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15. Curtiss R III: Antigen delivery systems: Development of live recombinant attenuated bacterial antigen and DNA vaccine delivery vector vaccines. In Mucosal Immunology. Edited by: Mestecky J, Lamm ME, Strober W, Bienenstock J, McGhee JR, Mayer L. San Diego: Elsevier Academic Press; 2005:1009–1037.CrossRef 16. Luo Y, Zhou H, Mizutani M, Mizutani N, Reisfeld RA, Xiang R: Transcription factor Fos-related antigen

1 is an effective target for a breast cancer vaccine. Proc Natl Acad Sci USA 2003,100(15):8850–8855.PubMedCrossRef 17. Zhang X, Kong W, Ashraf S, Curtiss R III: A one-plasmid system to generate influenza virus in cultured chicken cells for potential use in influenza vaccine. J Seliciclib order Virol 2009,83(18):9296–9303.PubMedCrossRef 18. Li Y, Wang S, Scarpellini G, Gunn B, Xin W, Wanda SY, Roland KL, Curtiss R III: Evaluation of new generation Salmonella enterica serovar Typhimurium vaccines with regulated delayed attenuation to induce immune responses against PspA. Proc Natl Acad Sci USA 2009,106(2):593–598.PubMedCrossRef 19. Curtiss R III, Wanda SY, Gunn BM, Zhang X, Tinge SA, Ananthnarayan V, Mo H, Wang S, Tangeritin Kong W: Salmonella enterica serovar Typhimurium strains with regulated delayed attenuation in vivo . Infect Immun 2009,77(3):1071–1082.PubMedCrossRef 20. Konjufca V, Jenkins M, Wang S, Juarez-Rodriguez

MD, Curtiss R III: Immunogenicity of recombinant attenuated Salmonella enterica serovar Typhimurium vaccine strains carrying a gene that encodes Eimeria tenella antigen SO7. Infect Immun 2008,76(12):5745–5753.PubMedCrossRef 21. Xin W, Wanda SY, Li Y, Wang S, Mo H, Curtiss R III: Analysis of type II secretion of recombinant pneumococcal PspA and PspC in a Salmonella enterica serovar Typhimurium vaccine with regulated delayed antigen synthesis. Infect Immun 2008,76(7):3241–3254.PubMedCrossRef 22. Wang S, Li Y, Scarpellini G, Kong W, Shi H, Baek CH, Gunn B, Wanda SY, Roland KL, Zhang X, et al.: Salmonella vaccine vectors displaying delayed antigen synthesis in vivo to enhance immunogenicity. Infect Immun 2010,78(9):3969–3980.PubMedCrossRef 23. Kong W, Wanda SY, Zhang X, Bollen W, Tinge SA, Roland KL, Curtiss R III: Regulated programmed lysis of recombinant Salmonella in host tissues to release protective antigens and confer biological containment. Proc Natl Acad Sci USA 2008,105(27):9361–9366.PubMedCrossRef 24.

After 48 h of transfection, fluorescence of cells was observed by a fluorescence microscope. Then, cells were seeded

for FCM and immunofluorescence assay. Supernatant was collected to test the inflammatory cytokines secreted by the cells. Table 2 sequences of siRNA against TLR4 Name of siRNA TLR4 sequences(5′-3′) Site position TLR4A a a c t t g t a t t c a a g g t c t g g c 1023-1044 TLR4B a a g g c t t a c t t t c a c t t c c a a 1374-1395 TLR4C a a c t c c c t c c a g g t t c t t g a t 1921-1942 MTT assay Cells were seeded into 96-well culture plates (6×103/well, 5 wells repeated), allowed to adhere overnight, and then transfections were performed according to the manufacturer’s instructions. After 48 h, the transfected cells were collected (0 h) or allowed to continue in RAD001 concentration culture for 24 h, 48 h, or 72 h. At the end of each treatment, selleck products cells were incubated with 5 mg/mL MTT (Sigma Chemical

Co., MO, USA) for 4 h and then mixed with dimethyl sulfoxide after the supernatant was removed. The dye absorption (A) was quantitated using an automatic microplate spectrophotometer (340 st; Anthos Zenyth, Salzburg, Austria) at 490 nm. Human inflammatory cytokine assay IL-6 and IL-8 presence in the supernatant of transfected cells were detected according to the instruction of human inflammatory cytokine kit (BD™ Cytometric Bead Array (CBA)). FACScan flow cytometer (BD) was used to analyze samples. Statistical Analysis GraphPad Prism software (CA, USA) was used to perform statistical comparisons between different values. Data were expressed as the means ± standard deviation (SD) with n = 3. Statistical significances were determined by Student’s t-test and ANOVA, differences were considered significant at a P value of less

than 0.05. Results Dynein Expression of TLRs in human breast cancer cell line MDA-MB-231 As TLRs have been identified in some tumor cells, we sought to detect if they were expressed in the human breast cancer cell line MDA-MB-231. Qualitative RT-PCR analysis revealed that MDA-MB-231 expressed mRNA of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9 and TLR10 (Figure 1A). Real-time PCR analysis revealed the relative expressions of each TLR examined. The expression of TLR3 was normalized to 1.0, as it was expressed the most weakly. TLR4 was 5-fold higher than TLR3, while other TLRs were expressed between 1- and 4-fold higher than TLR3 (Figure 1B). By FCM detection, we were able to examine the different protein expression levels of the TLRs, TLR4, TLR6, TLR7 and TLR5 were expressed moderately; the other TLRs were expressed weakly or unexpressed. Again, TLR4 protein level was the highest out of TLR1-TLR10 (Figure 1C). Collectively, these results demonstrated that MDA-MB-231 expressed all the TLRs examined (TLR1-TLR10) and TLR4 was expressed highest. TLR4 was strategically selected to investigate its function on the growth and progression of MDA-MB-231 in subsequent studies.

004581387 0.008668512 0.53 2 0.011048543 0.015517070 0.71 3 0.009226505 0.013696964 0.67 4 0.011280697 0.015843117 0.71 5 0.010525262 0.014578640 0.72 6 0.006258358 0.016064279 0.39 7 0.003569654 0.031034140 0.12 8 0.003721242 0.035402621 0.10 9 0.002008035 0.020617311 0.10 10 0.018073253 0.028955877 0.63 11 0.002800694 0.015303442 0.18 12 0.010096506 0.017701311 0.57 13 0.005083367 0.019505165 0.26 miR-320c suppresses bladder cancer cell viability, inhibits clone formation

and triggers G1-phase arrest In order to understand the potential mechanisms of miR-320c in tumor suppressing, the bladder cancer cell lines were transfected with miR-320c to evaluate the effect of over-expression Z-VAD-FMK mw via cell viability assay. As a result, miR-320c illustrated a significant inhibitory effect on bladder cancer cell viability in a dose-dependent manner (Figure 2A). After 48 h transfection, miR-320c (50nM) could reduce cell viability in

both UM-UC-3 and T24 cell by 35% and 49%, respectively. Furthermore, miR-320c potently inhibited the colony forming ability in both cell lines. Compared with cell lines transfected with NC, the colony formation rate decreased drastically GSK-3 inhibitor in those transfected with miR-320c (Figure 2B). Figure 2 Over-expression of miR-320c suppresses bladder cancer cell proliferation and motility. (A) Cell viability assay. The relative cell viability was lower in the miR-320c treated groups (cell viability of 0nM was regarded as 1.0), respectively. (B) Colony formation assay (representative wells were presented). The colony formation rate was lower in miR-320c treated groups. (C) miR-320c impaired the motility of both cell lines (representative

migration and invasion results at × 200 were presented). (D) Cell cycle distribution in bladder cancer cell lines. Over-expression of miR-320c induced G1-phase arrest in both cell lines (representative histograms were presented) (*P < 0.05). Additionally, in order to GNAT2 better clarify the underlying mechanisms for miR-320c inhibiting cancer cell proliferation, we transfected the cells with 50nM miR-320c 48 h before assessing the impact of miR-320c on cell cycle distribution via flow cytometry. As a result, we observed a significant increase in the percentage of cells in the G1/G0 phase and a decrease in the percentage of cells in the S and G2/M phase in miR-320c-overexpressing cells (Figure 2D). These results suggested that miR-320c could lead to G1-phase arrest. miR-320c impairs UM-UC-3 and T24 cell motility To further elucidate the function of miR-320c, we investigated the potential effect of miR-320c on UM-UC-3 and T24 cell motility. As illustrated by the transwell assay, over-expression of miR-320c decreased the migration and invasion of cancer cells compared with NC (Figure 2C). Therefore, miR-320c negatively regulated the motility of UM-UC-3 and T24 cells.

Additionally, our patient was on hemorrhagic diathesis with the oral anticoagulation Rucaparib cell line therapy for

atrial fibrillation, and attended with suspicious disseminated intravascular coagulation due to massive hemorrhage. But it wcxxas expected that the major vascular leakage was only in the hepatic arterial branch without any bowel perforation on the contrast-enhanced CT, so we performed interventional procedure. NBCA was the most appropriate embolic agent of TAE for our case with hemorrhagic diathesis, because it does not depend on the coagulation process for its therapeutic effect [8]. There are some reports of ACS treated with TAE [9]. However, combination treatment selleck chemicals of TAE with NBCA and percutaneous catheter drainage (PCD) for ACS has not been reported (Table  1). We suggest that initial hemostasis by transcatheter arterial embolization is a safe, effective treatment method for abdominal compartment syndrome with active arterial bleeding in a patient undergoing anticoagulation. Table 1 The characteristics

of the reported cases of abdominal compartment syndrome treated with transcatheter arterial embolization Author N Clinical presentation Embolized artery Embolic material Subsequent treatment Letoublon [9] 14 Blunt hepatic trauma Hepatic artery NS Decompressive laparotomy or laparoscopy Won [10] 1 Retroperitoneal hemorrhage Internal iliac artery Gelatin sponge, coil, lipiodol Decompressive laparotomy Pena [11] 1 Splenomegaly Splenic artery PVA Nothing Monnin [12] 7 Blunt hepatic trauma Hepatic artery Gelatin sponge, coil Decompressive laparotomy         Trisacryl gelatin microsphere   Hagiwara [13] 1 Pelvic flactures Bortezomib nmr Super gluteal artery Gelatin sponge Repeat TAE, decompressive laparotomy

Isokangas [14] 5 Retroperitoneal hemorrhage Lumbar artery (N = 4) Gelatin sponge, PVA, coil Surgical decompreesion (N = 4)       Medial rectal artery (N = 1)   US guided drainage (N = 1) Tokue (present) 1 Blunt hepatic trauma Hepatic artery NBCA, lipiodol US guided drainage N: number of patients, NS: not shown, PVA: polyvinyl alcohol, NBCA: N-Butyl Cyanoacylate, US: ultrasonography. The decompression is simultaneously essential to hemostasis for the treatment of primary ACS. There are some randomized controlled trials for ACS (Table  2) [31]. However, there have been no randomized controlled trials about which is better, PCD or decompressive laparotomy. PCD is easy and minimal invasive procedure compared with surgical decompression, and allows us to measure IAP. But it is not appropriate to perform catheter drainage for the patients with widespread peritonitis or bowel injury.

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The applicability of the swine model for human liver injury has been well described in the literature. This model, however, is not without its limitations. The compression of the portal inflow during creation of the liver laceration minimized initial blood losses. In the clinical setting, uncompensated hypovolemic shock may result in the ‘bloody vicious cycle’

of hypothermia, acidosis, and coagulopathy. Obtaining check details hemostasis from bleeding viscera in the face of these physiologic derangements can be quite challenging. In this regard, the model used for this experiment was artificial given that the pig was well compensated hemodynamically, with functioning coagulation cascades. However, given the mechanism of action of the VAC device, the authors contend that L-VAC placement may be the ideal therapy for control of hemorrhage

in such cases. Consideration is being given to repeating this experiment in animals that are hypothermic and coagulopathic. Future areas of investigation should be directed toward comparing this innovative method to Linsitinib research buy well-established therapies such as packing, mesh wrapping, and application of hemostatic agents. In summary, these data demonstrate the feasibility and utility of a perihepatic negative pressure device for the treatment of hemorrhage from severe liver injury in the porcine model. This method is potentially applicable in the clinical setting and may afford advantages over traditional damage control procedures such as perihepatic packing. Financial disclosure This study was funded in part by funds

from the Kansas University Medical Center, and the Wesley Medical Center Trauma Research Fund. Institutional animal use and care committee approval This study was approved for implementatin by the IACUC of the Kansas University Medical center. References 1. Pachter HL, Liang HG, Hofstetter SR: Liver and biliary tract trauma. In Trauma. 3rd edition. Edited by: Feliciano DV, Moore EE, Mattox KL. Stamford, CT: Appleton & Lange; 1996:487. 2. Richardson JD, Franklin GA, Lukan JK, Carrillo EH, Spain DA, Miller FB, Wilson MA, Polk HC Jr, Flint LM: Evolution in the management of hepatic trauma: a 25-year perspective. Ann Surg 2000, 232:324–330.CrossRef 3. Malhotra AK, Fabian TC, Croce MA, Gavin TJ, Kudsk KA, Minard G, Pritchard FE: Blunt hepatic injury: a paradigm shift from operative to nonoperative management in the 1990s. Ann Surg 2000, 231:804–813.PubMedCrossRef Farnesyltransferase 4. Moore EE, Shackford SR, Pachter HL, McAninch JW, Browner BD, Champion HR, Flint LM, Gennarelli TA, Malangoni MA, Ramenofsky ML, Trafton PG: Organ injury scaling: spleen, liver, and kidney. J Trauma 1989, 29:1664–1666.PubMedCrossRef 5. Aaron S, Fulton RL, Mays ET: Selective ligation of the hepatic artery for trauma of the liver. Surg Gynecol Obstet 1975, 141:187–189.PubMed 6. Stone HH, Lamb JM: Use of pedicled omentum as an autogenous pack for control of hemorrhage in major injuries of the liver. Surg Gynecol Obstet 1975, 141:92–94.PubMed 7.

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G: Adhesion and inhibition assay of Mycoplasma genitalium and M. pneumoniae by immunofluorescence microscopy. J Med Microbiol 2002,51(5):361–373.PubMed 15. Chaudhry R, Varshney AK, Malhotra P: Adhesion proteins of Mycoplasma pneumoniae . Front Biosci 2007, 12:690–699.PubMedCrossRef 16. Krivan HC, Olson LD, Barile MF, Ginsburg V, Roberts DD: Adhesion of Mycoplasma pneumoniae to sulfated glycolipids and inhibition by dextran sulfate. J Biol Chem 1989,264(16):9283–9288.PubMed 17. Krause DC, Leith DK, Wilson RM, Baseman JB: Identification of Mycoplasma pneumoniae proteins associated with hemadsorption and virulence. Infect Immun 1982,35(3):809–817.PubMedCentralPubMed 18. Krause DC: Mycoplasma pneumoniae cytadherence: unravelling

the tie that binds. Mol Microbiol 1996,20(2):247–253.PubMedCrossRef 19. Seto S, Kenri T, Tomiyama T, Miyata M: Involvement of P1 adhesin in gliding motility of Mycoplasma Selleckchem PD98059 pneumoniae as revealed by the inhibitory effects of antibody under optimized gliding conditions. J Bacteriol 2005,187(5):1875–1877.PubMedCentralPubMedCrossRef 20. Tabassum I, Chaudhry R, Chourasia BK, Malhotra P: Identification of N-terminal 27 kDa fragment of Mycoplasma pneumoniae P116 protein as specific immunogen in M. pneumoniae infections. BMC Infect Dis 2010,

10:350.PubMedCentralPubMedCrossRef 21. Chaudhry R, Nisar N, Hora B, Chirasani SR, Malhotra P: this website Expression and immunological characterization of the carboxy-terminal region of the P1 adhesin protein of Mycoplasma pneumoniae . J Clin Microbiol 2005,43(1):321–325.PubMedCentralPubMedCrossRef 22. Jacobs E, Fuchte K, Bredt W: Isolation of the adherence protein of Mycoplasma pneumoniae by fractionated solubilization and size exclusion chromatography. Biol Chem Hoppe Seyler 1988,369(12):1295–1299.PubMedCrossRef 23. Dallo SF, Su CJ, Horton JR, Baseman JB: Identification of P1 gene domain containing epitope(s) mediating Mycoplasma pneumoniae cytoadherence. J Exp Med 1988,167(2):718–723.PubMedCrossRef 24. Opitz O, Jacobs E: Adherence epitopes of Mycoplasma genitalium adhesin. J Gen Microbiol 1992,138(9):1785–1790.PubMedCrossRef 25. Jacobs E, Pilatschek A, Gerstenecker B, Oberle K, Bredt W: Immunodominant epitopes of the adhesin of Mycoplasma pneumoniae . J Clin Microbiol 1990,28(6):1194–1197.PubMedCentralPubMed 26.

[35] India, PF-02341066 solubility dmso Kashmir valley, all year round Indian M, mean 29 years (n = 64) 38 ± 30, 41% < 25 Lower exposure to sunlight, female gender Indian F, mean 27 years (n = 28) 14 ± 11, 96% < 25 Gulvady et al. [44] India, Mumbai Indian M, 40–68 years, senior executives

(indoor workers; n = 86) 28% < 19 Earlier start of the workday Vupputuri et al. [43] India, Delhi (28° N) Asian Indian M, mean 43 years (for both men and women), urban, middle income, mostly working indoors (n = 51) 27 ± 17 – Asian Indian F, mean 43 years (for both men and women), urban, middle income, mostly housewives (n = 54) 22 ± 12 Harinarayan [65] India, Tirupati (13° N), all year round Indian F, mean 54 years, postmenopausal (n = 164) 37 ± 18, 30% < 25 Higher EX 527 research buy dietary calcium intake, higher dietary phytate intake, higher phytate to calcium ratio Harinarayan et al. [21] India, around Tirupati (13° N), winter to summer (Jan–Jul) Indian, mean 44 years, rural (n = 191) 53 ± 06, 03% < 25 Urban subject, lower dietary calcium intake, higher phytate to calcium ratio Indian, mean 46 years, urban (n = 125) 34 ± 07, 35% < 25 Goswami et al. [18] India, Dehli (28° N), in winter or

summer Indian M, mean 25 years, soldiers, winter new (n = 31) 47 ± 12 Less exposure to sunlight, more skin pigmentation, winter season Indian M (58%)+F, mean 23 years, physicians and nurses, winter (n = 19) 08 ± 03 Indian M (67%)+F, mean 43 years, depigmented persons, winter (n = 15) 18 ± 11 Indian M (58%)+F, mean 24 years, physicians and nurses, summer (n = 19) 18 ± 08 Pregnant women Sahu et al. [36] India, Barabanki

district, 32 km from Lucknow (27°), all year round Indian, rural, mean 27 years (n = 139) 38 ± 20, 32% < 25 Lower summer sun exposure, measurement in winter Farrant et al. [66] India, Mysore (South India) at the 30th week of pregnancy Indian, mean 24 years (n = 559) Median 38, 31% < 28 nmol/l Taking calcium and vitamin D at recruitment, measurement in Mar–Aug Bhalala et al. [45] Western India, at the 37th week of pregnancy, all year round Indian, 20–35 years, middle income group (n = 42) 57 ± 27 Lower serum 25(OH)D in mother → lower serum 25(OH)D in cord blood Cord blood (n = 42) 48 ± 24 Sachan et al. [46] India, Lucknow (27° N), before labor, autumn Indian, total group (n = 207) 43% < 25 – Indian, urban (n = 140) 35 ± 24 Indian, rural (n = 67) 35 ± 22 Goswami et al. [18] India, Dehli (28° N), in summer Indian, mean 23 years, poor socioeconomic class (n = 29) 22 ± 11 – Children Sahu et al.

After incubation, 100 μl DMSO were added to each well, and the culture plate was vortexed for 2-3 min to fully dissolve the crystallization. Finally, the absorbance at 562 nm was measured using microplate reader. FITC- Gelatin degradation assay FITC-gelatin degradation assay was performed as the manufacture’s procedure (Invitrogen). In brief, coverslips (18-mm diameter) were coated with 50ug/ml poly-L-lysine for 20 min at room temperature,

washed with PBS, fixed with 0.5% glutaraldehyde for 15 min and washed with PBS for 3 times. After washing, the coverslips were inverted on a drop of 0.2% FITC conjugated gelatin in PBS containing 2% sucrose, incubated for 10 min at room temperature, washed with PBS for 3 times, quenched with sodium borohydride (5 mg/ml) for 3 min and finally incubated in 2 ml of complete medium for 2 h. Cells (2 × 105 each well) were plated in FITC Selleck Lumacaftor gelatin-coated coverslips, incubated at 37°C for 12 hr. The ECM degradation status was evaluated and photographed by inverted fluorescent microscope. Gelatin zymography The Conditioned medium was Adriamycin in vivo collected and concentrated for 2-fold by centrifugal concentrator. Equal amounts of protein were loaded and separated by 10% polyacrylamide gel containing

1 g/L gelatin. The gels were re-natured in 2.5% Triton-X-100 with gentle agitation for 30 min at room temperature. The gel was pretreated by developing buffer (5 mM CaCl2, 50 mM Tris, and 0.2 mM NaCl, 0.02% Brij35 (pH 7.5)) for 30 min at room temperature, then developed in developing buffer overnight at 37°C, stained with Coomassie Brilliant Blue R-250 for 30 minutes and destained with destaining solution. The protease activity was analyzed by gel imaging and analysis system. Statistical analysis The results were represented as ± SE. Difference between two experimental groups was evaluated by the students’t test and differences among groups were analyzed using One-Way ANOVA. P < 0.05 was considered to be

statistically significant. Acknowledgement check details This article is financially supported by the Natural Science Foundation of China (81172048) and the Science and Technology Development Project of Liaoning province of China(2008225010–17). References 1. EI-Serag HB, Rudolph KL: Hepatocellular carcinoma: epidemiology and molecular carcinogenesis. Gastroenterology 2007, 132:2557–2576.CrossRef 2. Blagden SP, Willis AE: The biological and therapeutic relevance of mRNA translation in cancer. Nature Review Clinical Oncology 2011, 8:280–291.CrossRef 3. Pfaffenbach KT, Lee AS: The critical role of GRP78 in physiologic and pathologic stress. Curr Opin Cell Biol 2011, 23:150–156.PubMedCrossRef 4. Gonzalez-Gronow M, Selim MA, Papalas J, Pizzo SV: GRP78: a multifunctional receptor on the cell surface. Antioxid Redox signal 2009, 11:2299–2306.PubMedCrossRef 5.

For each activity, the frequency, duration

in minutes, and MET score were multiplied and then divided by 14 days (i.e., (frequency × duration × MET)/14). The minutes spent per activity per day were summed to a total physical activity score (minutes/day × MET). For example, a participant who walks outside for 60 min four times per 2 weeks (4 × 60 × 3.5/14 = 60) and does light HDAC assay household work for 30 min per day (14 × 30 × 2.5/14 = 75) has a physical activity score of 135 min/day × MET. Potential effect modifiers Physical functioning was measured by physical performance and functional limitations. Physical performance was measured using the chair stands test (time needed to stand up from a chair and sit down for five times), the walk test (time needed to walk 3 m, turn 180°, and walk back), and the tandem stand (the participant stands unsupported with one foot behind the other (heel against toe) up to 10 s) [23, 29]. In order to calculate a total physical performance score, the time needed for the chair stands and walk test were categorized into quartiles (1 = slowest, 4 = fastest). For the tandem stand, 2

points were scored when able to hold for 3 to 9 s, and 4 points for 10 s. For each test, the score of 0 was assigned when the participant was unable to complete the test. The three scores were summed (range 0–12), a score of 12 representing optimal physical performance. Functional limitations were assessed using a validated questionnaire about the degree of difficulty with climbing stairs, walking 5 min outdoors without resting, getting Selleckchem 5-Fluoracil Tangeritin up and sitting down

in a chair, dressing and undressing oneself, cutting one’s toenails, and using own or public transportation [30]. The scores on these six items were dichotomized (0 = no difficulty, 1 = at least some difficulty) and summed (range 0–6). A score of 6 indicates difficulties with all six activities. We dichotomized both measures, because, in case of a significant interaction with physical activity, further analyses would have to be stratified into low and high physical functioning, and stratification for more than two groups would have severely decreased the power to detect a significant association between physical activity and fall risk. Physical performance was dichotomized using the median score of 7 as the cut-off value (0–7 vs 8–12). Functional limitations were dichotomized using the median score of 1 as a cut-off value (0 vs ≥1 limitations). Confounders BMI (Body Mass Index) was calculated as weight (kilograms)/height (square meter). The number of chronic diseases was assessed using self-reports on chronic diseases, which included chronic nonspecific lung diseases, cardiac diseases, vascular diseases, stroke, diabetes mellitus, malignant neoplasms, and joint disorders (i.e., osteoarthritis and rheumatoid arthritis; range 0–7) [31]. Medication use was assessed by recording the names of the medications directly from the containers.