Infect Immun 1997, 65:1172–1180.PubMed

16. Tannaes T, Bukholm IK, Bukholm G: High relative content of lysophospholipids of Helicobacter pylori RG7420 ic50 mediates increased risk for ulcer disease. FEMS Immunol Med Microbiol 2005, 44:17–23.PubMedCrossRef 17. Marshall BJ, Warren JR: Unidentified curved bacilli in the stomach of patients with gastritis and peptic ulceration. Lancet 1984, 1:1311–1315.PubMedCrossRef 18. You YH, Song YY, Meng FL, He LH, Zhang MJ, Yan XM, et al.: Time-series gene expression profiles in AGS cells stimulated with Helicobacter pylori. EVP4593 chemical structure World J Gastroenterol 2010, 16:1385–1396.PubMedCrossRef 19. Wang SY, Shen XY, Wu CY, Pan F, Shen YY, Sheng HH, et al.: Analysis of whole genomic expression profiles of Helicobacter pylori related chronic atrophic gastritis with IL-1B-31CC/-511TT genotypes. J Dig Dis 2009, 10:99–106.PubMedCrossRef 20. Shibata W, Hirata Y, Yoshida H, Otsuka M, Hoshida Y, Ogura K, et al.: NF-kappaB and ERK-signaling pathways contribute to the gene expression induced by cag PAI-positive-Helicobacter pylori infection.

World J Gastroenterol 2005, 11:6134–6143.PubMed 21. Sepulveda AR, Tao H, Carloni E, Sepulveda J, Graham Dorsomorphin in vitro DY, Peterson LE: Screening of gene expression profiles in gastric epithelial cells induced by Helicobacter pylori using microarray analysis. Aliment Pharmacol Ther 2002,16(Suppl 2):145–157.PubMedCrossRef 22. Nagasako T, Sugiyama T, Mizushima T, Miura Y, Kato M, Asaka M: Up-regulated Smad5 mediates apoptosis of gastric epithelial cells induced by Helicobacter pylori infection. J Biol Chem 2003, 278:4821–4825.PubMedCrossRef 23. Maeda S, Otsuka M, Hirata Y, Mitsuno

Y, Yoshida H, Shiratori Y, et al.: cDNA PR171 microarray analysis of Helicobacter pylori-mediated alteration of gene expression in gastric cancer cells. Biochem Biophys Res Commun 2001, 284:443–449.PubMedCrossRef 24. Liu YJ, Yan PS, Li J, Jia JF: Expression and significance of CD44s, CD44v6, and nm23 mRNA in human cancer. World J Gastroenterol 2005, 11:6601–6606.PubMed 25. Lim JW, Kim H, Kim KH: Cell adhesion-related gene expression by Helicobacter pylori in gastric epithelial AGS cells. Int J Biochem Cell Biol 2003, 35:1284–1296.PubMedCrossRef 26. Kim N, Park WY, Kim JM, Park YS, Lee DH, Park JH, et al.: Analysis of gene expression profile of AGS cells stimulated by Helicobacter pylori adhesion. Gut Liver 2007, 1:40–48.PubMedCrossRef 27. Han YH, Liu WZ, Shi YZ, Lu LQ, Xiao SD, Zhang QH: Gene expression profile of Helicobacter pylori in response to growth temperature variation. J Microbiol 2009, 47:455–465.PubMedCrossRef 28. Ding SZ, Torok AM, Smith MF Jr, Goldberg JB: Toll-like receptor 2-mediated gene expression in epithelial cells during Helicobacter pylori infection. Helicobacter 2005, 10:193–204.PubMedCrossRef 29. Guillemin K, Salama NR, Tompkins LS, Falkow S: Cag pathogenicity island-specific responses of gastric epithelial cells to Helicobacter pylori infection.

5 ± 3.1 51.3 ± 3.0 5.6 ± 0.7 2.6 ± 2.3 HL1 with AtMinD 50 μM 8.7 ± 0.8 87.4 ± 2.5 3.9 ± 1.8 0 HL1 with EcMinD 20 μM 0 0 0 100 RC1 with AtMinD 50 μM 31.5 ± 1.5 48.8 ± 1.3 16 ± 4.4 5.5 ± 2.8 HL1 with AtMinD-GFP 50 μM 12.5 ± 2.4 78.6 ± 2.5 7.6 ± 1.1 1.3 ± 0.3

HL1 with GFP-AtMinD 50 μM 5.2 ± 1.5 91.5 ± 2.7 3.3 ± 1.3 0 Shown above are the means ± S.D. obtained from 3 independent repeats. The number of the cells measured in each repeat is between 150 and 200. Table 2 Analysis of the cell division phenotype Genotype Cells Septa Polar % Polar Phenotype DH5α 867 229 6 3 WT HL1 991 216 119 55 Min- HL1(Plac::EcMinDE) 974 232 3 1 WT HL1(Plac::AtMinD) 863 161 11 6 WT HL1(Plac::gfp-AtMinD) 1081 219 10 5 WT HL1(Plac::AtMinD-gfp) 943 137 17 12 WT like Shown above is the division phenotype analysis of E. coli cells with different genotypes. EcMinDE was induced with 20 μM IPTG, AtMinD PLX3397 datasheet and its GFP fusion proteins were induced with 50 μM IPTG. Cells: the total number of cell examined; Septa: the total number of septa counted; Polar: the number

of septa which were misplaced at or near a cell pole; % Polar: the percentage of septa which were misplaced at or near a cell pole. Min-, minicell phenotype. P005091 solubility dmso WT, most of the cells have a normal size and no cell or only a small part of the cells are minicells or long filaments. Figure 1 The phenotype of E. coli cells. (A) Wildtype, DH5α. (B) HL1 mutant (ΔMinDE). (C) HL1 mutant (ΔMinDE) complemented by pM1113-MinDE at 20 μM IPTG. (D) HL1 mutant (ΔMinDE) cannot be complemented by pM1113-AtMinD at 0 μM IPTG. (E) HL1 mutant (ΔMinDE) complemented by pM1113-AtMinD at 50 μM IPTG. (F) HL1 mutant

(ΔMinDE) containing pM1113-MinD at 20 μM IPTG. (G) RC1 mutant (ΔMinCDE). (H) RC1 mutant (ΔMinCDE) containing pM1113-AtMinD at 50 μM IPTG. CAL-101 research buy Arrows in (B, D, G and H) mark the minicells. The bar in (A to E, G and H) represents 10 μm; the bar in (F) represents 20 μm. The sequences L-NAME HCl of the MinD in bacteria are similar to those in plants [17]. Members of the MinD family have important roles in positioning the FtsZ ring and the division apparatus to either the mid-cell of bacteria or the mid-site of chloroplasts [9]. The complementation of E. coli HL1 mutant (ΔMinDE) by AtMinD and the requirement of EcMinC for this complementation suggest that the function of MinD is also conserved between bacteria and plants. However, this complementation doesn’t require the presence of EcMinE suggests that AtMinD may have some characters different from that of EcMinD. AtMinD is localized to puncta in E. coli and chloroplasts To understand the function of AtMinD in E. coli, AtMinD-GFP and GFP-AtMinD were expressed in HL1 mutant (ΔMinDE) (Figure 2D, E, G and 2H). Similar to AtMinD, AtMinD-GFP and GFP-AtMinD can complement the minicell phenotype of HL1 mutant (ΔMinDE) with 50 μM IPTG (Table 1 and Table 2).

The KR domain reduces carbonyl groups at a specific position of the polyketide chain, and the ARO and CYC domains control chain folding by catalyzing one or more regiospecific cyclization in the polyketide chain. Typical primary products

of these type II PKSs are polyphenols that can be classified into 7 polyketide chemotypes: linear click here tetracyclines, anthracyclines, benzoisochromanequinones, tetracenomycins, aureolic acids, and angular angucyclines, as well as a group of pentagular polyphenols [4]. Additional modification by several elaborate tailoring enzymes such as find more dimerases, P450 monooxygenases, methyltransferases, and glycosyltransferases can further diversify phenolic polycyclic compounds such as actinorhodin [5]. Figure 1 Schematic diagram depicting the activity of type II PKS domains with actinorhodin biosynthesis as an example. Heterodimeric KS and CLF domains catalyze chain

AZD3965 initiation and elongation through decarboxylative condensation of malonyl building blocks, an ACP domain delivers malonyl building blocks to the KS-CLF, and a MCAT domain supplies malonyl groups to the ACP domain. The collective action of these type II PKS domains lead to the formation of highly reactive poly-β-keto intermediates. This nascent polyketide chain is modified into a specific folding pattern by tailoring enzyme domains such as those of KR, ARO, and CYC. The KR domain reduces carbonyl group at a specific position of the polyketide chain, and the ARO and CYC domains control chain folding by catalyzing one or more regiospecific cyclization in the polyketide chain. Whereafter

polyketide chain is modified by various tailoring enzymes into actinorhodin. Currently, a vast majority of polyketides is derived from a single Actinomycetes genus, Streptomyces[6]. It is difficult to culture most microorganisms on earth that produce aromatic polyketides, under standard laboratory conditions because of their different growth rates and difficulties in laboratory manipulation [7]; MRIP this evidences the fact that there are a few aromatic polyketide producers and that the complete realm of these microorganisms remains to be explored. Furthermore, studies on type II PKSs and their polyketides have been performed on a limited number of genomes. However, the current progress of computational methods and substantial increase of genome sequencing data has created new possibilities to comprehensively characterize polyketide-producing genomes and increase the number of valuable resources in this field [8]. In order to discover novel aromatic polyketides based on genome mining, it is essential to comprehensively analyze various type II PKSs in different organisms to detect type II PKSs and analyze the correlation between domain organizations and polyketide structures.

Pharm Res 2007, 24: 1720–1728.CrossRefPubMed 15. Mistry P, Stewart AJ, Dangerfield W, Okiji S, Liddle C, Bootle D, Plumb JA, Templeton D, Charlton P: In vitro and in vivo reversal of P-glycoprotein-mediated

multidrug resistance by a novel potent modulator, BAY 63-2521 solubility dmso XR9576. Cancer Res 2001, 61: 749–758.PubMed 16. Fox E, Bates SE: Tariquidar (XR9576): a P-glycoprotein drug efflux pump inhibitor. Expert Rev Anticancer Ther 2007, 7: 447–459.CrossRefPubMed 17. Lepper ER, Hicks JK, Verweij J, Zhai S, Figg WD, Sparreboom A: Determination of midazolam in human plasma by liquid chromatography with mass-spectrometric detection. J Chromatogr B Analyt Technol Biomed Life Sci 2004, 806: 305–310.CrossRefPubMed 18. Beppu K, Jaboine J, Merchant MS, Mackall CL, Thiele CJ: Effect of imatinib mesylate ARS-1620 mw on neuroblastoma tumorigenesis and vascular endothelial growth factor expression. J Natl Cancer Inst 2004, 96: 46–55.CrossRefPubMed 19. Bihorel S, Camenisch G, Lemaire M, Scherrmann JM: Modulation of the Brain Distribution of Imatinib and its Metabolites in Mice by Valspodar, Zosuquidar and Elacridar. Pharm Res 2007, 24 (9) : 1720–8.CrossRefPubMed 20. Choo EF, Kurnik D, Muszkat M, Ohkubo T, Shay SD, Higginbotham JN, Glaeser H, Kim RB, Wood AJ, Wilkinson GR: Differential in vivo sensitivity to inhibition of P-glycoprotein located in lymphocytes, PX-478 testes,

and the blood-brain barrier. J Pharmacol Exp Ther 2006, 317: 1012–1018.CrossRefPubMed 21. Dantzig AH, de Alwis DP, Burgess M: Considerations in the design and development of transport inhibitors

as adjuncts to drug therapy. Adv Drug Deliv Rev 2003, 55: 133–150.CrossRefPubMed Competing interests The authors declare that they have no competing interests. Authors’ contributions ERG participated in study design, http://www.selleck.co.jp/products/Staurosporine.html performed analytical and animal experiments, carried out all statistical analyses and drafted the manuscript. NFS participated in study design and performed animal experiments. WDF participated in study design and helped to draft the manuscript. AS participated in study design, performed animal experiments and helped to draft the manuscript. All authors approved the final manuscript.”
“Background Cervical cancer is the most common malignant gynecological cancer, and lymphatic metastasis is one of the most important metastatic routes of this cancer. The involvement of the lymphatic node is usually one of the factors predicting the prognosis. Along with the development of specific markers of lymphatic endothelium [1] and the improvement of isolation techniques for lymphatic endothelial cells [2], the role of tumor lymphangiogenesis in the metastasis of early-stage cervical carcinoma is gradually becoming a research focus. Even so, the mechanism of the tumor lymphatic metastasis is still largely unknown and there is a great deal of debate over various aspects of research in the field.

1% BSA before plating cells. Plates were again washed with PBS and air-dried. SMMC-7721 cells were preincubated with CXCL12 (100 ng/ml) for 24 h at 37°C. A cell suspension containing 2 × 105 cells/ml was prepared in serum free media. The cell suspension (150 μl) was added to the inside of each well (BSA-coated wells were provided as a negative control).

Cells were allowed to attach for 1 h at 37°C. Subsequently, unattached cells were removed by gentle washing 3 times with PBS. Then the attached cells were stained with 1% crystal violet. Each well was gently washed 3 times with selleck screening library PBS. The total crystal violet bound to the cells was eluted with 10% acetic acid and measured by the absorbance at 560 nm. All the experiments were repeated 3 times in duplicate wells. ELISA for VEGF SMMC-7721 cells were plated in 24-well tissue culture plates at a density of 1 × 105 cells per well and followed with serum starvation for 24 h with RPMI-1640. Then, cells were treated with recombinant human CXCL12 (100 ng/ml)(Peprotech, UK), and the supernatants were collected 24 h after treatment. VEGF concentration was determined using Quantikine

ELISA kits according to the manufacturer’s instructions (R&D Systems, Minneapolis, MN). In vitro tube formation coculture assay To perform the tube formation assay, Transwell chambers were precoated with growth factor-reduced BMN673 SN-38 research buy Matrigel (200 μL of 10 mg/mL). Control, NC and CXCR7 shRNA transfected cells were seeded at a density of 2 × 104 cells/well in 24-well plates and cultured for 24 h respectively. HUVECs (2 × 104 cells/well) were then seeded in Transwell chambers precoated with the Matrigel. Subsequently, Transwell chambers containing HUVECs were inserted into the 24-well plates and cocultured for 24 h. After 24 h of cocultured at 37°C and 5% CO2, the number of capillary-like tubes from three randomly chosen fields was counted and photographed under an Nikon inverted microscope (Japan). Immunohistochemistry and quantitation of microvessel density Immunohistochemistry was used to analyze

the expression of CXCR7 and CD31. Paraffin-embedded human hepatocellular carcinoma tissues were sectioned at 5 μm thickness. Tumors established in nude mice were isolated and fixed GPX6 in 4% paraformaldehyde, embedded in paraffin, and cut in 6 μm sections. Tumor sections were deparaffinized, rehydrated, and quenched with 3% hydrogen peroxide for 10 min at room temperature. The sections were incubated in protein blocking solution (5% normal horse serum, 1% goat serum in PBS) for 10 min before the addition of the primary antibody. The sections were incubated for 2 h at 37°C with rat antimouse CD31 (BD Biosciences, USA) or rabbit antihuman CXCR7 (Abcam, UK) at 1:100 dilutions. After incubation, the sections were washed in PBS for 10 min, and anti-mouse or anti-rabbit secondary biotinylated antibody was applied.

32 μmol/L) than our study (more than 2.0 μmol/L), which included middle to older aged subjects (46.1 ± 9.02 years in control, 56.7 ± 15.42 years in VC, 46.2 ± 11.35 years in exercise with VC, and 49.5 ± 15.9 year in exercise).However, the TAC levels appeared similar. For NOx with nitrite levels in all smokers; 25.23 ± 1.11 in control, 24.23 ± 2.12 μmol/L in VC, 28.23

± 1.45 μmol/L in exercise with VC, and 25.23 ± 1.30 μmol/L in exercise (Figure 3 left). Previous study in healthy, sedentary, younger (22.5 ± 3.45 years) or older individuals (65.7 ± 6.14 years) noted mean levels lower levels which were slightly lower but similar to our values (23.78 ± 5.72 μmol/L and 22.17 ± 6.14 μmol/L) [41]. The www.selleckchem.com/products/verubecestat.html higher nitrite levels in our study may be related to the high level MLN2238 nmr of PrOOH (Figure 2 right). Many reports show that NOx can react and damage protein. For example, selleck inhibitor Ischiopoulos and al-Mehdi [42] showed that peroxynitrite was generated by the reaction of NOx with superoxide and has a direct effect on tryptophan and cysteine, including protein fragmentation. Previous study in smokers showes the high level of oxidized protein compared to nonsmokers [43]. Intervention: Oxidative Stress Oxidative stress values changes with the intervention in all groups except for group 4. In Group 1, MDA, PrOOH, and NOx significantly decreased, whereas TAC increased. In Group

2, MDA and PrOOH decreased, with no other changes noted. In Group 3, MDA, PrOOH, NOx, TAC, and beta-endorphin levels all increased significantly Figure 3 shows the plasma NOx levels after the 2 month intervention, and results showed an improved NOx level in group 3 (32.34 ± 2.78 μmol/L) and a slightly increased level in group 2 (1.23 ± 2.12 μmol/L), Thalidomide but it was lower than in a previous study by Franco [41], which showed higher levels

of NOx in both healthy younger (44.73 ± 6.48 μmol/L) and older subjects (45.88 ± 9.84 μmol/L). Physiologically, a lower level of NOx can be indicative of a depressed function in nitric oxide synthase (NOS) and lower release of NOx in the smoker’s plasma, which can cause hypertension or stroke in the long term [44]. Fortunately, results in our study showed an increasing level of NOx in group 2 and 3, which might aid overall cardiovascular health. We also noted improvement of TAC (statistically) in all groups, excepted group 4 (VC > exercise and VC > exercise, alone). A previous study showed the antioxidant activity of VC flowers in arthritis-induced rats [31], which corresponded to a reduction in lipid peroxide in the liver, plasma and spleen, and also an increase in glutathione in the blood. Intervention: β-endorphin and CO Although this study was carried out in a small group of smokers, the results related to β-end showed a significant increase after strenuous exercise (Figure 5). The β-end level in this study was nearly the same as the mean value (79.46 ± 6.31 pg/ml) of a previous study [45] of smokers who consumed less than 10 cigarettes per day.

Next, we determined whether one, both, or neither of the putative RDFs uncovered by our bioinformatic analysis are required for VPI-2 excision. To do this, we constructed in-frame deletion mutations in each gene to create mutant selleck products strain SAM-3 (ΔvefA) and SAM-4 (ΔvefB). The two mutant strains and the wild-type N16961 were each inoculated into LB and all three strains grew similarly indicating that the mutant constructs did not have any general growth defect (data not shown). We determined the attB levels using QPCR in strain SAM-3

compared DMXAA to the wild-type strain grown under the same conditions. We found that no VPI-2 excision occurs in SAM-3 cells when compared with the wild type, indicating that a functional Trichostatin A copy of vefA is essential for efficient excision of VPI-2 (Figure 5). We complemented SAM-3 with a functional copy of vefA (SAM-5) and measured attB levels in these cells with the wild type levels both under standard conditions, to find that some excision occurred, but it was less than in wild-type cells (Figure 5). In our

vefB mutant strain (SAM-4), we found no difference in VPI-2 excision levels compared to wild-type grown under the same conditions, which demonstrates that vefB is not essential for excision (Figure 5). From these data it appears that vefA is the cognate RDF for VPI-2 excision. In our control experiments, transformation of SAM-3 with pBAD33 alone (resulting in strain SAM-13) did not affect attB levels (data not shown). Vibrio species island-encoded integrases with corresponding RDFs Given that our initial search for RDFs within one V. cholerae genome (strain N16961) yielded three putative RDFs (VC0497, VC1785, and VC1809), we decided to investigate further the occurrence of RDFs among Vibrio species whose genome sequence is available in the database. We performed BLAST searches against the 20 Vibrio species in the genome database, and we uncovered a total of 27 putative RDFs (Table

3). Next, we identified putative integrases within the genomes of the RDF homologues using BLAST GABA Receptor search analysis by using IntV2 as a seed. For each of the RDFs identified among the 27 genomes encompassing 10 different Vibrio species (V. cholerae, V. coralliilyticus, V. furnissii, V. harveyi, V. parahaemolyticus, V. splendidus, V. vulnificus, Vibrio sp. Ex25, RC341, and MED222), we identified a corresponding integrase with greater than 40% amino acid identities to IntV2 (VC1758) (Table 3). We examined the gene context of each RDF and integrase within each of the 20 strains to determine whether the RDF and integrase were located on the same region within a strain. From these analyses, we found that each of the 27 RDFs has a corresponding integrase within approximately 100 kb of each other (Table 3). It should be noted that from table 3, only three of the strains have been annotated completely and for many of the strains examined their ORF annotation numbering is not consecutive.

For r 1=r 2=0, the wave function in the DSN exactly reduces to that of the DN. We analyzed the probability densities in the DN and in the DSN from Figures 2 and 3, respectively, with the choice of sinusoidal signal source. The probability densities in the DN given in Figure 2b,c,d oscillate with time. Moreover, their time behaviors are more

Cyclosporin A manufacturer or less distorted. The probability density, however, does not oscillate when there are no displacement and no signal of power source (see Figure 2a). The probability densities in the DSN are distorted much more significantly than those of the DN. The time behavior of probability densities of quantum states, CP-868596 mouse both the DN and the DSN, is highly affected by external driving power source. When there is no external power source( =0), the displacement of charges, specified with a certain initial condition, gradually disappears as time goes by like a classical state. The fluctuations and uncertainty products of charges and currents are derived in the DSN, and it is shown that their value is independent of the size of the particular solutions q j p (t) and p j p (t). From this, together with the fact that q j

p (t) and p j p (t) are determined by the characteristics of , it is clear that the electric power source does not affect on the fluctuation of canonical variables. If we ignore the time dependence of Megestrol Acetate F j (t) and , decrease exponentially with time, whereas increase exponentially. From Equations 64 and 65, we can see that the time behavior of q j is determined

by two factors: One is displacement and the other is the signal of power source. For better understanding of this, recall that the amplitude of complementary functions gives displacement of the system, and the particular solutions are closely related to external driving force (i.e., in this case, the power source). In this paper, we did not consider thermal effects for the system. The thermal effects, as well as dissipation, may be worth to be considered in the studies of quantum fluctuations of Fludarabine supplier electronic circuits with nanosize elements because the practical circuits are always working in thermal states with the presence of damping. It may therefore be a good theme to investigate DSNs with thermalization as a next task, and we plan to investigate it in the near future. Appendix 1 The eighth formula of 7.

Primary outcome measures included measures of pain and functional MM-102 price capacity. Secondary outcome measures included weight loss and body composition; serum blood and hormones; and, measures of quality of life. All participants were tested for changes in energy intake; anthropometrics; body composition; resting energy expenditure; cardiovascular and muscular fitness; balance and functional capacity; serum and whole blood clinical

markers; hormonal profiles; pain selleck chemical indices; and, psychosocial parameters after 0, 10, and 14 weeks of training, dieting, and supplementation. Participants This research protocol was reviewed and approved by the university Institutional Review Board prior to initiation. Participants were recruited through area physicians, advertisements in local newspapers, campus flyers, and Internet advertisements. Interested participants were asked to contact the laboratory for an initial telephone pre-screening interview. General entrance criteria included being a female with physician diagnosed

CH5424802 research buy OA between the ages of 18-70 years with a body mass index (BMI) greater than 27 kg/m2 and no recent participation in a diet or exercise program. Individuals who met initial entrance criteria were invited to attend a familiarization session in which the details of the study were explained, human subject consent forms were signed, and personal and medical history information obtained. Subjects were not allowed to participate in this study if they: 1.) were pregnant, became pregnant, or had a desire for pregnancy; 2.) had any metabolic disorder including known electrolyte abnormalities, heart disease, arrhythmias, Etomidate diabetes, or thyroid disease; 3.) had a history of hypertension, hepatorenal, musculoskeletal, autoimmune, or neurological disease (other than knee OA); 4.) were taking thyroid, hyperlipidemic, hypoglycemic, or anti-hypertensive

medications; 5.) had taken ergogenic levels of nutritional supplements that may affect muscle mass (e.g., creatine, HMB), anabolic/catabolic hormone levels (e.g., DHEA), or weight loss supplements (e.g., thermogenics) within three months prior to the start of the study; 6.) were ingesting any anti-inflammatory products two weeks before the start of the study or additional products during the study; 7.) reported any unusual adverse events associated with this study in which the supervising physician recommended removal from the study; 8.) had significant injury or surgery to the lower extremity or spine within the last six months; 9.) did not indicate a minimal amount of perceived pain and physical function limitation on inventories used in the study; 10.) had severe arthritis that required surgery and greatly limited functionality (inability to perform lunge); or, 11.) had arthritis that required the current use of physiotherapy modalities.