Outdoor Scene Classrooms to Arrest Myopia: Design and Baseline Characteristics.

PURPOSE: This study aimed to investigate the impact on childhood myopia of classrooms with spatial properties of classrooms resembling those of outdoor environments. This article describes the design, baseline characteristics, and the acceptability of this strategy. METHODS: Classrooms had custom-made wallpaper installed with forest and sky scenes that had spatial frequency spectra comparable with outdoor environments (i.e., outdoor scene classrooms). Acceptability of this strategy was evaluated by questionnaires. Outcomes to access the efficacy include cumulative proportion of myopia, change of cycloplegic spherical equivalent refractive error, and axial length. RESULTS: Ten classes, comprising 520 students, were randomly assigned into outdoor scene or tradition classrooms. There was no difference in refractive status between two groups (myopia/emmetropia/hyperopia, 16.3% vs. 49.4% vs. 34.2% in outdoor scene classrooms, 18.3% vs. 49.0% vs. 32.7% in traditional classrooms; P = .83). Compared with the traditional classrooms, 88.9% of teachers and 87.5% of students felt the outdoor scene classrooms enjoyable, 22.2% of teachers and 75.3% of students reported higher concentration, and 77.8% of teachers and 15.2% of students reported no change. In addition, 44.4% of teachers and 76.0% of students reported higher learning efficiency in the outdoor scene classrooms, and 55.6% of teachers and 18.3% of students reported no change. CONCLUSIONS: Outdoor scene classrooms are appealing to teachers and students. Outcomes of the study will inform the efficacy of this strategy in Chinese children.

Prediction model of gonadotropin starting dose and its clinical application in controlled ovarian stimulation.

BACKGROUND: Selecting an appropriate and personalized Gn starting dose (GSD) is an essential procedure for determining the quality and quantity of oocytes in the controlled ovarian stimulation (COS) process of the in-vitro fertilization (IVF) treatment cycle. The current approach for determining the GSD is mainly based on the experience of a clinician, lacking unified and scientific standards. This study aims to establish a prediction model of GSD, based on which good COS outcomes can be achieved with the influencing factors comprehensively evaluated quantitatively. MATERIAL AND METHODS: We collected a total of 1555 patients undergoing the first oocytes retrieving cycle and conducted correlation analysis to find the significant factors related to the GSD. Two GSD models are built based on two popular machine learning approaches, and the one with better model performance is selected as the final model. Finally, clinical application and validation were conducted to verify the effectiveness of the proposed model. RESULTS: (1) Age, duration of infertility, type of infertility, body mass index (BMI), antral follicle count (AFC), basal follicle stimulating hormone (bFSH), estradiol (E2), luteinizing hormone (LH), anti-Müllerian hormone (AMH) and COS treatment regimen were closely related to the GSD (P < 0.05). (2) The selected model has good modeling performance in terms of both root mean square error (RMSE) (29.87 ~ 34.21) and regression coefficient R (0.947 ~ 0.953). (3) A comprehensive evaluation of influencing factors for GSD is conducted and shows that the top four most significant factors are age, AMH, AFC, and BMI. (4) The proposed GSD can approximate the actual value well in the clinical application, with the mean absolute error of only 11.26 units, and the recommended results can prompt the number of oocytes retrieved (NOR) close to the optimal number. CONCLUSION: Modeling the GSD value with machine learning approaches is feasible and effective, and the proposed model has good clinical application for determining the GSD in the IVF treatment cycle.

A dynamic and multilocus metabolic regulation strategy using quorum-sensing-controlled bacterial small RNA.

Metabolic regulation strategies have been developed to redirect metabolic fluxes to production pathways. However, it is difficult to screen out target genes that, when repressed, improve yield without affecting cell growth. Here, we report a strategy using a quorum-sensing system to control small RNA transcription, allowing cell-density-dependent repression of target genes. This strategy is shown with convenient operation, dynamic repression, and availability for simultaneous regulation of multiple genes. The parameters Ai, Am, and RA (3-oxohexanoyl-homoserine lactone [AHL] concentrations at which half of the maximum repression and the maximum repression were reached and value of the maximum repression when AHL was added manually, respectively) are defined and introduced to characterize repression curves, and the variant LuxRI58N is identified as the most suitable tuning factor for shake flask culture. Moreover, it is shown that dynamic overexpression of the Hfq chaperone is the key to combinatorial repression without disruptions on cell growth. To show a broad applicability, the production titers of pinene, pentalenene, and psilocybin are improved by 365.3%, 79.5%, and 302.9%, respectively, by applying combinatorial dynamic repression.

Comparative Morphology of the Lungs and Skin of two Anura, Pelophylax nigromaculatus and Bufo gargarizans.

The lungs and skin are important respiratory organs in Anura, but the pulmonary structure of amphibians remains unclear due to the lack of a suitable procedure. This study improved the procedure used for fixing lungs tissues and used light microscopy, transmission electron microscopy and scanning electron microscopy to reveal the differences in the lung and skin morphologies between Pelophylax nigromaculatus (P. nigromaculatus) and Bufo gargarizans (B. gargarizans). In P. nigromaculatus and B. gargarizans, the cystic lungs comprise a continuous outer pulmonary wall on which primary, secondary, and tertiary septa attach, and a number of regular lattices form from raised capillaries and the pulmonary epithelium on the surfaces of the pulmonary wall and septa. Each lattice in P. nigromaculatus consists of several elliptical sheets and flat bottom, and the septa are distributed with denser sheets and have a larger stretching range than the pulmonary wall. The lattice in B. gargarizans consists of thick folds and an uneven bottom with several thin folds, and the septa have more developed thick and thin folds than the pulmonary wall. However, the density of the pulmonary microvilli, the area of a single capillary, the thicknesses of the blood-air barrier, pulmonary wall and septum, and the lung/body weight percentage obtained for B. gargarizans were higher than those found for P. nigromaculatus. In P. nigromaculatus, the dorsal skin has dense capillaries and a ring surface structure with mucus layer on the stratum corneum, and the ventral skin is slightly keratinized. In B. gargarizans, the stratum corneum in both the dorsal and ventral skins is completely keratinized. A fine ultrastructure analysis of P. nigromaculatus and B. gargarizans revealed that the pulmonary septa are more developed than the pulmonary walls, which means that the septa have a stronger respiratory function. The more developed lungs are helpful for the adaptation of B. gargarizans to drought environments, whereas P. nigromaculatus has to rely on more vigorous skin respiration to adapt to a humid environment.

Identification of an Upper Limit of Tumor Burden for Downstaging in Candidates with Hepatocellular Cancer Waiting for Liver Transplantation: A West-East Collaborative Effort.

Abstract: Since the introduction of Milan Criteria, all scoring models describing the prognosis of hepatocellular cancer (HCC) after liver transplantation (LT) have been exclusively based on characteristics available at surgery, therefore neglecting the intention-to-treat principles. This study aimed at developing an intention-to-treat model through a competing-risk analysis. Using data available at first referral, an upper limit of tumor burden for downstaging was identified beyond which successful LT becomes an unrealistic goal. Twelve centers in Europe, United States, and Asia (Brussels, Sapienza Rome, Padua, Columbia University New York, Innsbruck, Medanta-The Medicity Dehli, Hong Kong, Kyoto, Kaohsiung Taiwan, Mainz, Fukuoka, Shulan Hospital Hangzhou) created a Derivation (n = 2318) and a Validation Set (n = 773) of HCC patients listed for LT between January2000-March 2017. In the Derivation Set, the competing-risk analysis identified two independent covariables predicting post-transplant HCC-related death: combined HCC number and diameter (SHR = 1.15; p < 0.001) and alpha-fetoprotein (AFP) (SHR = 1.80; p < 0.001). WE-DS Model showed good diagnostic performances at internal and external validation. The identified upper limit of tumor burden for downstaging was AFP ≤ 20 ng/mL and up-to-twelve as sum of HCC number and diameter; AFP = 21-200 and up-to-ten; AFP = 201-500 and up-to-seven; AFP = 501-1000 and up-to-five. The WE-DS Model proposed here, based on morphologic and biologic data obtained at first referral in a large international cohort of HCC patients listed for LT, allowed identifying an upper limit of tumor burden for downstaging beyond which successful LT, following downstaging, results in a futile transplantation.

[Prospective cases-control study on arthroscopic for the treatment of ischial tuberosity cyst].

OBJECTIVE: To compare technique and clinical effect of arthroscopic and conventional sectional therapy for the treatment of ischial tuberosity cyst. METHODS: From May 2014 and September 2016, 49 patients with symptomatic ischial tuberosity cyst were randomized divided into arthroscopic group and conventional section group by envelope method. There were 24 patients in arthroscopic group, including 16 males and 8 females aged from 42 to 81years old with an average age of (64.1±9.3) years old;the courses of disease ranged from 2 to 36 months with an average of (17.0±9.1) months;treated with removing cyst wall under arthroscopic and build artificial lacuna around cyst. There were 25 patients in conventional section group, including 11 males and 14 females aged from 47 to 79 years old with an average of (61.2±10.6) years old; the courses of disease ranged from 4 to 36 months with an average of (17.5±8.5) months;treated with cutting off lump with transverse incision. Operative time, blood loss, fluid volume, hospital stays and postoperative complication were observed and compared, VAS score were applied to evaluate pain degree. RESULTS: Forty-nine patients were followed up from 6 to 18 months with an average of (11.3±3.3) months. In conventional group, 2 patients occurred incision infection, 1 case reoccurred. All patients in arthroscopic group were healed at stage I. Operative time, blood loss, fluid volume, and hospital stays in arthroscopic group were (54.7±7.7) min, (20.8±3.5) ml, (20.3±5.6) ml, (2.8±0.6) d respectively and better than that of conventional group(71.8±8.8) min, (67.3±12.0) ml, (103.6±20.3) ml, (7.8±2.9) d. Postoperative VAS score in arthroscopic group on the first day, seventh days and first month were 2.6±0.7, 0.5±0.6, 0.3±0.5, and significantly lower than that of conventional group 6.0±0.7, 3.0±1.0, 1.1±1.0 respectively(P<0.05). There were no statistical difference in postoperative complications between two groups(P>0.05). CONCLUSIONS: Compared with conventional group, arthroscopic for ischial tuberosity cyst has advantages of minimal invasive, less blood loss during perioperative period, less pain degree after surgery, safety and rapid recovery. But arthroscopic skills are needed for surgeons. It should be recommended in clinical practice.

SopB-Mediated Recruitment of SNX18 Facilitates Salmonella Typhimurium Internalization by the Host Cell.

To invade epithelial cells, Salmonella enterica serovar Typhimurium (S. Typhimurium) induces macropinocytosis through the action of virulence proteins delivered across the host cell membrane via a type III secretion system. We show that after docking at the plasma membrane S. Typhimurium triggers rapid recruitment of cytosolic SNX18, a SH3-PX-BAR domain sorting nexin protein, to the bacteria-induced membrane ruffles and to the nascent Salmonella-containing vacuole. SNX18 recruitment required the inositol-phosphatase activity of the Salmonella effector SopB and an intact phosphoinositide-binding site within the PX domain of SNX18, but occurred independently of Rho-GTPases Rac1 and Cdc42 activation. SNX18 promotes formation of the SCV from the plasma membrane by acting as a scaffold to recruit Dynamin-2 and N-WASP in a process dependent on the SH3 domain of SNX18. Quantification of bacteria uptake revealed that overexpression of SNX18 increased bacteria internalization, whereas a decrease was detected in cells overexpressing the phosphoinositide-binding mutant R303Q, the ΔSH3 mutant, and in cells where endogenous levels of SNX18 were knocked-down. This study identifies SNX18 as a novel target of SopB and suggests a mechanism where S. Typhimurium engages host factors via local manipulation of phosphoinositide composition at the site of invasion to orchestrate the internalization process.

[The pedicled anterolateral thigh flap for penile reconstruction].

OBJECTIVE: To introduce the application of pedicled anterolateral thigh (ALT) flap for total penile reconstruction and to investigate its feasibility and effect. METHODS: From May 2011 to May 2015, 12 male patients presented with absence of the penis or congenital malformation received phalloplasty with the pedicled ALT flap. Of them, the median age was 35 years old (range, 20-57 years). The size of the flaps ranged from 11 cm x 11 cm to 12 cm x 15 cm. 8 patients underwent urethra reconstruction with tube-in-tube flaps and other 4 patients with scrotal septal flaps. In this series, we performed one-stage urethral anastomosis in 4 cases and second-stage urethral anastomosis 6 months after the phalloplasty in 8 cases. RESULTS: An acceptable reconstructed phallus was achieved in 10 patients. These flaps were primarily healed with satisfactory functional and cosmetic results. The reconstructed penis was completely necrosis because of the flap failure in one case. The distal half of the phallus was lost due to infection in one case. Both of them were treated with pedicled ALT flap from the other side. Reliable results were achieved. At a median follow-up of 1.5 year (range 1 to 4 years), 9 patients (90%) were fully satisfied with phallic cosmesis and size, and 4 patients who was married had successful sexual intercourse. 8 patients had normal urinate function. Fistula was developed in 2 patients (20%) which was treated with delayed repair at 6-12 months with local scrotal flaps. CONCLUSIONS: The pedicled ALT flap can be simply used to reconstruct an entire penis as well as a urethra. It has several advantages including a less conspicuous donor site, greater bulk, better color match and no necessary for microsurgery.

[Reconstruction of facial soft tissue defects with pedicled expanded flaps].

OBJECTIVE: To investigate the application of pedicled expanded flaps for the reconstruction of facial soft tissue defects. METHODS: The expanded skin flaps, pedicled with orbicularis oculi muscle, submental artery, the branch of facial artery, superficial temporal artery, interior upper arm artery, had similar texture and color as facial soft tissue. The pedicled expanded flaps have repaired the facial soft tissue defects. RESULTS: Between Jan. 2003 to Dec. 2013, 157 cases with facial soft tissue defects were reconstructed by pedicled expanded flaps. Epidermal necrosis happened at the distal end of 8 expanded flaps, pedicled with interior upper arm artery(4 cases), orbicularis oculi muscle(3 cases) and submental artery(1 case), which healed spontaneously after dressing. All the other flaps survived completely with similar color and inconspicuous scar. 112 cases were followed up for 8 months to 8 years. Satisfactory results were achieved in 75 cases. 37 cases with hypertrophic scar at incisions need secondary operation. CONCLUSIONS: Island pedicled expanded flap with similar texture and color as facial soft tissue is suitable for facial soft tissue defects. The facial extra-incision and large dog-ear deformity could be avoided.

The Vps35 D620N mutation linked to Parkinson's disease disrupts the cargo sorting function of retromer.

The retromer is a trimeric cargo-recognition protein complex composed of Vps26, Vps29 and Vps35 associated with protein trafficking within endosomes. Recently, a pathogenic point mutation within the Vps35 subunit (D620N) was linked to the manifestation of Parkinson's disease (PD). Here, we investigated details underlying the molecular mechanism by which the D620N mutation in Vps35 modulates retromer function, including examination of retromer's subcellular localization and its capacity to sort cargo. We show that expression of the PD-linked Vps35 D620N mutant redistributes retromer-positive endosomes to a perinuclear subcellular localization and that these endosomes are enlarged in both model cell lines and fibroblasts isolated from a PD patient. Vps35 D620N is correctly folded and binds Vps29 and Vps26A with the same affinity as wild-type Vps35. While PD-linked point mutant Vps35 D620N interacts with the cation-independent mannose-6-phosphate receptor (CI-M6PR), a known retromer cargo, we find that its expression disrupts the trafficking of cathepsin D, a CI-M6PR ligand and protease responsible for degradation of α-synuclein, a causative agent of PD. In summary, we find that the expression of Vps35 D620N leads to endosomal alterations and trafficking defects that may partly explain its action in PD.

Decatantalate--the last member of the group 5 decametalate family.

A tetra-n-butylammonium (TBA) salt of [Ta10O28](6-) was synthesized by heating TBA6[H2Ta6O19] in toluene for a prolonged period. X-ray structural analysis of TBA6[Ta10O28]·6H2O revealed that the anion has the decametalate structure and is isostructural with the decavanadate and decaniobate anions [a = 15.8517(8) Å, b = 19.364(1) Å, c = 21.935(1) Å, ß = 93.638(1)°, V = 6719.4(6) Å(3), Z = 2, and space group P2(1)/n at 292(2) K].

The Effects of Childhood Health on Adult Health and SES in China.

In this paper, we model the associations of childhood health on adult health and socio-economic status outcomes in China using a new sample of middle aged and older Chinese respondents. Modeled after the American Health and Retirement Survey (HRS), the CHARLS Pilot survey respondents are 45 years and older in two quite distinct provinces-Zhejiang, a high growth industrialized province on the East Coast and Gansu, a largely agricultural and poor province in the West. Childhood health in CHARLS relies on two measures that proxy for different dimensions of health during the childhood years. The first is a retrospective self-evaluation using a standard five-point scale (excellent, very good, good, fair, or poor) of general state of one's health when one was less than 16 years old. The second is adult height believed to be a good measure of levels of nutrition during early childhood and the prenatal period. We relate both these childhood health measures to adult health and SES outcomes during the adult years. We find strong associations of childhood health on adult health outcomes particularly among Chinese women and strong associations with adult BMI particularly for Chinese men.

Vps26A and Vps26B subunits define distinct retromer complexes.

The trimeric Vps29-Vps35-Vps26 sub-complex of retromer mediates retrograde transport of transmembrane proteins from endosomes to the trans-Golgi network. Our group has recently identified a Vps26 paralogue, Vps26B, which is able to suppress the expression of Vps26A when exogenously expressed in mammalian cells and defines a distinct retromer complex (Vps26B-retromer) in vivo and in vitro. In this study, we use HEK293 cells stably expressing either Vps26A-myc or Vps26B-myc to address the role of retromer cargo transport and subcellular localization of the two core retromer complexes as defined by the two mammalian-specific Vps26 paralogues. Vps26B-retromer, like Vps26A-retromer, associates with TBC1D5 and GOLPH3. In contrast, no interaction between Vps26B-retromer and cation-independent mannose 6-phosphate receptor (CI-M6PR) was detected, leading to a degradation of this receptor and an increase in cathepsin D secretion. Colocalization of Vps26 paralogues with different endosomally located Rab proteins shows prolonged association of Vps26B-retromer with maturing endosomes relative to Vps26A-retromer. Interestingly, the cycling of CI-M6PR is restored upon deletion of the variable Vps26B C-terminal region indicating that this region is directly responsible for the differential function of the two paralogues. In summary, we show that the two distinct retromer complexes defined by different Vps26 paralogues are not functionally equivalent and that the Vps26B C-terminal region can control cargo selection of the Vps26B-retromer.

Pomegranate flower ameliorates fatty liver in an animal model of type 2 diabetes and obesity.

AIMS OF THE STUDY: Fatty liver is the most common cause of abnormal liver function tests. We investigated the effect and its underlying mechanism of pomegranate flower (PGF), a traditional antidiabetic medicine, on fatty liver. MATERIALS AND METHODS: At the endpoint of treatment of male Zucker diabetic fatty (ZDF) rats with PGF extract (500 mg/kg, p.o. x 6 weeks), liver weight index, hepatic lipid contents (enzymatic colorimetric methods) and droplet accumulation (Oil Red O staining) were determined. Gene profiles (RT-PCR) were analyzed in the liver of ZDF rats and in human liver-derived HepG2 cell line. RESULTS: PGF-treated ZDF rats showed reduced ratio of liver weight to tibia length, hepatic triglyceride contents and lipid droplets. These effects were accompanied by enhanced hepatic gene expression of peroxisome proliferator-activated receptor (PPAR)-alpha, carnitine palmitoyltransferase-1 and acyl-CoA oxidase (ACO), and reduced stearoyl-CoA desaturase-1. In contrast, PGF showed minimal effects on expression of genes responsible for synthesis, hydrolysis or uptake of fatty acid and triglycerides. PGF treatment also increased PPAR-alpha and ACO mRNA levels in HepG2 cells. CONCLUSION: Our findings suggest that this Unani medicine ameliorates diabetes and obesity-associated fatty liver, at least in part, by activating hepatic expression of genes responsible for fatty acid oxidation.

Increased renal collagen cross-linking and lipid accumulation in nephropathy of Zucker diabetic fatty rats.

BACKGROUND: Zucker diabetic fatty (ZDF) rat is a genetic model of type 2 diabetes and obesity. The mechanism underlying nephropathy in ZDF rats, however, remains unclear. METHODS: ZDF rats were compared to age-matched Zucker lean (ZL) rats. Physiological and blood biochemical parameters, renal glomerular cross-sectional area (hematoxylin-eosin staining), fibrosis (van Giesen staining), collagen composition (Sircol Collagen Assay), lipids (enzymatic method) and mRNA expression (RT-PCR) were determined. RESULTS: ZDF rats showed an increase in renal-insoluble collagen content and the ratio of renal-insoluble to salt-soluble collagen (2- and 1.5-fold of the control animals). There were increases in renal glomerulosclerosis and interstitial fibrosis in ZDF rats (increased to 2-fold) in the glomerular mesangium and tubulointerstitium, and increased glomerular area. Renal triglyceride accumulated to greater than 2-fold of those levels in ZL rats. These changes were accompanied by hypoalbuminemia, and elevated plasma blood urea nitrogen and uric acid levels. Gene profiling showed increased expression of transcripts encoding the glomerulosclerotic mediator collagens I and IV, plasminogen activator inhibitor-1, transforming growth factor-beta1, and angiotensin II type 1 receptor in ZDF rat kidney. Moreover, renal expression of mRNAs encoding sterol regulatory element-binding protein-1, a nuclear transcription factor that activates genes involved in fatty acid synthesis, and acetyl-CoA carboxylase, a key enzyme that mediates fatty acid synthesis, was increased in ZDF rats. CONCLUSIONS: Our findings suggest that dysregulated gene expression may result in increased renal collagen cross-linking and lipid accumulation, that may be associated with development of nephropathy in the animal model of type 2 diabetes and obesity.

A novel deletion in TNNI2 causes distal arthrogryposis in a large Chinese family with marked variability of expression.

Distal arthrogryposis (DA) is composed of a group of clinically and genetically heterogeneous disorders, characterized by multiple congenital contractures of the limbs. Point mutations in three genes encoding contractile fast-twitch myofibers, TPM2, TNNI2 and TNNT3, were recently identified in DA type 1 (DA1; MIM 108120) and DA type 2B (DA2B; MIM 601680). We have described a large Chinese DA family in which different individuals had phenotypes similar to DA1 or DA2B. To map the disease locus in this family, two-point linkage analysis was first performed using microsatellite markers selected from the genomic regions close to the TPM2, TNNI2/TNNT3 and TNNC2 genes. A positive LOD score of 3.61 at theta = 0 was obtained with the marker close to the TNNI2/TNNT3 genes, corresponding to the genetic mapping site of DA2B. Direct sequencing of the PCR-amplified DNA fragment spanning exon 8 of the TNNI2 gene showed a heterozygous deletion, c.523_525delAAG (p.K175del), in the proband. This novel mutation was confirmed to cosegregate with the DA phenotype in affected individuals but not detected in all unaffected individuals of the family and not in 50 healthy controls. In summary, we have found a novel TNNI2 mutation in a Chinese family with DA2B. Our work represents the first report on the link between TNNI2 and the DA phenotype in Chinese.

The subcellular fractionation properties and function of insulin receptor substrate-1 (IRS-1) are independent of cytoskeletal integrity.

Efficient insulin action requires spatial and temporal coordination of signaling cascades. The prototypical insulin receptor substrate, IRS-1 plays a central role in insulin signaling. By subcellular fractionation IRS-1 is enriched in a particulate fraction, termed the high speed pellet (HSP), and its redistribution from this fraction is associated with signal attenuation and insulin resistance. Anecdotal evidence suggests the cytoskeleton may underpin the localization of IRS-1 to the HSP. In the present study we have taken a systematic approach to examine whether the cytoskeleton contributes to the subcellular fractionation properties and function of IRS-1. By standard microscopy or immunoprecipitation we were unable to detect evidence to support a specific interaction between IRS-1 and the major cytoskeletal components actin (microfilaments), vimentin (intermediate filaments), and tubulin (microtubules) in 3T3-L1 adipocytes or in CHO.IR.IRS-1 cells. Pharmacological disruption of microfilaments and microtubules, individually or in combination, was without effect on the subcellular distribution of IRS-1 or insulin-stimulated tyrosine phosphorylation in either cell type. Phosphorylation of Akt was modestly reduced (20-35%) in 3T3-L1 adipocytes but not in CHO.IR.IRS-1 cells. In cells lacking intermediate filaments (Vim(-/-)) IRS-1 expression, distribution and insulin-stimulated phosphorylation appeared normal. Even after depolymerisation of microfilaments and microtubules, insulin-stimulated phosphorylation of IRS-1 and Akt were maintained in Vim(-/-) cells. Taken together these data indicate that the characteristic subcellular fractionation properties and function of IRS-1 are unlikely to be mediated by cytoskeletal networks and that proximal insulin signaling does not require an intact cytoskeleton.