Trajectory of criterion symptoms of major depression under newly started antidepressant treatment: sleep disturbances and anergia linger on while suicidal ideas and psychomotor symptoms disappear early.

OBJECTIVE: In modern psychiatry, depression is diagnosed with the diagnostic criteria; however, the trajectory of each of the criterion symptoms is unknown. This study aims to examine this. METHODS: We made repeated assessments of the nine diagnostic criterion symptoms with the Patient Health Questionnaire-9 (PHQ-9) among 2011 participants of a 25-week pragmatic randomised controlled trial of sertraline and/or mirtazapine for hitherto untreated major depressive episodes. The changes from baseline were estimated with the mixed-effects model with repeated measures. The time to disappearance of each symptom was modeled using the Kaplan-Meier survival analysis. RESULTS: The total score on PHQ-9 was 18.5 (SD = 3.9, n = 2011) at baseline, which decreased to 15.3 (5.2, n = 2011) at week 1, to 11.5 (5.9, n = 1953) at week 3, to 7.8 (6.0, n = 1927) at week 9, and to 6.0 (5.9, n = 1910) at week 25. Suicidal ideas, psychomotor symptoms decreased rapidly, while anergia and sleep disturbance also decreased but only slowly. The survival analyses confirmed the primary analyses. CONCLUSIONS: Upon initiation of antidepressant treatment, patients with newly treated major depressive episodes can expect their suicidal ideas and psychomotor symptoms to disappear first but sleep disturbances and anergia to linger on.

Time-dependent distinct roles of Toll-like receptor 4 in a house dust mite-induced asthma mouse model.

House dust mites (HDMs) are a common source of allergens that trigger both allergen-specific and innate immune responses in humans. Here, we examined the effect of allergen concentration and the involvement of Toll-like receptor 4 (TLR4) in the process of sensitization to house dust mite allergens in an HDM extract-induced asthma mouse model.　Intranasal administration of HDM extract induced an immunoglobulin E response and eosinophilic inflammation in a dose-dependent manner from 2.5 to 30 µg/dose. In TLR4-knockout mice, the infiltration of eosinophils and neutrophils into the lung was decreased compared with that in wild-type mice in the early phase of inflammation (total of three doses). However, in the late phase of inflammation (total of seven doses), eosinophil infiltration was significantly greater in TLR4-knockout mice than in wild-type mice. This suggests that the roles of TLR4 signaling are different between the early phase and the later phase of HDM allergen-induced inflammation. Thus, innate immune response through TLR4 regulated the response to HDM allergens, and the regulation was altered during the phase of inflammation.

Body odour aldehyde reduction by acetic acid bacterial extract including enzymes: alcohol dehydrogenase and aldehyde dehydrogenase.

OBJECTIVE: The major causes of unpleasant human body odour are aldehydes produced by axillary-resident bacteria. There are many methods of body odour prevention; however, they all carry risks of destroying indigenous dermal bacteria that are necessary for the maintenance of the normal physical function of the skin. Furthermore, some methods cannot directly reduce the concentrations of substances that cause body odour. Therefore, a novel method of reducing body odour more safely and effectively is required. We focused on acetic acid bacterial enzymes, which can convert aldehydes into carboxylic acids, and investigated their effect on aldehydes and body odour. METHODS: Subjects with strong body odour were recruited using screening questionnaires. Acetic acid bacterial extract including enzymes was applied to subjects' skin, and their effects were evaluated by trained panellists and by quantitative aldehyde analysis using thermal detector gas chromatography/mass spectrometry. RESULTS: Acetic acid bacterial extract including enzymes decreased the ratio of dilution to threshold and the concentration of body odour-producing aldehydes dropped by up to 98.7%. CONCLUSION: These results indicate that simply applying acetic acid bacterial enzymes on the skin can reduce the concentration of aldehydes that cause unpleasant body odour by directly converting them into carboxylic acids. Therefore, acetic acid bacterial enzymes can potentially be developed into new products that do not destroy indigenous bacteria and yet can effectively reduce unpleasant body odour.

Tumour-infiltrating CD8 to FOXP3 lymphocyte ratio in predicting treatment responses to neoadjuvant chemotherapy of aggressive breast cancer.

BACKGROUND: Tumour-infiltrating lymphocytes (TILs) can be used to monitor the immune response, and are important in predicting treatment responses and outcomes for various types of cancer. Recently, specific TIL subsets have been reported to be clinically useful in predicting treatment responses. The CD8+/FOXP3+ TIL ratio (CFR) may be a more sensitive indicator for monitoring immune function. This study investigated the clinical significance and value of CFR as a biomarker to predict treatment responses to neoadjuvant chemotherapy for breast cancer. METHODS: Patients with resectable early-stage breast cancer treated with neoadjuvant chemotherapy at Osaka City University Hospital, Japan, between 2007 and 2013 were included. Oestrogen receptor, progesterone receptor, human epidermal growth factor receptor (HER) 2, Ki-67, CD8 and FOXP3 status were assessed by immunohistochemistry, and correlated with pathological complete response (pCR). RESULTS: A total of 177 patients were included, of whom 90 had a high CFR and 87 a low CFR. Triple-negative breast cancer (TNBC) was more common in the high-CFR group than in the low-CFR group (46 versus 23 per cent; P = 0·002), as was HER2-enriched breast cancer (HER2BC) (27 versus 14 per cent; P = 0·033). Among these patients, the pCR rate was significantly higher in the high-CFR group than in the low-CFR group (TNBC: P = 0·022; HER2BC: P < 0·001). In multivariable analysis high-CFR status was an independent predictor of a favourable prognosis: hazard ratio 0·24 (95 per cent c.i. 0·05 to 0·72; P = 0·015) for TNBC and 0·10 (0·10 to 0·90; P = 0·041) for HER2BC. CONCLUSION: The CFR may be a useful biomarker to predict treatment response to neoadjuvant therapy in aggressive breast cancer subtypes, such as TNBC and HER2BC.

Measurement of membrane hydraulic conductivity of bovine carotid artery endothelial cells using a perfusion microscope.

The osmotic properties of bovine carotid artery endothelial cells (BCAECs) associated with cryobiology were investigated using a perfusion microscope. These properties include the hydraulic conductivity (Lp) and its activation energy (ELp). The response of isolated cells was observed when the extracellular concentration increased from 0.15 M to 0.5 M NaCl at three different temperatures. The transient volumes of the cell were calculated from the measurements of the projected areas with an assumption of a spherical cell. The hydraulic conductivity (Lp) and the osmotically inactive volume (Vb) of BCAECs were simultaneously determined using nonlinear regression to fit the change of cell volume estimated by water transport equations to measured cell volumes. The Lp values were 0.26 +/- 0.08, 0.12 +/- 0.02, and 0.06 +/- 0.02 m/atm/min (mean +/- SD) at 23, 11 and 4 degree C, respectively, yielding the activation energy of Lp of 47.6 kJ/mol according to the Arrhenius relationship.

Detection of patients with cancer by monoclonal antibody directed to lactoneotetraosylceramide (paragloboside).

A hybridoma producing monoclonal antibody (H11) directed to lactoneotetraosylceramide (paragloboside) has been established from spleen cells of a mouse immunized with paragloboside. The monoclonal antibody H11 (immunoglobulin M type) was selected from five clones showing different reactivities with paragloboside. The monoclonal antibody was highly specific to paragloboside and lacked reactivity with other glycolipids including glucosylceramide, lactosylceramide, globotriaosylceramide, globotetraosylceramide, gangliotriaosylceramide, gangliotetraosylceramide, and GalNAc beta 1-4[NeuAc alpha 2-3]Gal beta 1-4Glc beta 1-1Cer. However, the monoclonal antibody (H11) was found to bind to lactosamine-containing glycolipids at their terminals, such as i- and I-type glycolipids as well as paragloboside. A two-step sandwich radioimmunoassay method for paragloboside antigen in serum was established by using the monoclonal antibody. The mean paragloboside antigen concentration in the sera from 20 normal individuals was 25.3 ng/ml. If the cutoff value was set at 80.9 ng/ml [25.3 + 2 x 27.8 (SD)], only 1 of 20 healthy controls had an elevated paragloboside value in the serum, whereas sera from 9 of 12 (75.0%) hepatoma, 4 of 10 (40%) pancreatic cancer, 16 of 40 (40.0%) stomach cancer, and 6 of 10 (60%) lung cancer patients had elevated paragloboside values. Sera from 3 of 8 hepatitis patients and 7 of 10 liver cirrhosis patients were estimated to be positive but sera from 16 patients with benign disease had paragloboside levels lower than the cutoff value. A larger amount of the antigen was found in liver metastases from colorectal carcinoma compared to the normal counterpart. The antigen was also detected in the medium of various human cancer cells and meconium. However, the antigen in the sera, medium, meconium, and cancer tissue seemed to be associated with glycoprotein or lipoprotein, because most of the antigen activity was eluted in the void volume fraction on high-performance liquid chromatography with a gel filtration column.

Accumulation of gangliosides with N-acetylneuraminosyl(alpha 2-6)lactosamine structure in primary human hepatoma.

Gangliosides of hepatomas have been analyzed by using a monoclonal antibody directed to N-acetylneuraminosyl(alpha 2-6)lactoneotetraosylceramide (sialyl(alpha 2-6)paragloboside), which was prepared by injecting the monosialoganglioside fraction of human meconium into BALB/c mice. The monoclonal antibody, named MSG-15, was found to bind sialyl(alpha 2-6)paragloboside, but it failed to react with other gangliosides, including N-acetylneuraminosyl(alpha 2-3)lactoneotetraosylceramide (sialyl (alpha 2-3)paragloboside) and "Ii"-type gangliosides. MSG-15 was found to recognize NeuAc alpha 2-6Gal beta structure of the ganglioside. Gangliosides obtained from human hepatomas were analyzed by immunostaining on high-performance thin-layer chromatography plates using the monoclonal antibody MSG-15. All primary hepatoma samples used in this study (nine samples) were found to contain sialyl(alpha 2-6)paragloboside, which accounted for 13-31% of the monosialoganglioside fractions in the hepatomas. Furthermore, MSG-15 recognized several monosialogangliosides in addition to sialyl(alpha 2-6)paragloboside. These gangliosides apparently also contain a terminal NeuAc alpha 2-6Gal beta structure. Other ganglioside fractions obtained from hepatoma and meconium were immunostained on thin layer chromatography plates with MSG-15. Additionally, another monoclonal antibody (H-11), which recognizes terminal lactosamine structure, was used to immunostain these fractions after sialidase treatment. Bands stained with both monoclonal antibodies showed similar mobilities to each other in the di- and trisialoganglioside fractions as well as monosialoganglioside fraction. In control liver, GM3 ganglioside accounted for 92% of monosialoganglioside fraction, and sialyl(alpha 2-6)paragloboside accounted for less than 1% of the fraction. Immunohistochemical study by using MSG-15 in tissue sections from hepatocellular carcinoma and normal liver tissues demonstrated that only hepatocellular carcinoma cells gave a positive reaction. These results suggest that the biosynthetic pathway of gangliosides containing NeuAc alpha 2-6Gal beta 1-4GlcNAc beta structure is activated in hepatoma cells.

Effect of vitamin K2 on three-dimensional trabecular microarchitecture in ovariectomized rats.

Menatetrenone, a vitamin K2 with four isoprene units, has been reported to improve osteoporotic bone loss. The purpose of this investigation was to clarify the effect of menatetrenone on the three-dimensional (3D) trabecular microarchitecture in ovariectomized (OVX) rats by using microcomputed tomography (MCT). Forty-two 13-week-old female rats were used and divided into four groups: the OVX (OVX + MK-4) group treated with menatetrenone, the (OVX untreated) group, the sham-operated (Sham + MK-4) group treated with menatetrenone, and the sham-operated group not treated with menatetrenone (Sham untreated) group. OVX rats were fed a calcium-deficient diet. Menatetrenone treatment was begun just after the ovariectomy, and the mean menatetrenone oral intake over the 8-week period was adjusted to 30 mg/kg BW per day. The proximal metaphyseal region of the right tibia was evaluated by dual X-ray absorptiometry (DXA) and MCT. A parametric analysis of the reconstructed trabecular volume was carried out using bone volume fractions, the fractal dimension calculated by the 3D box-counting method, and the connectivity density as determined by topological analysis. Menatetrenone significantly increased the trabecular bone volume, fractal dimension, and connectivity in the OVX + MK-4 group compared with the OVX-untreated group (p < 0.01). Our results suggest that an 8-week administration of menatetrenone protects against the loss of trabecular bone volume and its connectivity when treatment is begun just after the ovariectomy. Despite this apparent protection, it remains unknown whether it is possible to reestablish trabecular connectivity if therapeutic intervention occurs after the trabecular connectivity has been lost.

Mechanical strain effect on bone-resorbing activity and messenger RNA expressions of marker enzymes in isolated osteoclast culture.

Adaptive modeling and remodeling are controlled by the activities of osteoblasts and osteoclasts, which are capable of sensing their mechanical environments and regulating deposition or resorption of bone matrix. The effects of mechanical stimuli on isolated osteoclasts have been scarcely examined because it has proven to be difficult to prepare a number of pure osteoclasts and to cultivate them on mineralized substratum during mechanical stimulation. Recently, we developed an apparatus for applying mechanical stretching to the ivory slice/plastic plate component on which cells could be cultured. The loading frequency, strain rate, and generated strain over an ivory surface could be controlled by a personal computer. Using this apparatus, we examined the role of mechanical stretching on the bone-resorbing activity of the osteoclasts. Mature and highly enriched osteoclasts were cultured for 2, 12, and 24 h on the ivory/plate component while being subjected to intermittent tensile strain. The stretched osteoclasts showed enhanced messenger RNA (mRNA) expression levels of osteoclast marker enzymes, tartrate-resistant acid phosphatase (TRAP), and cathepsin K and increases of resorbed-pit formation, suggesting that the mechanical stretching up-regulated the bone-resorbing activity of the osteoclasts. A stretch-activated cation (SA-cat) channel blocker significantly inhibited the increases of the mRNA level and pit formation after 24 h of stretching. This study suggested the possibility that the mature osteoclasts responded to mechanical stretching through a mechanism involving a SA-cat channel in the absence of mesenchymal cells and, as a result, up-regulated their bone-resorbing activity.

Measurement of plasma renin activity by a simple solid phase radioimmunoassay.

A solid phase RIA in which an antibody adsorbed onto polystyrene balls was developed to determine PRA. Complete inhibition of converting enzyme and angiotensinase during enzymatic reaction was achieved by phenyl methyl sulfonyl fluoride and EDTA combination. Pepstatin A was found to be an effective agent to block angiotensin I generation during the RIA, and the sample can be directly incubated at room temperature for RIA without any special treatment to inhibit renin activity. The intra- and interassay coefficients of variation (CV) were 5.5-6.9% and 3.7-8.2%, respectively. The recovery was 91.9-117% of added angiotensin I. The assay is sufficiently sensitive and reliable for routine use and correlates acceptably (r = 0.996) with an established RIA. The antibody-adsorbed balls were compared to the soluble antibody with respect to thermodynamic parameters. It was found that the apparent equilibrium constant of the antibody-adsorbed ball was reduced to approximately 1/2 sec of soluble antibody, which was predominately due to the decrease in unitary entropy change by adsorption.

Comparative study of dopamine metabolism with local cerebral glucose utilization in rat brain following the administration of haloperidol decanoate.

The effects of haloperidol decanoate on dopamine (DA) metabolism in discrete regions of rat brain were investigated and compared with changes in local cerebral glucose utilization (LCGU). The concentration of DA and its metabolite, homovanillic acid, and the alpha-methyl-p-tyrosine (alpha-MT)-induced decline of DA were measured in 6 brain regions by a high-performance liquid chromatographic assay. LCGU in 26 brain regions were examined by [14C]2-deoxy-D-glucose autoradiography. At 24-hr after intramuscular injection of haloperidol decanoate (60 mg eq/kg to haloperidol), the concentration of homovanillic acid in the prefrontal cortex, caudate-putamen, accumbens nucleus, lateral amygdala, and medial thalamus showed significant increase compared with control values. On day 21, the increase in these regions was significantly attenuated with no significant difference from the controls. Furthermore, chronic haloperidol rats showed alpha-MT-induced decline of DA to a similar extent in the control rats. LCGU on day 21 showed significant decrease in the parietal cortex, and a tendency toward decrease in the prefrontal cortex, lateral amygdala and medial thalamus compared with the controls. There was no significant change in LCGU in the caudate-putamen or accumbens nucleus. Chronic haloperidol would thus appear to affect energy metabolism mainly in the cortico-thalamo-limbic circuits, and this may not correspond well to presynaptic DA metabolism.

Preparation of various Tc-99m dimercaptosuccinate complexes and their evaluation as radiotracers.

The organ distributions of four different Tc-99m dimercaptosuccinate complexes ("Tc-DMS", Complexes 1 to 4) were determined using mice and were evaluated as renal imaging agents. The highest kidney uptake was observed with Complex 2:21.7% of dose, 3 hr after injection. The biologic distributions and gel chromatographic analyses using carrier Tc-99 and Sn-113 indicate that there is little possibility of mixed metal complexes of the type Tc-Sn-DMS: rather they contain only Tc + DMS. The labeling procedure for Tc-99m DMS as a renal agent proceeds in two steps: a rapid formation of Complex 1, and a slower, rate-determining step from Complex 1 to Complex 2. A reproducible lyophilized kit has been prepared. The yield of complex 2 greatly depends on the reconstitution volume of 99mTcO4-; yield averages 89% using 2 ml of 99mTcO4-eluate.

New preparation method for 99mTc-phytate.

A new method is described in which Pt-Sn or Sn-Sn electrodes are used to activate 99mTcO4-. The 99mTc is incorporated into phytic acid by stannous ion released from a tin anode by the corrosive reaction. The most suitable pH for labeling phytic acid by this method was below 5 and the 99mTc-phytate formed could be precipitatedd with Ca+2 at a pH above 3.5. Though 99mTc-phytate is soluble in aqueous solution, it forms an insoluble species with Ca+2 in vivo and is trapped in the reticuloendothelial system. More than 93% of the 99mTc-phytate localized in the liver of mice; here its biologic half-time is about 112 hr. The 99mTc-phytate prepared by this method promises to be useful as a liver-scanning agent. Since our method does not require an applied electric potential, it appears to be one of the most convenient methods for labeling compounds with 99mTc.

Distribution of clomipramine in various brain regions of rats under steady-state serum concentrations.

The clomipramine (CMP) distribution in 12 regions of rat brain was investigated 2 and 7 days after subcutaneous minipump implantation. The results of serial blood samplings during 7 days indicated that steady-state CMP serum levels were achieved from experimental day 2. The regional CMP distributions in the brains were uneven on both the 2nd and 7th experimental days. On both test days the CMP concentrations were higher in the cerebral cortex and hippocampus than those in the other regions, and those in the cerebellum and bulbus olfactorius and septum were lower. When the serial fluctuations of the CMP levels were examined in 12 brain regions, there were significant differences between days 2 and 7 in the following four regions: the anterior basal ganglia, hypothalamus, bulbus olfactorius + septum and amygdala.

Structural evaluation of glucose analogues on feeding elicitation in rat.

The effects of 12-mumol doses of the glucose analogues glucosamine, 2-fluoroglucose, 2-chloroglucose, 2-deoxyglucose (which were modified at carbon 2 of the glucopyranose ring), 1-aminoglucose and 1-deoxyglucose (modified at carbon 1), on feeding behavior and plasma glucose, insulin, and glycerol were examined after infusion into the rat third cerebroventricle. Plasma glucose and glycerol levels were elevated by glucosamine or 1-aminoglucose. Plasma insulin levels were unchanged by these analogues. Feeding was induced in 62% to 87% of the rats tested after infusion of glucosamine, 2-fluoroglucose, 2-chloroglucose, 2-deoxyglucose, 1-aminoglucose, or 1-deoxyglucose (mean meal size in responding rats, 43.9, 25.8, 22.7, 16.0, 42.3, and 3.8 pellets, respectively). The order of potency to induce feeding was amino, halogen, and hydrogen groups. These data reinforced the concept that the potency of glucose analogues to induce feeding depends on substituents at carbon 1 and carbon 2 of the glucopyranose ring.

Deformation Theory of Dynamic Link Matching

Dynamic link matching is a self&hyphenorganizing topographic mapping between a template image and a data image. The mapping tends to be continuous, linking two points sharing similar local features, which, as a result, can lead to its deformation to some degree. In analyzing such deformation mathematically, we reduced the model equation to a phase equation, which enabled us to clarify the principles of the deformation process and the relationship between high&hyphendimensional models and low&hyphendimensional ones. We also elucidated the characteristics of the model in the context of the standard regularization theory.

Site of origin of projections from the thalamus to dorsal versus ventral aspects of the premotor cortex of monkeys.

Retrograde tracers were injected into the forelimb regions of three cortical motor areas: (1) a dorsal aspect of the premotor cortex (PMd) immediately lateral to the superior precentral sulcus; (2) a ventral aspect of the premotor cortex (PMv) immediately caudal to the genu of the arcuate sulcus and lateral to the arcuate spur; and (3) the primary motor cortex (MI). Before tracer injection, single-unit recordings were made to select injection sites in the forelimb regions where neurons with set- and/or movement-related activity before forelimb movements were densely located. Following the PMd injections, labeled cells were found mainly in rostral portion of VLc and VLo. Cells projecting to PMv were found mainly in X and VPLo. Projection cells to the MI were found in VPLo, VLc, and VLo. Locations of neurons projecting to different motor areas were not overlapped in the thalamic nuclei. Combining available reports, the results suggest that major inputs to PMd come from globus pallidus and that, in contrast, cerebellum is a main source to the PMv. The differential inputs to PMd and PMv may contribute to their functional specialization.

Corticocortical inputs to the dorsal and ventral aspects of the premotor cortex of macaque monkeys.

Recent cytoarchitectonic, histochemical and physiological studies have shown that the lateral part of area 6 (the premotor cortex) of macaque monkeys can be divided into at least two subregions, each of which is considered to play an important role in motor control. One lies in the dorsal aspect of the premotor cortex (PMd) medial to the spur of the arcuate sulcus, and the other in the ventral aspect of the premotor cortex (PMv) lateral to it. Since there is little information on the corticocortical inputs to the PMd, wheat-germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) was injected into both the PMd and PMv to study corticocortical inputs to these two regions, and the distribution of retrogradely labeled cells was compared. When WGA-HRP was injected into the region immediately lateral to the superior precentral sulcus within the PMd, retrogradely labeled neurons were found in area 6 lying in the mesial wall possibly corresponding to the supplementary motor area (SMA), areas 24 and 23 of the cingulate cortex, rostral region of area 4, and area 5 (area PEa). In contrast, when WGA-HRP was injected into the PMv immediately caudal to the arcuate sulcus and lateral to the spur of the arcuate sulcus, the labeled cells were found in area 7 (areas POa, PF, PFG), area 5 (area PEa), area PFop (secondary somatosensory area), SMA, the cingulate cortex (areas 24), caudal region of area 4 in the rostral bank of the central sulcus, and area 3a. It appears that the differences in the corticocortical inputs contribute to specialization of the PMd and PMv for their differential roles in motor control.

Information processing for motor control in primate premotor cortex.

Recent neurophysiological as well as neuropsychological studies provided evidences on how various informations are processed to generate motor programs in the central nervous system. In this article, functional specializations of two distinct cortical motor areas, the dorsal and ventral aspects of the premotor cortex (PMd and PMv, respectively) of macaque monkeys, are focused to review this issue. Three major conclusions emerged from recent neurophysiological studies. First, each of movement parameters such as amplitude and direction is distinctively programmed in PMd by serial integration, rather than by parallel distributed processing. Second, in performance of conditional motor behavior, conditionally presented sensory signals are processed for motor preparation and execution of an intended act in PMd, but not in PMv. Third, PMv may be specialized for motor execution under visual guidance.

Responses of pyramidal tract neurons in the postcentral cortex to tactile inputs.

Pyramidal tract neurons were recorded from postcentral cortex of awake monkeys and their responses to step indentation and vibratory stimulus were studied. The majority of them exhibited slowly adapting response to the indentation stimulus but failed to show phase-locked response to 50-200 Hz vibrations. The response properties appeared to be in contrast to those of non-pyramidal tract neurons whose responses were largely quickly adapting.