hematopoietic malignancies as well as solid cancers, and this protein has been demonstrated to be an attractive target antigen for cancer immunotherapy. WT1‐specific CTL epitopes with a restriction of HLA‐A*2402 or HLA‐A*0201 have been already identified. In the present study it has been demonstrated that a 9‐mer WT1‐derived WT1₁₈₇ peptide, which had already been shown to elicit a WT1‐specific CTL response with a restriction of HLA‐A*0201, can also elicit a CTL response with a restriction of HLA‐A*0206. In all three different HLA‐A*0206⁺ healthy donors examined, WT1₁₈₇ peptide‐specific CTL could be generated from peripheral blood mononuclear cells, and the CTL showed cytotoxic activity that depended on dual expression of WT1 and HLA‐A*0206 molecules. The present study describes the first identification of a HLA‐A*0206‐restricted, WT1‐specific CTL epitope. The present results should help to broaden the application of WT1 peptide‐based immunotherapy from only HLA‐A*0201‐positive to HLA‐A*0206‐positive cancer patients as well.     

Effectiveness of paroxetine in the treatment of acute major depression in adults: a systematic re-examination of published and unpublished data from randomized trials

Background

Concern has been raised about the efficacy of antidepressant therapy for major depression in adults. We undertook a systematic review of published and unpublished clinical trial data to determine the effectiveness and acceptability of paroxetine.

Methods

We searched the Cochrane Collaboration Depression, Anxiety and Neurosis Controlled Trials Register, the Cochrane Central Register of Controlled Trials, the GlaxoSmithKline Clinical Trial Register, MEDLINE and EMBASE up to December 2006. Published and unpublished randomized trials comparing paroxetine with placebo in adults with major depression were eligible for inclusion. We selected the proportion of patients who left a study early for any reason as the primary outcome measure because it represents a hard measure of treatment effectiveness and acceptability.

Results

We included in our review 29 published and 11 unpublished clinical trials, with a total of 3704 patients who received paroxetine and 2687 who received with placebo. There was no difference between paroxetine and placebo in terms of the proportion of patients who left the study early for any reason (random effect relative risk [RR] 0.99, 99% confidence interval [CI] 0.88–1.11). Paroxetine was more effective than placebo, with fewer patients who did not experience improvement in symptoms of at least 50% (random effect RR 0.83, 99% CI 0.77–0.90). Significantly more patients in the paroxetine group than in the placebo group left their respective studies because of side effects (random effect RR 1.77, 95% CI 1.44–2.18) or experienced suicidal tendencies (odds ratio 2.55, 95% CI 1.17–5.54).

Interpretation

Among adults with moderate to severe major depression in the clinical trials we reviewed, paroxetine was not superior to placebo in terms of overall treatment effectiveness and acceptability. These results were not biased by selective inclusion of published studies.Although most treatment guidelines recommend one of the selective serotonin reuptake inhibitors in the pharmacologic treatment of moderate to severe depression in adults,^1,2 concerns have been raised in recent years about the efficacy of these drugs in alleviating symptoms of depression.^3,4First, in trials of antidepressants, the choice of the outcome of interest is often problematic. Whereas in other fields of medicine the definition of outcome measures may be relatively straightforward, efficacy in the treatment of depression may be an elusive concept, typically measured by rating scales. The use of rating scales as outcome measures has often been questioned by physicians, who seldom use them to define patients'' improvement under clinical circumstances.⁵ In addition, improvement in symptoms is typically documented as the difference between baseline and post-treatment scores. Although from a practical viewpoint this approach seems reasonable, in that it allows physicians to make a reasoned judgment in terms of proportions of patients (and not in terms of means and standard deviations), it may systematically magnify the effect of new drugs relative to placebo.³ As a consequence of this criticism, the field of mental health has seen a renewal of interest in “hard measures” of treatment effectiveness.^6–8 Hard measures include suicide attempts, treatment switching, hospital admissions, job loss or dropping out of the trial itself. Hard outcomes may also have a role in re-analyses of clinical trial data, where they may offer a “down-to-earth” evaluation of effectiveness and acceptability.A second concern is that the modest differences between active antidepressants and placebo, as calculated in systematic reviews of clinical trial data,^9–12 might be explained in part by the selective inclusion of specific subsets of studies, such as those submitted to regulatory authorities by drug companies, those that were eventually published or those supported by the manufacturers of selective serotonin reuptake inhibitors.¹³In the present systematic review, we used a hard measure to determine the effectiveness and acceptability of paroxetine, a commonly prescribed antidepressant belonging to the group of selective serotonin reuptake inhibitors. Paroxetine was chosen for 3 reasons: first, it is one of the most frequently prescribed antidepressant drugs both in primary and secondary care (as indicated by a search of the US National Center for Health Statistics database, www2.cdc.gov/drugs/); second, GlaxoSmithKline, the company that launched paroxetine, has recently adopted and implemented a disclosure policy to provide easy access to all published and unpublished data from clinical trials that it has sponsored; and third, the effectiveness and acceptability of paroxetine are particularly relevant in view of recent concerns about its safety profile, especially in terms of suicidal tendencies among pediatric and adult patients with major depression.^14–17     

The looped heart does not save energy by maintaining the momentum of blood flowing in the ventricle

Previous studies suggested that the reconstruction or maintenance of physiological blood flow paths in the heart is important to obtain a good outcome following cardiac surgery, but this concept has no established theoretical foundation. We developed a multiscale, multiphysics heart simulator, based on the finite element method, and compared the hemodynamics of ventricles with physiological and nonphysiological flow paths. We found that the physiological flow path did not have an energy-saving effect but facilitated the separation of the outflow and inflow paths, so avoiding any mixing of the blood. The work performed by the ventricular wall was comparable at slower and faster heart rates (physiological vs. nonphysiological, 0.864 vs. 0.874 J, heart rate = 60 beats/min; and 0.599 vs. 0.590 J, heart rate = 100 beats/min), indicating that chiral asymmetry of the flow paths in the mammalian heart has minimal functional merit. At lower heart rates, the blood coming in the first beat was cleared almost completely by the ninth beat in both models. However, at high heart rates, such complete clearance was observed only in the physiological model, whereas 27.0% of blood remained in the nonphysiological model. This multiscale heart simulator provided detailed information on the cardiac mechanics and flow dynamics and could be a useful tool in cardiac physiology.     

Establishment and characterization of a cell line (IGSK-3) secreting human chorionic gonadotropin, adrenocorticotropic hormone and parathyroid hormone-related protein derived from primary poorly differentiated adenocarcinoma of the stomach

We recently established human chorionic gonadotropin-, adrenocorticotropic hormone- and parathyroid hormone-related protein-secreting cell line derived from primary poorly differentiated adenocarcinoma of the stomach. The cell line was designated as IGSK-3. Inverted-phase contrast microscopy revealed that the IGSK-3 cells consist of two morphological subtypes. One type has visible nucleoli and clear nuclei, but nucleoli and nuclear membrane of the other type are invisible. The population-doubling time was about 43 h. An analysis of conditioned medium by IGSK-3 cells cultured for 4 days revealed the IGSK-3 cells secrete human chorionic gonadotropin-beta (0.5 ng/mL), adrenocorticotropic hormone (5.5 pg/mL), parathyroid hormone-related protein (3.4 pmol/mL) and epidermal growth factor (14.2 pg/mL). Histopathological diagnosis of the graft of IGSK-3 cells revealed that IGSK-3 cells built a poorly differentiated adenocarcinoma which resembled the original tumor. In addition, the IGSK-3 cell line was immunocytochemically positive for human chorionic gonadotropin-beta and epidermal growth factor receptor, and negative for vascular endothelial growth factor.     

Diversity and similarity of microbial communities in petroleum crude oils produced in Asia

To understand microbial communities in petroleum crude oils, we precipitated DNA using high concentrations of 2,2,4-trimethylpentane (isooctane) and purified. Samples of DNA from five crude oils, (Middle East, 3; China, 1; and Japan, 1) were characterized based upon their 16S rRNA gene sequences after PCR amplification and the construction of clone libraries. We detected 48 eubacterial species, one cyanobacterium, and one archaeon in total. The microbial constituents were diverse in the DNA samples. Most of the bacteria affiliated with the sequences of the three oils from the Middle East comprised similar mesophilic species. Acinetobacter, Propionibacterium, Sphingobium and a Bacillales were common. In contrast, the bacterial communities in Japanese and Chinese samples were unique. Thermophilic Petrotoga-like bacteria (11%) and several anaerobic-thermophilic Clostridia- and Synergistetes-like bacteria (20%) were detected in the Chinese sample. Different thermophiles (12%) and Clostridia (2%) were detected in the Japanese sample.     

Adsorption of humic acid from aqueous solution onto irradiation-crosslinked carboxymethylchitosan

This paper presents the adsorption of humic acid from aqueous solution onto crosslinked chitosan derivative (carboxymethylchitosan), formed by additionless irradiation technique. The surface charge and swelling properties of crosslinked samples were investigated. The adsorption of humic acid onto crosslinked carboxymethylchitosan was carried out by the batch method at room temperature, and it was found to be strongly pH-dependent. Maximum amount of humic acid was adsorbed under acidic conditions at the optimum pH value of 3.5. Adsorption kinetic studies indicated the adsorption process was transport-limited at the same pH. The adsorption isotherm analysis data under various initial humic acid concentrations confirms that experimental data fitted well into the Langmuir equation. X-ray photoelectron spectroscopy (XPS) revealed that the amino groups of carboxymethylated chitosan were protonated, suggesting the formation of organic complex between the protonated amino groups and humic acid. From these preliminary evaluations, it was concluded that crosslinked carboxymethylated chitosan derivatives have a great potential in water treatment for the removal of humic acid and other polarized or electrically charged species.     

The mechanism selecting the guide strand from small RNA duplexes is different among argonaute proteins

Double-stranded RNA induces RNA silencing and is cleaved into21–24 nt small RNA duplexes by Dicer enzyme. A strandof Dicer-generated small RNA duplex (called the guide strand)is then selected by a thermodynamic mechanism to associate withArgonaute (AGO) protein. This AGO–small RNA complex functionsto cleave mRNA, repress translation or modify chromatin structurein a sequence-specific manner. Although a model plant, Arabidopsisthaliana, contains 10 AGO genes, their roles and molecular mechanismsremain obscure. In this study, we analyzed the roles of ArabidopsisAGO2 and AGO5. Interestingly, the 5' nucleotide of small RNAsthat associated with AGO2 was mainly adenine (85.7%) and thatwith AGO5 was mainly cytosine (83.5%). Small RNAs that wereabundantly cloned from the AGO2 immunoprecipitation fraction(miR163-LL, which is derived from the Lower Left of mature miR163in pre-miR163, and miR390) and from the AGO5 immunoprecipitationfraction (miR163-UL, which is derived from the Upper Left ofmature miR163 in pre-miR163, and miR390^*) are derived from thesingle small RNA duplexes, miR163-LL/miR163-UL and miR390/miR390^*.Each strand of the miR163-LL/miR163-UL duplex is selectivelysorted to associate with AGO2 or AGO5 in a 5' nucleotide-dependentmanner rather than in a thermodynamic stability-dependent manner.Furthermore, we showed that both AGO2 and AGO5 have the abilityto bind cucumber mosaic virus-derived small RNAs. These resultsclearly indicate that the mechanism selecting the guide strandis different among AGO proteins and that multiple AGO genesare involved in anti-virus defense in plants.     

Comparison of plasma viremia and antibody responses in macaques inoculated with envelope variants of single-cycle simian immunodeficiency virus differing in infectivity and cellular tropism

Molecular differences in the envelope glycoproteins of human immunodeficiency virus type 1 and simian immunodeficiency virus (SIV) determine virus infectivity and cellular tropism. To examine how these properties contribute to productive infection in vivo, rhesus macaques were inoculated with strains of single-cycle SIV (scSIV) engineered to express three different envelope glycoproteins with full-length (TM_open) or truncated (TM_stop) cytoplasmic tails. The 239 envelope uses CCR5 for infection of memory CD4⁺ T cells, the 316 envelope also uses CCR5 but has enhanced infectivity for primary macrophages, and the 155T3 envelope uses CXCR4 for infection of both naive and memory CD4⁺ T cells. Separate groups of six rhesus macaques were inoculated intravenously with mixtures of TM_open and TM_stop scSIV_mac239, scSIV_mac316, and scSIV_mac155T3. A multiplex real-time PCR assay specific for unique sequence tags engineered into each virus was then used to measure viral loads for each strain independently. Viral loads in plasma peaked on day 4 for each strain and were resolved below the threshold of detection within 4 to 10 weeks. Truncation of the envelope cytoplasmic tail significantly increased the peak of viremia for all three envelope variants and the titer of SIV-specific antibody responses. Although peak viremias were similar for both R5- and X4-tropic viruses, clearance of scSIV_mac155T3 TM_stop was significantly delayed relative to the other strains, possibly reflecting the infection of a CXCR4⁺ cell population that is less susceptible to the cytopathic effects of virus infection. These studies reveal differences in the peaks and durations of a single round of productive infection that reflect envelope-specific differences in infectivity, chemokine receptor specificity, and cellular tropism.     

Rice alpha-mannosidase digesting the high mannose glycopeptide of glutelin

α -Mannosidase (EC 3.2.1.24) from rice dry seeds was purified to homogeneity. Optimum pH and K_m for pNP- α -Man hydrolysis were pH 4.3–4.5 and 1.04 m M , respectively. The enzyme digested mannobioses such as Man α -1,2Man, Man α -1,6Man, Man α -1,3Man but Man α -1,4Man. Zn²⁺ ion was required for the activity, whereas EDTA and swainsonine inhibited the activity by 80 and 96%, respectively. The rice storage protein, glutelin was prepared and its basic subunits were shown to have high mannose-type sugar chains by two-dimensional mapping using NH₂-P and C₁₈ silica columns. They were Man₉GlcNAc₂, Man₈GlcNAc₂, Man₇GlcNAc₂, Man₆GlcNAc₂ and Man₅GlcNAc₂. All these oligosaccharides were digested by the purified α -mannosidase, and Man-GlcNAc₂ and mannose were formed. Glycopeptides, having these high mannose-type sugar chains, could also be digested by the α -mannosidase. Subunits were prepared from glutelin basic subunit and the richest subunit among them, subunit 2 (isoform 2), was digested by the α -mannosidase. Isoform 2 was digested by V8 protease only partially and slowly. However, isoform 2, pre-treated with the α -mannosidase, was rapidly and completely digested by V8 protease.