Glomerular Endothelial Cell Receptor ADGRF5 and the Integrity of the Glomerular Filtration Barrier.

BACKGROUND: Glomerular endothelial cells are recognized to be important for maintaining the glomerular filtration barrier. ADGRF5, an adhesion G protein-coupled receptor, has been suggested to be involved in endothelial cell function. However, the role of ADGRF5 in the glomerular filtration barrier integrity remains elusive. METHODS: Cellular expression of ADGRF5 in mouse glomerulus was determined by histological analyses. The impact of ADGRF5 deletion on the glomerular morphology, kidney function, and glomerular endothelial gene/protein expression was then analyzed using ADGRF5 knockout (Adgrf5-/-) mice and human primary glomerular endothelial cells. RESULTS: ADGRF5 was specifically expressed in the capillary endothelial cells within the glomerulus. Adgrf5-/- mice developed albuminuria and impaired kidney function with morphological defects in the glomeruli, namely glomerular hypertrophy, glomerular basement membrane splitting and thickening, diaphragmed fenestration and detachment of the glomerular endothelial cells, and mesangial interposition. These defects were accompanied by the altered expression of genes responsible for glomerular basement membrane organization (type IV collagens and laminins) and Krüppel-like factor 2 (Klf2) in glomerular endothelial cells. Moreover, ADGRF5 knockdown decreased COL4A3 and COL4A4 expression and increased KLF2 expression in human primary glomerular endothelial cells. CONCLUSIONS: The loss of ADGRF5 resulted in altered gene expression in glomerular endothelial cells, and perturbed the structure and permselectivity of the glomerular filtration barrier.

YIPF3 and YIPF4 regulate autophagic turnover of the Golgi apparatus.

The degradation of organelles by autophagy is essential for cellular homeostasis. The Golgi apparatus has recently been demonstrated to be degraded by autophagy, but little is known about how the Golgi is recognized by the forming autophagosome. Using quantitative proteomic analysis and two novel Golgiphagy reporter systems, we found that the five-pass transmembrane Golgi-resident proteins YIPF3 and YIPF4 constitute a Golgiphagy receptor. The interaction of this complex with LC3B, GABARAP, and GABARAPL1 is dependent on a LIR motif within YIPF3 and putative phosphorylation sites immediately upstream; the stability of the complex is governed by YIPF4. Expression of a YIPF3 protein containing a mutated LIR motif caused an elongated Golgi morphology, indicating the importance of Golgi turnover via selective autophagy. The reporter assays reported here may be readily adapted to different experimental contexts to help deepen our understanding of Golgiphagy.

Polygenic Risk Score, Cardiorespiratory Fitness, and Cardiometabolic Risk Factors: WASEDA'S Health Study.

PURPOSE: This study estimated an individual's genetic liability to cardiometabolic risk factors by polygenic risk score (PRS) construction and examined whether high cardiorespiratory fitness (CRF) modifies the association between PRS and cardiometabolic risk factors. METHODS: This cross-sectional study enrolled 1,296 Japanese adults aged ≥40 years. The PRS for each cardiometabolic trait (blood lipids, glucose, hypertension, and obesity) was calculated using the LDpred2 and clumping and thresholding methods. Participants were divided into low-, intermediate-, and high-PRS groups according to PRS tertiles for each trait. CRF was quantified as peak oxygen uptake (VO 2 peak) per kg body weight. Participants were divided into low-, intermediate-, and high-CRF groups according to the tertile VO 2 peak value. RESULTS: Linear regression analysis revealed a significant interaction between PRS for triglyceride (PRS TG ) and CRF groups on serum TG levels regardless of the PRS calculation method, and attenuated the association between PRS TG and TG levels in the high-CRF group. Logistic regression analysis revealed a significant sub-additive interaction between LDpred2 PRS TG and CRF on the prevalence of high TG, indicating that high CRF attenuated the genetic predisposition to high TG. Furthermore, a significant sub-additive interaction between PRS for body mass index and CRF on obesity was detected regardless of the PRS calculation method. These significant interaction effects on high TG and obesity were diminished in the sensitivity analysis using VO 2 peak per kg fat-free mass as the CRF index. Effects of PRSs for other cardiometabolic traits were not significantly attenuated in the high-CRF group regardless of PRS calculation methods. CONCLUSIONS: The findings of the present study suggest that individuals with high CRF overcome the genetic predisposition to high TG levels and obesity.

Associations between cardiorespiratory fitness and lifestyle-related factors with DNA methylation-based ageing clocks in older men: WASEDA'S Health Study.

DNA methylation-based age estimators (DNAm ageing clocks) are currently one of the most promising biomarkers for predicting biological age. However, the relationships between cardiorespiratory fitness (CRF), measured directly by expiratory gas analysis, and DNAm ageing clocks are largely unknown. We investigated the relationships between CRF and the age-adjusted value from the residuals of the regression of DNAm ageing clock to chronological age (DNAmAgeAcceleration: DNAmAgeAccel) and attempted to determine the relative contribution of CRF to DNAmAgeAccel in the presence of other lifestyle factors. DNA samples from 144 Japanese men aged 65-72 years were used to appraise first- (i.e., DNAmHorvath and DNAmHannum) and second- (i.e., DNAmPhenoAge, DNAmGrimAge, and DNAmFitAge) generation DNAm ageing clocks. Various surveys and measurements were conducted, including physical fitness, body composition, blood biochemical parameters, nutrient intake, smoking, alcohol consumption, disease status, sleep status, and chronotype. Both oxygen uptake at ventilatory threshold (VO2 /kg at VT) and peak oxygen uptake (VO2 /kg at Peak) showed a significant negative correlation with GrimAgeAccel, even after adjustments for chronological age and smoking and drinking status. Notably, VO2 /kg at VT and VO2 /kg at Peak above the reference value were also associated with delayed GrimAgeAccel. Multiple regression analysis showed that calf circumference, serum triglyceride, carbohydrate intake, and smoking status, rather than CRF, contributed more to GrimAgeAccel and FitAgeAccel. In conclusion, although the contribution of CRF to GrimAgeAccel and FitAgeAccel is relatively low compared to lifestyle-related factors such as smoking, the results suggest that the maintenance of CRF is associated with delayed biological ageing in older men.

Acute social jetlag augments morning blood pressure surge: a randomized crossover trial.

Although social jetlag (SJL) is generally considered a chronic condition, even acute SJL may have unfavorable effects on the cardiovascular system. We focused on the acute effects of SJL on morning blood pressure (BP) surge. This randomized crossover trial recruited 20 healthy men. In the SJL trial, participants delayed their bedtime by three hours on Friday and Saturday nights. Participants in the control (CON) trial implemented the same sleep-wake timing as on weekdays. Pre- and post-intervention measurements were performed to evaluate resting cardiovascular variables on Friday and Monday mornings, respectively. The ambulatory BP was automatically measured during the sleep and awake periods for 2 h after the participant woke up at night before pre- and post-intervention measurements. SJL (average mid-sleep time on weekends - average mid-sleep time on weekdays) occurred only in the SJL trial (SJL: 181 ± 24 min vs. CON: 8 ± 47 min). Carotid-femoral pulse wave velocity (cfPWV) and morning BP surge on Monday in the SJL trial were significantly higher than those on Friday in the SJL trial (cfPWV: P = 0.001, morning BP surge: P < 0.001), and those on Monday in the CON trial (cfPWV: P = 0.007; morning BP surge: P < 0.001). Furthermore, a significant positive correlation was found between ΔcfPWV and Δmorning BP surge (R = 0.587, P = 0.004). These results suggest that even acute SJL augments morning BP surge. This phenomenon may correspond to increased central arterial stiffness.State the details of Clinical Trials: Name: Effect of acute social jetlag on risk factors of lifestyle-related diseases. URL: https://center6.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000053204 . Unique identifier: UMIN000046639. Registration date: 17/01/2022.

Electronic Promotion of Methanol Synthesis over Cu-Loaded ZnO-Based Catalysts.

Methanol, a raw material for C1 chemistry, is industrially produced under harsh conditions using Cu/ZnO-based catalysts. The synthesis of methanol under mild conditions is a challenging subject using an improved catalyst. Here, Zn1-xSixO (ZSO) nanoparticles were synthesized by a thermal plasma method, and their work function and carrier concentration could be tuned by the Zn:Si ratio. The electrically conductive ZSO nanoparticles with a low work function enhanced the donation of electrons to loaded Cu and significantly promoted hydrogenation of CO to methanol, whereas insulating ZSO nanoparticles with a similar low work function did not. These results reveal that efficient electronic promotion by the transfer of electrons from a support to loaded Cu plays a key role in low-temperature methanol synthesis.

Changes in the skin characteristics associated with dehydration and rehydration.

The present study aimed to test the hypothesis that changes in the dermal tissue dielectric constant (TDC) and biomechanical properties of the skin would be correlated with the indicators related to dehydration. Ten healthy adult men were enrolled in three trials: no fluid intake (DEH), ad libitum fluid intake (AL-HYD), and programmed fluid intake (P-HYD) after exercise in a randomised crossover design. The participants performed a pedalling exercise at 60% heart rate reserve until 2% body mass loss. At 120 min after exercise, an incremental exercise test was carried out. Aerobic capacity, body composition, TDC, biomechanical properties of the skin (pliability, viscoelasticity, and total recovery), and indicators related to dehydration in the serum and urine were measured before and 120 min after exercise. Higher values of the pliability and viscoelasticity, and lower value of the total recovery on the hand were demonstrated in the P-HYD trial compared to the DEH trial (all P < 0.05). Changes in the TDC were significantly correlated with changes in body mass, total body water, serum osmolarity, and hematocrit (all P < 0.05). Changes in the biomechanical properties of the hand were significantly correlated with changes in body mass, hematocrit, and urine specific gravity (all P < 0.05). The present study showed that the changes in skin characteristics correlated with the body water and dehydration-associated indicators in the serum and urine, thus suggesting that skin characteristics may be useful in the assessment of dehydration.HighlightsThis study was the first to investigate the effect of dehydration and rehydration on the TDC and biomechanical properties of the skin upon instrumental measure, and not manual testing.This study confirmed the decline in aerobic capacity by dehydration and immediate recovery with sufficient rehydration.Changes in the TDC and the biomechanical properties of the skin correlated with the body water and dehydration-associated indicators in the serum and urine.Skin characteristics may be useful in the assessment of dehydration.

Muscle stretching induces the mechanoreflex response in human arterial blood pressure.

The muscle mechanoreflex has been considered to make a small contribution to the cardiovascular response to exercise in healthy humans because no pressor response has been observed during stimulation of mechanosensitive receptors, such as static passive stretching, during many human studies. There is room for rethinking this consideration since the pressor response to upper limb exercise is greater than that to lower limb exercise. We examined whether static passive stretching of the forearm muscles causes a muscle mechanoreflex-induced pressor response in humans. Eighteen healthy men were recruited for this study. After a 15-min rest period in the supine position with a neutral (0°) wrist joint angle, all participants completed static passive stretching of the forearm for 60 s at four different intensities: minimal painful passive stretching (PPS), moderate-intensity passive stretching (MPS), low-intensity passive stretching (LPS), and no load (NL). During the procedure, beat-to-beat arterial blood pressure was measured using finger photoplethysmography. The force generated between the passively stretched hand and the experimenter's hands was recorded using a force transducer. Mean arterial pressure (MAP) during PPS and MPS significantly increased from baseline during the last 40 s (P < 0.05). MAP was significantly greater at 50 s and 60 s, depending on the intensity. MPS induced a greater peak response in MAP than lower intensities (P < 0.05). None of the subjects reported pain during the MPS and LPS trials. Static passive stimulation of the forearm is an effective method of isolating the muscle mechanoareflex-induced pressor response in humans.NEW & NOTEWORTHY The muscle mechanoreflex was considered to have a small contribution to cardiovascular regulation during exercise in healthy humans. In contrast, the results of this study indicate that static stretching of the forearm induces a pressor response in healthy humans and suggest that the mechanoreflex explicitly induces the pressor response during exercise in humans. The methods applied are useful for evaluating the pressor response to the mechanoreflex regardless of health, aging, and disease.

Effects of Intestinal Bacterial Hydrogen Gas Production on Muscle Recovery following Intense Exercise in Adult Men: A Pilot Study.

This study aimed to examine the effects of hydrogen gas (H2) produced by intestinal microbiota on participant conditioning to prevent intense exercise-induced damage. In this double-blind, randomized, crossover study, participants ingested H2-producing milk that induced intestinal bacterial H2 production or a placebo on the trial day, 4 h before performing an intense exercise at 75% maximal oxygen uptake for 60 min. Blood marker levels and respiratory variables were measured before, during, and after exercise. Visual analog scale scores of general and lower limb muscle soreness evaluated were 3.8- and 2.3-fold higher, respectively, on the morning after treatment than that before treatment during the placebo trial, but not during the test beverage consumption. Urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) concentrations and production rates significantly increased with placebo consumption; no changes were observed with test beverage consumption. After exercise, relative blood lactate levels with H2-producing milk consumption were lower than those with placebo consumption. A negative correlation was observed between the variation of 8-OHdG and the area under the curve (AUC) of breath H2 concentrations. Lipid oxidation AUC was 1.3-fold higher significantly with H2-producing milk than with placebo consumption. Conclusively, activating intestinal bacterial H2 production by consuming a specific beverage may be a new strategy for promoting recovery and conditioning in athletes frequently performing intense exercises.

Muscle stiffening is associated with muscle mechanoreflex-mediated cardioacceleration.

PURPOSE: Although the muscle mechanoreflex is an important mediator to cardiovascular regulation during exercise, its modulation factors remain relatively unknown. Therefore, the purpose of this study was to investigate the effect of muscle stiffness on the muscle mechanoreflex. METHODS: Participants were divided based on their median muscle stiffness (2.00 Nm/mm) into a low group (n = 15) and a high group (n = 15), and the muscle mechanoreflex was compared between the groups. After a 15-min rest in the supine position, heart rate (HR), blood pressure (BP), stroke volume (SV), and cardiac output (CO) were measured at rest for 3 min and during static passive dorsiflexion (SPD) at 20° for 1 min. Following a 15-min re-rest, muscle stiffness and passive resistive torque were evaluated in the distal end of the muscle belly of the medial gastrocnemius. RESULTS: Peak relative changes in HR (low group: 6 ± 4% and high group: 12 ± 4%) and CO (low group: 8 ± 10% and high group: 13 ± 9%) were greater in the high group than in the low group (both, P < 0.05). A significant positive correlation was found between resistive torque during SPD and muscle stiffness and peak relative changes in HR (r = 0.51 and 0.61, both P < 0.05). However, there was no correlation between muscle elongation during SPD and peak relative changes in HR (r = - 0.23, P = 0.20). CONCLUSION: These findings suggest that muscle stiffness may be modulatory factor of muscle mechanoreflex.

Impact of acute dynamic exercise on vascular stiffness in the retinal arteriole in healthy subjects.

Acute exercise can improve vascular stiffness in the conduit artery, but its effect on the retinal arterioles is unknown. The present study investigated the effects of acute dynamic exercise on retinal vascular stiffness. In experiment 1, we measured the cardio-ankle vascular index (CAVI), carotid artery intima-media thickness (carotid IMT), and retinal blood velocity by laser speckle flowgraphy in 28 healthy old and 28 young men (69 ± 3 and 23 ± 3 yr, respectively). Pulse waveform variables, which were used as an index of retinal vascular stiffness, were assessed by retinal blood flow velocity profile analysis. In experiment 2, 18 healthy old and 18 young men (69 ± 3 and 23 ± 3 yr, respectively) underwent assessment of pulse waveform variables after a 30-min bout of moderate cycling exercise at an intensity of 60% heart rate reserve. There was a significant difference in the baseline pulse waveform variables between the old and young groups. Pulse waveform variables in the retinal arteriole did not significantly change after acute dynamic exercise, whereas CAVI significantly decreased. These findings suggest that retinal vascular stiffness does not change by acute exercise. The effect of exercise on vascular stiffness in the retinal arterioles might be different from that in the conduit artery.NEW & NOTEWORTHY Acute dynamic exercise is well known to improve vascular stiffness in the conduit artery while its effect on the retinal arterioles has been unknown. This study showed that an acute dynamic exercise does not change vascular stiffness in the retinal arteriole in healthy humans. Different responses to acute dynamic exercise in vascular stiffness in retinal arterioles and conduit arteries are suggested.

Carbonic anhydrase inhibitor induces otic hair cell apoptosis via an intrinsic pathway and ER stress in zebrafish larvae.

Carbonic anhydrase (CA) catalyzes reversible hydration of CO2 to HCO3 - to mediate pH and ion homeostasis. Some chemical pollutants have been reported to have inhibitory effects on fish CA. In this study, we investigated effects of a CA inhibitor ethoxyzolamide (EZA) on neuromasts development during zebrafish embryogenesis, since embryogenesis in aquatic organisms can be particularly sensitive to water pollution. EZA caused alteration of pH and calcium concentration and production of reactive oxygen species (ROS) in larvae, and induced apoptosis in hair cells especially in the otic neuromast, in which CA2 was distributed on the body surface. mRNA levels of apoptotic genes and caspase activities were increased by EZA, whereas anti-oxidants and apoptotic inhibitors, Bax, NF-κB, and p53 inhibitors significantly relieved the induction of hair cell death. Also, mRNA levels of Bip and CHOP, which are induced in response to ER stress, were upregulated by EZA, suggesting that EZA induces otic hair cell apoptosis via the intrinsic mitochondrial pathway and ER stress. Our results demonstrated an essential role of CA in neuromast development via maintenance of ion transport and pH, and that the CA, which is directly exposed to the ambient water, shows marked sensitivity to EZA.

Effects of Greater Central Arterial Stiffness on Cardiovagal Baroreflex Sensitivity in Resistance-Trained Men.

BACKGROUND: Compared with age-matched untrained men, resistance-trained men who have undergone long duration training (> 2 years) at a high frequency (> 5 days/week) may be lower cardiovagal baroreflex sensitivity (BRS) because of central arterial stiffening. Therefore, the purpose of this study was to examine the effect of greater central arterial stiffness in resistance-trained men on cardiovagal BRS in a cross-sectional study to compare resistance-trained men with age-matched untrained men. METHODS: This cross-sectional study included resistance-trained men (n = 20; age: 22 ± 3; body mass index: 26.7 ± 2.2) and age-matched untrained men (control group: n = 20; age: 25 ± 2; body mass index: 23.7 ± 2.4). The ß-stiffness index and arterial compliance were assessed at the right carotid artery using a combination of a brightness mode ultrasonography system for the carotid artery diameter and applanation tonometry for the carotid blood pressure. And, the cardiovagal BRS was estimated by the slope of the R-R interval and systolic blood pressure during Phase II and IV of Valsalva maneuver (VM). The participants maintained an expiratory mouth pressure of 40 mmHg for 15 s in the supine position. RESULTS: The ß-Stiffness index was significantly higher in the resistance-trained group than in the control group (5.9 ± 1.4 vs. 4.4 ± 1.0 a.u., P < 0.01). In contrast, the resistance-trained group had significantly lower arterial compliance (0.15 ± 0.05 vs. 0.20 ± 0.04 mm2/mmHg, P < 0.01) and cardiovagal BRS during Phase IV of VM (9.0 ± 2.5 vs. 12.9 ± 5.4 ms/mmHg, P < 0.01) than the control group and. Moreover, cardiovagal BRS during Phase IV of VM was inversely and positively correlated with the ß-stiffness index (r = - 0.59, P < 0.01) and arterial compliance (r = 0.64, P < 0.01), respectively. CONCLUSION: Resistance-trained group had greater central arterial stiffness and lower cardiovagal BRS Phase IV compared with control group. Moreover, the central arterial stiffening was related to cardiovagal BRS Phase IV. These results suggest that greater central arterial stiffness in resistance-trained men may be associated with lower cardiovagal BRS. Trial Registration University hospital Medical Information Network (UMIN) in Japan, UMIN000038116. Registered on September 27, 2019.

Determinants of Resting Oxidative Stress in Middle-Aged and Elderly Men and Women: WASEDA'S Health Study.

Previous studies have not investigated the determinants of resting oxidative stress, including physical fitness, as it relates to redox regulation. The present study therefore was aimed at identifying lifestyle and biological factors that determine resting oxidative stress, including objectively measured physical fitness. In 873 middle-aged and elderly men and women, age and anthropometric parameters, lifestyle-related parameters, medication and supplementation status, physical fitness, biochemical parameters, and nutritional intake status, as well as three plasma oxidative stress markers: protein carbonyl (PC), F2-isoprostane (F2-IsoP), and thiobarbituric acid reactive substances (TBARS), were surveyed and measured. The determinants of PC, F2-IsoP, and TBARS in all participants were investigated using stepwise multiple regression analysis. In PC, age (ß = -0.11, P = 0.002), leg extension power (ß = -0.12, P = 0.008), BMI (ß = 0.12, P = 0.004), and HDL-C (ß = 0.08, P = 0.040) were included in the regression model (adjusted R 2 = 0.018). In the F2-IsoP, smoking status (ß = 0.07, P = 0.060), BMI (ß = 0.07, P = 0.054), and HbA1c (ß = -0.06, P = 0.089) were included in the regression model (adjusted R 2 = 0.006). In TBARS, glucose (ß = 0.18, P < 0.001), CRF (ß = 0.16, P < 0.001), age (ß = 0.15, P < 0.001), TG (ß = 0.11, P = 0.001), antioxidant supplementation (ß = 0.10, P = 0.002), and HbA1c (ß = -0.13, P = 0.004) were included in the regression model (adjusted R 2 = 0.071). In conclusion, the present study showed that age, anthropometric index, lifestyle-related parameters, medication and supplementation status, objectively measured physical fitness, biochemical parameters, and nutritional intake status explain less than 10% of oxidative stress at rest.

The Effects of Beverage Intake after Exhaustive Exercise on Organ Damage, Inflammation and Oxidative Stress in Healthy Males.

Strenuous exercise induces organ damage, inflammation and oxidative stress. To prevent exercise-induced organ damage, inflammation and oxidative stress, rehydrating may be an effective strategy. In the present study, we aimed to examine whether beverage intake after exhaustive exercise to recover from dehydration prevents such disorders. Thirteen male volunteers performed incremental cycling exercise until exhaustion. Immediately after exercise, the subjects drank an electrolyte containing water (rehydrate trial: REH) or did not drink any beverage (control trial: CON). Blood samples were collected before (Pre), immediately (Post), 1 h and 2 h after exercise. Urine samples were also collected before (Pre) and 2 h after exercise. We measured biomarkers of organ damage, inflammation and oxidative stress in blood and urine. Biomarkers of muscle, renal and intestinal damage and inflammation increased in the blood and urine after exercise. However, changes in biomarkers of organ damage and inflammation did not differ between trials (p > 0.05). The biomarker of oxidative stress, thiobarbituric acid reactive substances (TBARS), in plasma, showed different changes between trials (p = 0.027). One hour after exercise, plasma TBARS concentration in REH had a higher trend than that in CON (p = 0.052), but there were no significant differences between Pre and the other time points in each trial. These results suggest that beverage intake after exercise does not attenuate exercise-induced organ damage, inflammation or oxidative stress in healthy males. However, rehydration restores exercise-induced oxidative stress more quickly.

Orphan GPR116 mediates the insulin sensitizing effects of the hepatokine FNDC4 in adipose tissue.

The proper functional interaction between different tissues represents a key component in systemic metabolic control. Indeed, disruption of endocrine inter-tissue communication is a hallmark of severe metabolic dysfunction in obesity and diabetes. Here, we show that the FNDC4-GPR116, liver-white adipose tissue endocrine axis controls glucose homeostasis. We found that the liver primarily controlled the circulating levels of soluble FNDC4 (sFNDC4) and lowering of the hepatokine FNDC4 led to prediabetes in mice. Further, we identified the orphan adhesion GPCR GPR116 as a receptor of sFNDC4 in the white adipose tissue. Upon direct and high affinity binding of sFNDC4 to GPR116, sFNDC4 promoted insulin signaling and insulin-mediated glucose uptake in white adipocytes. Indeed, supplementation with FcsFNDC4 in prediabetic mice improved glucose tolerance and inflammatory markers in a white-adipocyte selective and GPR116-dependent manner. Of note, the sFNDC4-GPR116, liver-adipose tissue axis was dampened in (pre) diabetic human patients. Thus our findings will now allow for harnessing this endocrine circuit for alternative therapeutic strategies in obesity-related pre-diabetes.

ppGpp functions as an alarmone in metazoa.

Guanosine 3',5'-bis(pyrophosphate) (ppGpp) functions as a second messenger in bacteria to adjust their physiology in response to environmental changes. In recent years, the ppGpp-specific hydrolase, metazoan SpoT homolog-1 (Mesh1), was shown to have important roles for growth under nutrient deficiency in Drosophila melanogaster. Curiously, however, ppGpp has never been detected in animal cells, and therefore the physiological relevance of this molecule, if any, in metazoans has not been established. Here, we report the detection of ppGpp in Drosophila and human cells and demonstrate that ppGpp accumulation induces metabolic changes, cell death, and eventually lethality in Drosophila. Our results provide the evidence of the existence and function of the ppGpp-dependent stringent response in animals.

Tape stripping method is useful for the quantification of antimicrobial peptides on the human skin surface including the stratum corneum.

Antimicrobial peptides (AMPs) play an important role in innate immunity in human skin. It is known that AMPs mainly function in the stratum corneum. Therefore, AMP concentrations in the stratum corneum need to be precisely measured to clarify functional and physiological importance of AMPs in cutaneous defence. Tape stripping (TS) is a well-established method by which components in the stratum corneum can be collected. However, the usefulness of the TS method for measuring AMP concentration in human skin remains unclear. Therefore, we compared it with another popular method, skin rinsing, which had been established as a method for measuring AMP concentration in human skin. When investigated on healthy medial forearm using RNase 7, which is one of the typical AMPs, as an index, there was a significant positive correlation between RNase 7 concentrations measured by the TS method at adjacent forearm sites, demonstrating the reproducibility of the TS method. Next, a significant positive correlation was detected in RNase 7 concentrations measured using the TS and the skin rinsing method, indicating that the TS method is comparable to the skin rinsing method. Thus, we speculate that the TS method is useful for measuring AMP concentration in human skin.

Characteristics and Functions of the Yip1 Domain Family (YIPF), Multi-Span Transmembrane Proteins Mainly Localized to the Golgi Apparatus.

Yip1 domain family (YIPF) proteins are multi-span, transmembrane proteins mainly localized in the Golgi apparatus. YIPF proteins have been found in virtually all eukaryotes, suggesting that they have essential function(s). Saccharomyces cerevisiae contains four YIPFs: Yip1p, Yif1p, Yip4p, and Yip5p. Early analyses in S. cerevisiae indicated that Yip1p and Yif1p bind to each other and play a role in budding of transport vesicles and/or fusion of vesicles to target membranes. However, the molecular basis of their functions remains unclear. Analysis of YIPF proteins in mammalian cells has yielded significant clues about the function of these proteins. Human cells have nine family members that appear to have overlapping functions. These YIPF proteins are divided into two sub-families: YIPFα/Yip1p and YIPFß/Yif1p. A YIPFα molecule forms a complex with a specific partner YIPFß molecule. In the most broadly hypothesized scenario, a basic tetramer complex is formed from two molecules of each partner YIPF protein, and this tetramer forms a higher order oligomer. Three distinct YIPF protein complexes are formed from pairs of YIPFα and YIPFß proteins. These are differently localized in either the early, middle, or late compartments of the Golgi apparatus and are recycled between adjacent compartments. Because a YIPF protein is predicted to have five transmembrane segments, a YIPF tetramer complex is predicted to have 20 transmembrane segments. This high number of transmembrane segments suggests that YIPF complexes function as channels, transporters, or transmembrane receptors. Here, the evidence from functional studies of YIPF proteins obtained during the last two decades is summarized and discussed.