Oxygen-evolving photosystem II structures during S1-S2-S3 transitions.

Photosystem II (PSII) catalyses the oxidation of water through a four-step cycle of Si states (i = 0-4) at the Mn4CaO5 cluster1-3, during which an extra oxygen (O6) is incorporated at the S3 state to form a possible dioxygen4-7. Structural changes of the metal cluster and its environment during the S-state transitions have been studied on the microsecond timescale. Here we use pump-probe serial femtosecond crystallography to reveal the structural dynamics of PSII from nanoseconds to milliseconds after illumination with one flash (1F) or two flashes (2F). YZ, a tyrosine residue that connects the reaction centre P680 and the Mn4CaO5 cluster, showed structural changes on a nanosecond timescale, as did its surrounding amino acid residues and water molecules, reflecting the fast transfer of electrons and protons after flash illumination. Notably, one water molecule emerged in the vicinity of Glu189 of the D1 subunit of PSII (D1-E189), and was bound to the Ca2+ ion on a sub-microsecond timescale after 2F illumination. This water molecule disappeared later with the concomitant increase of O6, suggesting that it is the origin of O6. We also observed concerted movements of water molecules in the O1, O4 and Cl-1 channels and their surrounding amino acid residues to complete the sequence of electron transfer, proton release and substrate water delivery. These results provide crucial insights into the structural dynamics of PSII during S-state transitions as well as O-O bond formation.

Evidence of unconventional superconductivity on the surface of the nodal semimetal CaAg1-xPdxP.

Surface states of topological materials provide extreme electronic states for unconventional superconducting states. CaAg1-xPdxP is an ideal candidate for a nodal-line Dirac semimetal with drumhead surface states and no additional bulk bands. Here, we report that CaAg1-xPdxP has surface states that exhibit unconventional superconductivity (SC) around 1.5 K. Extremely sharp magnetoresistance, tuned by surface-sensitive gating, determines the surface origin of the ultrahigh-mobility "electrons." The Pd-doping elevates the Fermi level towards the surface states, and as a result, the critical temperature (Tc) is increased up to 1.7 K from 1.2 K for undoped CaAgP. Furthermore, a soft point-contact study at the surface of Pd-doped CaAgP proved the emergence of unconventional SC on the surface. We observed the bell-shaped conductance spectra, a hallmark of the unconventional SC. Ultrahigh mobility carriers derived from the surface flat bands generate a new class of unconventional SC.

Sensory processing in children and adolescents shortly after the onset of anorexia nervosa: a pilot study.

BACKGROUND: Alterations in sensory processing, such as vision, taste, and interoceptive sensation, have been reported in adult anorexia nervosa (AN). Whether these symptoms are traits, states, or "scars" due to chronic starvation has not been fully established. Based on the hypothesis that alterations in sensory processing also occur in adolescent AN in the early stages of the disease, the present study was conducted using both self-administered and parent-administered sensory processing questionnaires. METHODS: Children and adolescents with anorexia nervosa treated at a single tertiary eating disorder treatment center in Japan (AN group) and female junior high school students attending a public junior high school in Saitama Prefecture, Japan (healthy control group: HC group) were included in the study. The Sensory Profile (SP) and Adult/Adolescent Sensory Profile (AASP) were administered to the participants and their caregivers. In addition, we collected demographic data and administered the Children's Eating Attitude Test and Autism-Spectrum Quotient Children's version. RESULTS: Seventeen children and adolescents were enrolled in the AN group, and 63 were enrolled in the HC group. There was no statistically significant difference between the AN and HC groups in the quadrant scores of the AASP. In the SP, the Sensory Avoiding score and the Emotional/Social response score were higher in the AN group than in the HC group. CONCLUSION: From the parents' point of view, the patient avoids unexpected sensory stimuli, but the patients are unaware of their own avoiding behavior in the early stages of the disease. The results suggest that sensory sensitivity in AN may be a "scar" symptom due to chronic starvation and a state symptom. Longitudinal studies from shortly after the onset with larger sample sizes are needed to gain insight into the dynamic relation between sensory processing and eating disorder pathology.

Excited-state intermediates in a designer protein encoding a phototrigger caught by an X-ray free-electron laser.

One of the primary objectives in chemistry research is to observe atomic motions during reactions in real time. Although X-ray free-electron lasers (XFELs) have facilitated the capture of reaction intermediates using time-resolved serial femtosecond crystallography (TR-SFX), only a few natural photoactive proteins have been investigated using this method, mostly due to the lack of suitable phototriggers. Here we report the genetic encoding of a xanthone amino acid (FXO), as an efficient phototrigger, into a rationally designed human liver fatty-acid binding protein mutant (termed XOM), which undergoes photo-induced C-H bond transformation with high selectivity and quantum efficiency. We solved the structures of XOM before and 10-300 ns after flash illumination, at 1.55-1.70 Å resolutions, and captured the elusive excited-state intermediates responsible for precise C-H bond activation. We expect that most redox enzymes can now be investigated by TR-SFX, using our method, to reveal reaction intermediates key for their efficiency and selectivity.

Adolescent Health Promotion Interventions Using Well-Care Visits and a Smartphone Cognitive Behavioral Therapy App: Randomized Controlled Trial.

BACKGROUND: Adolescent health promotion is important in preventing risk behaviors and improving mental health. Health promotion during adolescence has been shown to contribute to the prevention of late onset of the mental health disease. However, scalable interventions have not been established yet. OBJECTIVE: This study was designed to test the efficacy of two adolescent health promotion interventions: a well-care visit (WCV) with a risk assessment interview and counseling and self-monitoring with a smartphone cognitive behavioral therapy (CBT) app. Our hypothesis was that participants who had received both WCV and the CBT app would have better outcomes than those who had received only WCV or those who had not received any intervention. We conducted a prospective multi-institutional randomized controlled trial. METHODS: Participants were 217 adolescents aged 13-18 years. They were randomly divided into two intervention groups (WCV group and WCV with CBT app group) and a nonintervention group. WCV comprised a standardized physical examination along with a structured interview and counseling for youth risk assessment, which was designed with reference to the Guideline for Health Supervision of Adolescents of Bright Futures. A smartphone-based CBT program was developed based on the CBT approach. The CBT app comprised a 1-week psychoeducation component and a 1-week self-monitoring component. During the CBT program, participants created several self-monitoring sheets based on the CBT model with five window panels: event, thoughts, feelings, body response, and actions. The primary outcome was the change in scores for depressive symptoms. Secondary outcomes included changes in scores for self-esteem, quality of life, self-monitoring, and an adolescent health promotion scale. These outcomes were evaluated at baseline and at 1, 2, and 4 months after baseline. The exploratory outcome was the presence of suicidal ideation during the observation period. Intervention effects were estimated using mixed effect models. RESULTS: In total, 94% (204/217) of the participants completed the 4-month evaluation. Both intervention groups showed a significant effect in the form of reduced scores for depressive symptoms at 1 month in high school students; however, these effects were not observed at 2 and 4 months. The intervention effect was significantly more predominant in those scoring above cutoff for depressive symptoms. There was significantly less suicidal ideation in the intervention groups. As for secondary outcomes, there was significant increase in health promotion scale scores at the 4-month follow-up among junior high school students in the WCV group. Moreover, the CBT app was significantly effective in terms of obtaining self-monitoring skills and reducing depressive symptoms. CONCLUSIONS: Although adolescent health promotion interventions may have short-term benefits, the frequency of WCV and further revision of the CBT app should be considered to evaluate long-term effectiveness. TRIAL REGISTRATION: University Hospital Medical Information Network Clinical Trials Registry UMIN 000036343; https://center6.umin.ac.jp/cgi-open-bin/ctr/ctr_view.cgi?recptno=R000041246.

Cerebellar tonsillar herniation in sudden death of an adolescent anorexia nervosa patient: a case report.

BACKGROUND: Anorexia nervosa not only results in severe malnutrition but also carries a high risk of sudden death. Although fatal arrhythmias are the most common cause of sudden death, it is often unclear what exactly causes them. To the best of our knowledge, there have been no reports of cerebellar tonsillar herniations in patients with anorexia nervosa. CASE PRESENTATION: A 17-year-old girl with anorexia nervosa and autism spectrum disorder was admitted to the pediatric ward for extreme weight loss. Since she refused to take oral nutrition or tube feeding, we started continuous intravenous fluids. Eight hours after admission, she suddenly went into cardiopulmonary arrest and died despite resuscitation. A postmortem autopsy revealed the unexpected findings of generalized severe cerebral edema and cerebellar tonsillar herniation. CONCLUSION: Intracranial hypertension may need to be considered when the condition of patients with anorexia nervosa suddenly worsens during refeeding periods. Postmortem autopsy and autopsy imaging are recommended to determine the exact cause of sudden death. LEVEL OF EVIDENCE: Level IV: Evidence obtained from multiple time series analysis such as case studies. (NB: Dramatic results in uncontrolled trials might also be regarded as this type of evidence).

Pyosalpinx due to Cronobacter sakazakii in an elderly woman.

BACKGROUND: Cronobacter sakazakii (C. sakazakii) is a bacterium known to cause severe neonatal infections in premature infants with the consumption of contaminated powdered milk formula. Adult infections are rare, and there have been no reports of pyosalpinx due to C. sakazakii to date. CASE PRESENTATION: We report a case of left pyosalpinx due to C. sakazakii in a sexually inactive postmenopausal woman. A 70-year-old woman presented to our hospital with left lower abdominal pain and fever. Abdominal computed tomography disclosed a cystic mass continuous with the left edge of the uterus. Urgent laparotomy revealed a ruptured left pyosalpinx with pus-like content. Left salpingo-oophorectomy, resection of the right tube, and washing of the abdominal cavity with saline were performed. Pathological examination of the left adnexa showed tubal tissue with acute inflammation and inflammatory exudate, which were compatible with pyosalpinx, and pus culture yielded C. sakazakii. CONCLUSIONS: This is the first case report of pyosalpinx due to C. sakazakii. Cronobacter sakazakii infections in adult women might occur in the elderly, whose immunity has weakened. Further accumulation of cases of C. sakazakii infection is needed to clarify the etiology and behavior of C. sakazakii in adults.

Bone suppression for chest X-ray image using a convolutional neural filter.

Chest X-rays are used for mass screening for the early detection of lung cancer. However, lung nodules are often overlooked because of bones overlapping the lung fields. Bone suppression techniques based on artificial intelligence have been developed to solve this problem. However, bone suppression accuracy needs improvement. In this study, we propose a convolutional neural filter (CNF) for bone suppression based on a convolutional neural network which is frequently used in the medical field and has excellent performance in image processing. CNF outputs a value for the bone component of the target pixel by inputting pixel values in the neighborhood of the target pixel. By processing all positions in the input image, a bone-extracted image is generated. Finally, bone-suppressed image is obtained by subtracting the bone-extracted image from the original chest X-ray image. Bone suppression was most accurate when using CNF with six convolutional layers, yielding bone suppression of 89.2%. In addition, abnormalities, if present, were effectively imaged by suppressing only bone components and maintaining soft-tissue. These results suggest that the chances of missing abnormalities may be reduced by using the proposed method. The proposed method is useful for bone suppression in chest X-ray images.

Method for Catalytic Enantioselective Alkylation of Aldehydes Using Grignard Reagents as Alkyl Sources.

Alkyltitanium reagents, generated in situ from Grignard reagents and ClTi(O iPr)3, can be employed without further manipulation in the enantioselective alkylation of aldehyde by the catalysis of a chiral titanium complex derived from DTBP-H8-BINOL. The reaction is performed with good stoichiometry [1.5 equiv each of Grignard reagents and ClTi(O iPr)3] at a low catalyst loading (2 mol %), affording a variety of chiral secondary alcohols in high enantioselectivity and yields and, hence, realizing an asymmetric version of the Grignard reaction in an indirect manner.

Five-State Molecular Shuttling of a Pair of [2]Rotaxanes: Distinct Outputs in Response to Acid and Base Stimuli.

In this study we synthesized two acid-/base-controllable [2]rotaxanes featuring aminodiazobenzene and aminocoumarin units, respectively, as chromophores and dibenzo[24]crown-8 and dibenzo[25]crown-8 units, respectively, as their macrocyclic components. Each [2]rotaxane contained N-alkylarylamine (ammonium) and N,N-dialkylamine (ammonium) centers as binding sites for their crown ether components. The absorption patterns of the chromophores were dependent on the position of the encircling macrocyclic component and the degree of protonation, with three distinct states (under acidic, neutral, and basic conditions) evident for each [2]rotaxane. The mixed [2]rotaxane system displayed stepwise and independent molecular shuttling behavior based on the degree of protonation of the amino groups in response to both the amount and strength of added acids or bases; as such, the system provided five different absorption signals as outputs that could be read using UV/Vis spectroscopy.

The influence of outdoor thermal environment on young Japanese females.

The influence of short wave solar radiation appears to be strong outdoors in summer, and the influence of airflow appears to be strong outdoors in winter. The purpose of this paper was to clarify the influence of the outdoor environment on young Japanese females. This research shows the relationship between the physiological and psychological responses of humans and the enhanced conduction-corrected modified effective temperature (ETFe). Subjective experiments were conducted in an outdoor environment. Subjects were exposed to the thermal environment in a standing posture. Air temperature, humidity, air velocity, short wave solar radiation, long wave radiation, ground surface temperature, sky factor, and the green solid angle were measured. The temperatures of skin exposed to the atmosphere and in contact with the ground were measured. Thermal sensation and thermal comfort were measured by means of rating the whole-body thermal sensation (cold-hot) and the whole body thermal comfort (comfortable-uncomfortable) on a linear scale. Linear rating scales are given for the hot (100) and cold (0), and comfortable (100) and uncomfortable (0) directions only. Arbitrary values of 0 and 100 were assigned to each endpoint, the reported values read in, and the entire length converted into a numerical value with an arbitrary scale of 100 to give a linear rating scale. The ETFe considered to report a neither hot nor cold, thermally neutral sensation of 50 was 35.9 °C, with 32.3 °C and 42.9 °C, respectively, corresponding to the low and high temperature ends of the ETFe considered to report a neither comfortable nor uncomfortable comfort value of 50. The mean skin temperature considered to report a neither hot nor cold, thermally neutral sensation of 50 was 33.3 °C, with 31.0 °C and 34.3 °C, respectively, corresponding to the low and high temperature ends of the mean skin temperature considered to report a neither comfortable nor uncomfortable comfort value of 50. The acceptability raised the mean skin temperature even for thermal environment conditions in which ETFe was high.

Effect of the environmental stimuli upon the human body in winter outdoor thermal environment.

In order to manage the outdoor thermal environment with regard to human health and the environmental impact of waste heat, quantitative evaluations are indispensable. It is necessary to use a thermal environment evaluation index. The purpose of this paper is to clarify the relationship between the psychological thermal responses of the human body and winter outdoor thermal environment variables. Subjective experiments were conducted in the winter outdoor environment. Environmental factors and human psychological responses were measured. The relationship between the psychological thermal responses of the human body and the outdoor thermal environment index ETFe (enhanced conduction-corrected modified effective temperature) in winter was shown. The variables which influence the thermal sensation vote of the human body are air temperature, long-wave thermal radiation and short-wave solar radiation. The variables that influence the thermal comfort vote of the human body are air temperature, humidity, short-wave solar radiation, long-wave thermal radiation, and heat conduction. Short-wave solar radiation, and heat conduction are among the winter outdoor thermal environment variables that affect psychological responses to heat. The use of thermal environment evaluation indices that comprise short-wave solar radiation and heat conduction in winter outdoor spaces is a valid approach.

Three-state molecular shuttles operated using acid/base stimuli with distinct outputs.

This paper describes the acid/base-mediated three-state translational isomerization of two [2]rotaxanes, each containing N-alkylaniline and N,N-dialkylamine centers as binding sites for threaded dibenzo[24]crown-8 units. Under neutral conditions, the dialkylamine unit predominantly recognized the crown ether component through cooperative binding of a proton; when both amino units were protonated under acidic conditions, both translational isomers were generated; the addition of a strong base caused aniline-crown ether interactions to dominate. The three states of the [2]rotaxane featuring the 3,5-diphenylaniline terminus in its dumbbell-shaped component were accompanied by distinct absorptive outputs that were detectable using UV spectroscopy.

Identification of a novel NY-ESO-1 promiscuous helper epitope presented by multiple MHC class II molecules found frequently in the Japanese population.

NY-ESO-1 is a cancer-testis antigen that elicits strong cellular and humoral immune responses against NY-ESO-1-expressing tumors. Although CD4(+) T cells play a critical role in inducing antitumor immunity, little is known about MHC class II-restricted helper epitopes of the NY-ESO-1 antigen compared with MHC class I-restricted epitopes. Here, we searched for new NY-ESO-1 helper epitopes presented by MHC class II molecules, especially those found frequently in the Japanese population. We established five NY-ESO-1-specific helper T-cell lines from healthy Japanese donors using NY-ESO-1 recombinant protein and peptide. Using MHC class II-specific antibodies and a panel of Epstein-Barr virus-transformed B-cell lines, it was demonstrated that four out of the five T-cell lines recognized a region within NY-ESO-1(119-143) in the context of HLA-DRB1*0802, DRB1*0901, DRB1*1502 or DRB1*0405/*0410. In addition, using a set of overlapping 15-mer synthetic peptides, we found that NY-ESO-1(122-138) was a promiscuous region that bound to four distinct HLA-DR molecules found in the Japanese population. These findings expand the usefulness of NY-ESO-1 as a tool for tumor vaccine therapy in eliciting NY-ESO-1-specific helper T-cell responses, especially in Japanese cancer patients.

Natural killer T cells from interleukin-4-deficient mice are defective in early interferon-gamma production in response to alpha-galactosylceramide.

Discovery of the natural killer (NK) T cell-specific ligand, alpha-galactosylceramide (alpha-GalCer) has enabled us to investigate the functional regulation of NKT cells. However, the detailed mechanism of cytokine production by NKT cells remains to be elucidated. Here we evaluated the role of interleukin (IL)-4 in the production of interferon (IFN)-gamma from NKT cells using IL-4-deficient C57BL/6 mice (IL-4(-/-) mice). Administration of alpha-GalCer into wild-type C57BL/6 mice caused the production of both IFN-gamma and IL-4 in serum or cytoplasm within 4 h of the injection. Unexpectedly, however, IL-4(-/-) mice-derived NKT cells did not produce any IFN-gamma at early phase after primary stimulation with alpha-GalCer. Because NKT cells from IL-4(-/-) mice produced IFN-gamma when they were stimulated secondarily with alpha-GalCer in vitro for 72 h, NKT cells from IL-4(-/-) mice were not completely genetically deficient for IFN-gamma production. To elucidate which cells, NKT cells or dendritic cells (DC), were responsible for the deficiency in IFN-gamma production in IL-4(-/-) mice, we carried out an add-back experiment using purified NKT cells and DC, which were prepared from either wild-type mice or IL-4(-/-) mice. NKT cells from wild-type mice produced IFN-gamma when they were cocultured with DC prepared from either wild-type or IL-4(-/-) mice, whereas NKT cells from IL-4(-/-) mice did not produce IFN-gamma by coculturing with DC from either wild-type or IL-4(-/-) mice. These results indicate that NKT cells, not DC, were responsible for the deficiency in IFN-gamma production in IL-4(-/-) mice. Thus, IL-4 is required for the activation of NKT cells to produce IFN-gamma in response to alpha-GalCer.

AsialoGM1+CD8+ central memory-type T cells in unimmunized mice as novel immunomodulator of IFN-gamma-dependent type 1 immunity.

In unimmunized specific pathogen-free mice, there are unique memory-type CD8(+) T cell populations expressing asialoGM1 (ASGM1). These cells were classified into central memory-type T cells (T(CMT)) judging from their expression profile of CD44, IL-2Rbeta, CD62L and CCR7 cell-surface molecules. Among CD44(high)CD8(+) so-called memory CD8(+) T cell population, ASGM1(+)CD44(high)CD8(+) T(CMT), but not ASGM1(-)CD44(high)CD8(+) memory T cells, produced IFN-gamma by stimulation with anti-CD3 mAb. The physiological significance of ASGM1(+)CD8(+) T(CMT) as early source of IFN-gamma was also demonstrated in vivo. Namely, intravenous injection of anti-CD3 mAb (2 microg) resulted in early activation of IFN-gamma-producing ASGM1(+)CD8(+) T(CMT) cells as well as NKT and NK cells. Unexpectedly, however, few IFN-gamma-producing CD4(+) T cells were detected until 4 h after anti-CD3 mAb administration. Thus, ASGM1(+)CD8(+) T(CMT) were demonstrated to be early IFN-gamma producer, which may be crucial for T(h)1-dependent cellular immunity. Indeed, co-culture of naive CD4(+) T cells with ASGM1(+)CD8(+) T(CMT) but not ASGM1(-)CD8(+) T cells caused a great acceleration of IFN-gamma-producing T(h)1 cells in vitro. Finally, we found that T(h)1-prone C57BL/6 mice possessed higher percentage (10%) of ASGM1(+)CD8(+) T(CMT) in CD8(+) T cells compared with that (3%) of T(h)2-prone BALB/c mice. Moreover, ASGM1(+)CD8(+) T(CMT) derived from C57BL/6 mice produced higher levels of IFN-gamma compared with those from BALB/c mice. Thus, ASGM1(+)CD8(+) T(CMT), whose differentiation in vivo is genetically controlled, appear to play a critical role in the control of type 1 immunity, which is essential for therapy of tumors and infectious diseases.

Th1 cell adjuvant therapy combined with tumor vaccination: a novel strategy for promoting CTL responses while avoiding the accumulation of Tregs.

We have previously described a method for adoptive immunotherapy of cancer based on antigen-specific T(h)1 cells. However, efficient induction of anti-tumor responses using T(h)1 cells remains a formidable challenge, especially for MHC class II-negative tumors. In the present study, we sought to develop a novel strategy to eradicate established tumors of the MHC class II-negative, ovalbumin (OVA)-expressing EG-7 cells. Tumor-bearing mice were intradermally treated with OVA-specific T(h)1 cells, combined with the model tumor antigen (OVA), near the tumor-draining lymph node (DLN). We found that tumor growth was significantly inhibited by this strategy and approximately 50-60% of tumor-bearing mice were completely cured. Tumor eradication was crucially dependent on the generation of OVA/H-2K(b)-specific CTLs in the tumor DLNs and tumor site. The injected T(h)1 cells were mainly distributed in tumor DLNs, where they vigorously proliferated and enhanced the activation of dendritic cells. Strikingly, we also found that the accumulation of CD4(+)CD25(+) regulatory T cells (Tregs) was significantly inhibited in tumor DLNs by T(h)1 cell adjuvant therapy and this abrogation was associated with IFNgamma secreted by T(h)1 cells. These results identify T(h)1 cell adjuvant therapy combined with tumor vaccination as a novel approach to the treatment of human cancer.

Interleukin-12-responding asialoGM1+CD8+ central memory-type T cells as precursor cells for interferon-gamma-producing killer T cells.

While investigating CD8(+) memory T cells in unimmunized C57BL/6 mice, we found that there were unique memory-type CD8(+) T cells expressing asialoGM1 (ASGM1), CD62L and CCR7 cell surface molecules, which occupied approximately 10% of CD8(+) T cells and 35% of CD44(+) memory CD8(+) T cells. Culture of freshly isolated ASGM1(+)CD8(+) T cells with interleukin (IL)-12 plus IL-2 caused the proliferation and generation of killer T cells. Moreover, ASGM1(+)CD8(+) T cells, but not ASGM1(-)CD8(+) T cells, produced high levels of interferon (IFN)-gamma in response to IL-12 plus IL-2. Although ASGM1(+)CD8(+) T cells showed no significant responses to IL-12 alone or IL-2 alone, pulse incubation of ASGM1(+)CD8(+) T cells with IL-12 at an earlier time (0-12 h), and subsequently with IL-2 at a later time (12-24 h), caused the same levels of proliferation, killer cell generation and IFN-gamma production as when they were incubated simultaneously with IL-12 plus IL-2 for 24 h. Thus, ASGM1(+)CD8(+) T cells appeared to respond to IL-12 directly to acquire IL-2 responsiveness and differentiate into IFN-gamma-producing killer T cells. Indeed, freshly isolated ASGM1(+)CD8(+) T cells, but not ASGM1(-)CD8(+) T cells, expressed higher levels of IL-12R beta2 mRNA. The fact that IL-12 administration in vivo caused the generation of ASGM1(+)CD8(+) killer T cells in an IFN-gamma-dependent manner further indicated a physiological significance of ASGM1(+)CD8(+) central memory-type T cells in IL-12-induced immunoregulation for the therapy of tumors and infectious diseases.

Molecular design of glycoprotein mimetics: glycoblotting by engineered proteins with an oxylamino-functionalized amino acid residue.

The general and efficient method for the site-directed glycosylation of proteins is a key step in order to understand the biological importance of the carbohydrate chains of proteins and to control functional roles of the engineered glycoproteins in terms of the development of improved glycoprotein therapeutics. We have developed a novel method for site-directed glycosylation of proteins based on chemoselective blotting of common reducing sugars by genetically encoded proteins. The oxylamino-functionalized L-homoserine residues, 2-amino-4-O-(N-methylaminooxy) butanoic acid and 2-amino-4-aminooxy butanoic acid, were efficiently incorporated into proteins by using the four-base codon/anticodon pair strategy in Escherichia coli in vitro translation. Direct and chemoselective coupling between unmodified simple sugars and N-methylaminooxy group displayed on the engineered streptavidin allowed for the combinatorial synthesis of novel glycoprotein mimetics.

Convective heat transfer area of the human body.

In order to clarify the heat transfer area involved in convective heat exchange for the human body, the total body surface area of six healthy subjects was measured, and the non-convective heat transfer area and floor and chair contact areas for the following nine common body positions were measured: standing, sitting on a chair, sitting in the seiza position, sitting cross-legged, sitting sideways, sitting with both knees erect, sitting with a leg out, and the lateral and supine positions. The main non-convective heat transfer areas were: the armpits (contact between the upper arm and trunk regions), contact between the two legs, contacts between the fingers and toes, and contact between the hands and the body surface. Also, when sitting on the floor with some degree of leg contact (sitting in the seiza position, cross-legged, or sideways), there was a large non-convective heat transfer area on the thighs and legs. Even when standing or sitting in a chair, about 6-8% of the body surface did not transfer heat by convection. The results showed that the effective thermal convective area factor for the naked whole body in the standing position was 0.942. While sitting in a chair this factor was 0.860, while sitting in a chair but excluding the chair contact area it was 0.918, when sitting in the seiza position 0.818, when sitting cross-legged 0.843, in the sideways sitting position 0.855, when sitting with both knees erect 0.887, in the leg-out sitting position 0.906, while in the lateral position it was 0.877 and the supine position 0.844. For all body positions, the effective thermal convective area factor was greater than the effective thermal radiation area factor, but smaller than the total body surface area.