A critical evaluation of salivary testosterone as a method for the assessment of serum testosterone

Although salivary testosterone (T) is often used in clinical studies accuracy is mostly questionable. State of the art data for men is sparse and for women absent. Our objective was to perform a critical evaluation of salivary T (Sal-T) as a method for indirect assessment of serum T using state of the art methods. Saliva was collected via ‘Salivette’ and ‘passive drooling’ methods. Sal-T and free T in serum after equilibrium dialysis were measured by LC-MS/MSResultsEvaluation of Sal-T results versus free T by equilibrium dialysis (ED-T) for men gave: ‘Salivette’ Sal-T = 0.05 + 0.88x ED-T, r = 0.43; ‘passive drooling’ Sal-T = 0.17 + 0.91x ED-T r = 0.71. In women, correlation was comparable but values are higher than free T: ‘passive drooling’ Sal-T = 0.12 + 2.32x ED-T, r = 0.70. The higher than expected T values in saliva, appear to be explained by T binding to salivary proteins. Iso-electric focusing of saliva proteins, followed by fractionation and LC–MS/MS assay of T showed marked testosterone peaks at pH 5.3 and 8.4, providing evidence for T binding in saliva to proteins such as albumin and proline rich protein (PRP).ConclusionsPassive drooling is the collection method of choice for testosterone in saliva. Sal-T is not directly comparable to serum free T due to T binding to saliva proteins, which substantially affects the low Sal-T in women but not the higher Sal-T in healthy adult men.     

Sex differences in jaw muscle duty factors during exercise in two environments: A pilot study

It is unknown if females and males use jaw muscles similarly during exercise. This pilot study assessed jaw elevator muscle duty factors (DFs = time of muscle activity/total recording time) at repeated sessions to test if DFs are reliable and different between sexes during exercises in two environments. Ten female and seven male subjects recruited from university soccer teams provided informed consent. Surface electromyography was recorded from masseter and temporalis muscles during biting and leg-extension laboratory exercises. Average activities to produce 20 N bite-forces for each muscle and subject determined thresholds (5–80%·T20 N) for subject-specific DF calculations during exercises performed in laboratory and natural environments. Subjects self-recorded via portable electromyography equipment during in-field leg-extension and weight-lifting exercises. Effects of variables on DFs were assessed via ANOVA (α = 0.05) and simple effects testing (Bonferroni-adjusted p ⩽ 0.012). All subjects used jaw muscles during exercises in both environments. DFs between laboratory sessions were reliable (R = 0.84). During laboratory exercises, male temporalis DFs were significantly higher than female DFs from both muscles (p ⩽ 0.001). During in-field exercises females had higher DFs during weight-lifting while males had higher DFs during leg-extensions. In-field sex differences were significant at most thresholds and showed larger effect sizes for leg-extension compared to weight-lifting exercises.     

Salivary vasopressin increases following intranasal oxytocin administration

Extant research has documented the effects of intranasal administration of oxytocin (OT), and to a lesser degree Arginine Vasopressin (AVP) – two structurally-related neuropeptides – on brain and behaviour, yet the effects of exogenous manipulation of one on circulating levels of the other remain unknown. Studies have shown that OT administration impacts the peripheral levels of numerous hormones; however, whether OT administration also increases AVP concentrations has not been explored. Utilizing a double-blind placebo-controlled within-subject design, ten male and female subjects provided ten saliva samples over four consecutive hours: at baseline and nine times following OT administration. Results indicate that salivary AVP increased in the first hour following OT manipulation (OT condition: mean AVP = 2.17 pg/ml, SE = 28, placebo condition: mean AVP = 1.42 pg/ml, SE = .18) but returned to baseline levels at the next assessment (80 min) and remained low for the remaining period. Similar to OT, AVP showed high degree of individual stability and baseline levels of AVP correlated with AVP concentrations at the first and second post-administration hours regardless of drug condition (Pearson r = .85–.93). Validity of salivary AVP ELISA measurement was verified by demonstrating individual stability of salivary AVP over a six-month period (r = .70, p < .000) as well correlation with plasma levels over the same period (r = .32, p = .037) in a sample of 45 young adults who did not participate in the current study. Overall, findings suggest a potential crosstalk between OT and AVP and indicate that baseline levels of the two neuropeptides may shape the degree to which these systems respond to exogenous manipulation.     

Invalidation of a commercially available human 5α-dihydrotestosterone immunoassay

Enzyme immunoassays (EIA) are commonly utilized for the evaluation of androgens in biological fluids; however, careful consideration must be given to cross-reactivity with other endogenous sex-steroid hormones. Our purpose was to determine the validity of a commonly-utilized commercially-available dihydrotestosterone (DHT) EIA. Serum samples obtained from older hypogonadal men who participated in a 12-month randomized controlled trial evaluating the effects of testosterone-enanthate (125 mg/week) or vehicle in combination with finasteride (5 mg/day) or placebo were assayed for DHT via EIA and using a validated gold-standard LC–MS/MS approach. Additionally, commercially-available (DHT-free) buffer containing graded testosterone doses was evaluated by DHT immunoassay. DHT concentrations measured via EIA were 79% to >1000% higher than values obtained by LC–MS/MS (p < 0.05), with the largest differences (415–1128%) occuring in groups receiving finasteride. Both LC–MS/MS and EIA indicated that testosterone-enanthate increased serum DHT to a similar magnitude. In contrast, finasteride-induced reductions in DHT were detected by LC–MS/MS, but not EIA (p < 0.05). No significant associations were present for DHT concentrations between measurement techniques. Cross-reactivity of testosterone with the immunoassay ranged from 18% to 99% and DHT concentrations measured by EIA were highly associated with the spiked testosterone concentrations in DHT-free buffer (r = 0.885, p < 0.001). In conclusion, we provide evidence invalidating a commonly-utilized commercially-available DHT immunoassay because significant cross-reactivity exists between testosterone and the EIA and because the changes in DHT observed via EIA were not associated with a validated gold-standard measurement technique. The cross-reactivity of testosterone is particularly concerning because testsoterone is present in 100-fold greater concentrations than is DHT within the circulation.     

A microsphere-based duplex competitive immunoassay for the simultaneous measurements of aldosterone and testosterone in small sample volumes: Validation in human and mouse plasma

BackgroundThe small blood volumes available in rodent studies often limit adequate quantification of all hormones of interest. We report here the development of two new assays combining an extraction step with multiplex immunoassay (MIA) technology for the simultaneous determination of aldosterone and testosterone in 50 μl sample volume.MethodsFollowing solvent extraction, aldosterone and testosterone competitive immunoassays are performed incorporating biotinylated tracers and antibody-coated beads each having a unique fluorescence. Quantification is via addition of streptavidin–R–phycoerythrin (SA–PE). The assays were validated and compared to established methods. Baseline hormone levels in mice from four different strains, and changes after ACTH and HCG stimulation in CD-1 mice are shown.ResultsThe assays are sensitive (aldosterone 15 pg/ml, testosterone 12 pg/ml), reproducible (intra-/inter-assay imprecision aldosterone 5.1–15.6%/9.9–15.8% and testosterone 9.7–10.9%/7.7–11.4%) and correlate significantly to established assays (r = 0.94–0.95). Baseline aldosterone levels varied between strains, but not between the genders. Testosterone was significantly higher in male of all strains except in C57BL/6× NMRI mice. After ACTH injection, aldosterone (median, interquartile range) rose from 354 (261–396) pg/ml to 2008 (875–2467) in male and from 260 (210–576) to 1120 (734–1528) in female CD-1 mice. HCG injection in the same strain increased testosterone in male mice only (3.5 (0.4–8.3) ng/ml to 31.8 (30.4–33.9) ng/ml, P < 0.01).ConclusionsWe describe a MIA for the simultaneous measurement of aldosterone and testosterone in small volumes after extraction. In addition to presenting a new tool for steroid research in rodent models, our data show strain-dependent differences in steroid hormone metabolism in rodents.     

Anthropometry and somatotypes of competitive DanceSport participants: A comparison of three different styles

Anthropometry in dance and aesthetic sports has been shown to play an important role in selection and performance criteria. The aim of the present study was to examine variations in somatotype and anthropometry in three different competitive dance styles: DanceSport Standard, Latin American and Ten Dance. Anthropometry and somatotype data were collected from thirty couples competing in Standard (n = 24 individuals), Latin American (n = 14) and Ten Dance (n = 22) styles. A single tester (ISAK Level 1) carried out all anthropometric measurements using the Heath-Carter protocol and somatotypes were calculated using the Heath-Carter's decimal equations. Results indicated that the mean somatotype for the male dancers was 2.4–3.9–3.2, whilst for females it was 2.7–2.7–3.5. Factorial analysis reported Standard dancers scored significantly higher for ectomorphy, sitting height and arm span than Latin dancers (p < 0.05). Correlation analysis with the Standard dancer's International Ranking highlighted moderate positive correlation with mesomorphy (r = 0.434, p < 0.05) and negative correlation with ectomorphy (r = −0.546, p < 0.001). The findings of this study show that somatotypes differ among DanceSport participants by dance style. Compared with other aesthetic sports, male and female dancers were less mesomorphic and more ectomorphic. Standard dancers tend to be more ectomorphic with greater height, longer arm span and greater sitting height compared with Latin American dancers. Although Standard dancers were ectomorphic, those dancers who had higher mesomorphic ranking had higher places in the dancers’ international ranking.     

Homocysteine and inflammation as main determinants of oxidative stress in the elderly

Oxidative stress is commonly observed in the elderly and could be involved in age-related diseases. However, the determinants of superoxide anion overproduction are not clearly understood. Superoxide anion production was evaluated using a lucigenin-based chemiluminescence method in 478 elderly subjects (304 women, 174 men; 79.5 ± 7.1 years). Homocysteine (HCy) metabolism (homocysteinemia, vitamin B12, plasma, and erythrocyte folates), inflammation (CRP, fibrinogen, α-1 acid glycoprotein), lipid parameters (total cholesterol, triglycerides, HDL and LDL cholesterol), and nutritional parameters (albumin, transthyretin) were determined. The results show that HCy levels (p < 0.001) and superoxide anion production (p = 0.04) increase with aging, but CRP does not. Highest HCy (> 20 μM) (OR 1.83 (1.09–3.07), p = 0.02) and CRP over 5 mg/L (adjusted OR 2.01 (1.15–3.51), p = 0.01) are the main determinants in superoxide anion production in the elderly. These clinical data are confirmed in an in vitro study using THP-1 monocyte-like cells. Incubation with HCy thiolactone (HTL) (0–200 μM) and LPS (0–20 ng/ml) dramatically enhances NADPH oxidase expression and activation. Moreover, a synergic action was evidenced for low concentrations of HTL (20 μM) and LPS (5 ng). Taken together, the clinical data and in vitro experiments support the hypothesis that moderate homocysteinemia and low-grade inflammation synergically enhance NADPH oxidase activity in the elderly.     

The interface of hypothalamic–pituitary–adrenocortical axis and circulating brain natriuretic peptide in prediction of cardiopulmonary performance during physical stress

Brain natriuretic peptide (NT-pro-BNP) was implicated in the regulation of hypothalamic–pituitary–adrenocortical (HPA) responses to psychological stressors. However, HPA axis activation in different physical stress models and its interface with NT-pro-BNP in the prediction of cardiopulmonary performance is unclear. Cardiopulmonary test on a treadmill was used to assess cardiopulmonary parameters in 16 elite male wrestlers (W), 21 water polo player (WP) and 20 sedentary age-matched subjects (C). Plasma levels of NT-pro-BNP, cortisol and adrenocorticotropic hormone (ACTH) were measured using immunoassay sandwich technique, radioimmunoassay and radioimmunometric techniques, respectively, 10 min before test (1), at beginning (2), at maximal effort (3), at 3rd min of recovery (4). In all groups, NT-pro-BNP decreased between 1 and 2; increased from 2 to 3; and remained unchanged until 4. ACTH increased from 1 to 4, whereas cortisol increased from 1 to 3 and stayed elevated at 4. In all groups together, ΔNT-pro-BNP2/1 predicted peak oxygen consumption (B = 37.40, r = 0.38, p = 0.007); cortisol at 3 predicted heart rate increase between 2 and 3 (r = −0.38,B = −0.06, p = 0.005); cortisol at 2 predicted peak carbon-dioxide output (B = 2.27, r = 0.35, p < 0.001); ΔACTH3/2 predicted peak ventilatory equivalent for carbon-dioxide (B = 0.03, r = 0.33, p = 0.003). The relation of cortisol at 1 with NT-pro-BNP at 1 and 3 was demonstrated using logistic function in all the participants together (for 1/cortisol at 1 B = 63.40, 58.52; r = 0.41, 0.34; p = 0.003, 0.013, respectively). ΔNT-pro-BNP2/1 linearly correlated with ΔACTH4/3 in WP and W (r = −0.45, −0.48; p = 0.04, 0.04, respectively). These results demonstrate for the first time that HPA axis and NT-pro-BNP interface in physical stress probably contribute to integrative regulation of cardiopulmonary performance.     

An effective technique for the processing of saliva for the analysis of leptin and adiponectin

The recovery of protein from saliva has been extensively investigated as a method to monitor health. The aim of this study was to compare filtration and centrifugation as two methods of saliva processing necessary for determining the levels of salivary leptin and adiponectin. Thirty-seven healthy patients (median age of 45 years; range 35–73) participated in the study. Unstimulated whole saliva was collected by a drooling technique. An aliquot was filtered using a Millex-Millipore^® (0.45 μm PVDF Dura Pore membrane) syringe and a second aliquot was centrifuged at 15 000 × g for 15 min at 4 °C. Leptin and adiponectin levels were analyzed using an ELISA kit for serum (RayBio^®, GA, USA) with minor modifications. Leptin and adiponectin levels following the filtration technique yielded comparable results with those after centrifugation. Correlation was observed between filtered and centrifuged salivary leptin levels ((r = 0.9155; 95% CI 0.8362–0.9573; p < 0.0001) with concordance correlation coefficient k 0.9114 (95% CI 0.8332–0.9539)). Less correlation was observed for adiponectin ((r = 0.5718; 95% CI 0.3041–0.7558; p = 0.0002) with concordance correlation coefficient k 0.5586 (95% CI 0.2977–0.7419)). Using a Bland–Altman plot, similar measurements for both adipocytokines were observed with mean difference within a 95% CI, and interpreted as no systematic differences between the two processing techniques. This study showed that filtration is an alternative saliva processing technique to retrieve supernatant for protein analysis. Filtered saliva yielded leptin and adiponectin concentrations comparable with those obtained from centrifuged saliva.     

The influence of a TNF gene polymorphism on the severity of rheumatoid arthritis in the Brazilian Amazon

PurposeThe aim of this study was to investigate the influence of the TNF -308 G/A polymorphism in the promoter region of the tumor necrosis factor-α gene on the susceptibility and severity of rheumatoid arthritis (RA) in individuals from the Brazilian Amazon.MethodsA total of 323 individuals—192 healthy controls without arthritis and 131 individuals suffering from arthritis—were genotyped for this polymorphism using a methodology based on PCR-RFLP.ResultsThe frequency of the A allele (TNF2) in rheumatoid arthritis sufferers was not significantly higher than in the controls (p = 0.926; OR = 0.97; confidence interval 0.54–1.76). However, using a logistic regression model, when the patients were stratified according to whether the manifestations were preponderantly articular or systemic, there was a strong association between the TNF2 allele and systemic arthritis (p = 0.001; OR = 5.89; confidence interval = 1.98–17.5) as well as the use of anti-TNF immunotherapy (p = 0.023; OR = 1.10; confidence interval = 1.00–1.14). The main factors that were found to influence the risk of extra-articular disease were age greater than or equal to 60 years (p = 0.008; OR = 4.06; confidence interval = 1.45–11.38), disease duration greater than 10 years (p = 0.031; OR = 3.10; confidence interval = 1.11–8.63) and positive rheumatoid factor (p = 0.035; OR = 2.07; confidence interval = 1.05–4.09).ConclusionsThese results suggest that the TNF2 allele is associated with the more serious forms of the disease in individuals from the Brazilian Amazon but not with a risk for developing RA.     

Differing patterns of peripheral blood leukocyte telomere length in rheumatologic diseases

Telomeres progressively shorten with repeated somatic tissue cell division, their length being an indicator of cellular ageing. Telomeric dysfunction may be implicated in a variety of diseases. We measured mean telomere length in peripheral blood leukocytes (PBL) from patients with various rheumatologic diseases. Mean PBL telomere length was measured using real-time quantitative polymerase chain reaction (Q-PCR) assay in a control population (n = 130; age range: 3–94 years) and in subjects diagnosed with rheumatoid arthritis (RA; n = 86; age range: 31–82 years), psoriatic arthritis (PA; n = 56; age range: 26–79 years) and ankylosing spondylitis (AS; n = 59; age range: 21–75 years). These diseases are associated with chronic systemic inflammatory activity. Telomere length was also quantified in subjects with osteoarthritis (OA; n = 34; age range: 43–82 years) and osteoporosis (OP; n = 35; age range: 59–95 years), diseases without a chronic systemic inflammatory component. Telomere length in OA showed no differences from age-matched controls (p = 0.234), but was significantly shorter in OP (p = 0.001). Telomere length was significantly longer than controls in RA (p = 0.015), PA (p < 0.001) and AS (p < 0.001). Different patterns in telomere length from PBL are evidenced in rheumatologic pathologies, possibly dependent on the presence or absence of chronic systemic inflammation.     

A study on the association of TNF-α-308, IL-6-174, IL-10-1082 and IL-1RaVNTR gene polymorphisms with rheumatic heart disease in Pakistani patients

Inflammation is an important contributor to the pathogenesis of rheumatic heart disease (RHD), a disorder of heart valves caused by a combination of immune, genetic and environmental factors. Cytokines are important mediators of inflammatory and immune responses. The aim of this study was to investigate the role of cytokine gene polymorphisms and their potential usefulness as biomarkers in RHD patients from Pakistan. We screened 150 RHD patients and 204 ethnically matched controls for tumor necrosis factor (TNF)-α^-308G/A, interleukin (IL)-10^?1082 G/A, interleukin (IL)-6^-174 G/C and a variable number of tandem repeats (VNTRs) polymorphism of the IL-1Ra gene using polymerase chain reaction. The results showed that TNF-α^-308 A and IL-6^-174 G alleles were associated with susceptibility to RHD (p = 0.000; OR = 2.81; CI = 1.5–5.14 and p = 0.025; OR = 1.50; CI = 1.04–2.16 respectively). The TNF-α^-308 AA and GA genotypes were associated with susceptibility to RHD (p = 0.012; OR = 9.94; CI; 1.21–217.3 and p = 0.046; OR = 1.97; CI = 0.98–3.97 respectively) while the GG genotype seemed to confer resistance (p = 0.003; OR = 0.39; CI = 0.20–0.76). The GG genotype for IL-6^-174 was significantly associated with predisposition to RHD (p = 0.015; OR = 2.6; CI = 1.17–5.85). The A1 (four repeats) and A2 (two repeats) alleles at the IL-1Ra VNTR polymorphism were associated with resistance and susceptibility to RHD respectively. However, this polymorphism deviated from Hardy–Weinberg equilibrium in both patients and controls in our population. TNF-α^-308 and IL-6^-174 polymorphisms may be useful markers for the identification of individuals susceptible to RHD in Pakistan. These individuals could be provided aggressive prophylactic intervention to prevent the morbidity and mortality associated with RHD.     

Chromatin structure analysis of spermatozoa from reciprocal chromosome translocation (RCT) carriers with known meiotic segregation patterns

The presence of reciprocal chromosome translocations (RCTs), as well as sperm chromatin disturbances, is known to exert negative influence on male fertility. The aim of this study was to identify an association between chromosome structural rearrangements in male RCT carriers and sperm seminological parameters (concentration, motility, morphology), chromatin status (fragmentation and maturity), meiotic segregation pattern and observed chromosomal hyperhaploidy. Sperm samples originated from ten male RCT carriers with reproductive failure/success. TUNEL assay (DNA fragmentation) and chromomycin A3 (CMA3)/aniline blue (AB) staining (chromatin maturity) were used to analyze sperm chromatin status while fluorescent in situ hybridization (FISH) was applied to observe meiotic segregation patterns and hyperhaploidy in spermatozoa. We found that the mean level of sperm DNA fragmentation in the RCT carrier group (18.0 ± 11.9%) was significantly higher (p = 0.0006) than the mean of the control group (7.5 ± 4.3%). There was no correlation observed between sperm DNA fragmentation levels (5.6–38.0%) and the frequency of genetically normal/balanced gametes (34.3–62.4%), sperm seminological quality or revealed reproductive failure. In contrast, a correlation between the frequencies of genetically normal/balanced spermatozoa and of gametes with mature chromatin was observed (CMA3: R = 0.4524, p = 0.2604; AB: R = 0.5238, p = 0.1827). A statistically significant increase in the hyperhaploidy level of selected chromosomes in all analyzed RCT carriers was documented but was not correlated to sperm seminology or fertility status. Further evaluation and additional assays toward sperm chromatin quality assessment in RCT carriers is suggested to explain the complexity of genomic structural rearrangements and its possible relevance to reproductive success or failure.     

Relationship between zinc levels and plasma leptin in hemodialysis patients

Recent evidences suggested a possible relationship between zinc deficiency and leptin levels in pathogenesis of anorexia in chronic kidney disease. The present study addressed the relationship between zinc and leptin in hemodialysis (HD) patients.MethodsFifty HD patients (54.3 ± 12.7 years old, 62% men) were studied and compared to 21 healthy volunteers (50.7 ± 15.7 years old, 43% men). Biochemical data, serum zinc, plasma leptin, IL-6, TNF-α and C-Reactive Protein levels were determined. Anthropometric parameters, food intake and appetite score were also assessed.ResultsThe leptin levels were higher in HD patients (16.1 μg/mL (0.21–118.25) vs 6.0 μg/mL (0.50–23.10)) in healthy volunteers (p = 0.04), whereas serum zinc levels were lower (54.5 ± 16.3 μg/dL) compared to healthy volunteers (78.4 ± 9.4 μg/dL) (p = 0.0001). The plasma leptin was correlated negatively with plasma zinc (r = ?0.33; p = 0.007), energy (r = ?0.38; p = 0.002) and protein intake (r = ?0.34; p = 0.006) and, positively correlated with BMI (r = 0.54; p = 0.0001), % body fat (r = 0.70; p = 0.0001) and conicity index (r = 0.46; p = 0.001). Plasma zinc was associated with hemoglobin (r = 0.30; p = 0.04) and negatively associated with TNF-α (r = ?0.37; p = 0.002) and C-Reactive Protein (r = ?0.37; p = 0.004). There was no correlation among Zn, leptin and appetite score in these patients.ConclusionThis study showed that low plasma zinc levels are negatively associated with high leptin levels in HD patients.     

The relationship between fetal and maternal selenium concentrations in sheep and goats

In this study, biological samples (slaughterhouse material) were collected from 30 sheep and 36 goats and classified according to gestational stage into either early or late gestation. Samples consisted of allantoic fluid, amniotic fluid, fetal liver, fetal kidney, fetal thyroid gland, maternal plasma and liver to determine selenium (Se) concentrations throughout gestation. The Se concentrations in the allantoic fluid, fetal liver and kidney increased significantly (p < 0.01) during late gestation. Concurrently, the Se concentrations in amniotic fluid, maternal plasma and liver decreased significantly (p < 0.01) over time. Significant (p < 0.01) positive relationships were recorded between the age of the fetus and Se concentrations in the allantoic fluid (r = 0.57–0.75), fetal liver (r = 0.43–0.59) and kidney (r = 0.80–0.81) in both sheep and goats. A significant (p < 0.05) positive relationships were also recorded between the Se concentrations in the allantoic fluid and fetal liver (r = 0.35–0.37), the maternal plasma and liver Se concentrations (r = 0.37–0.57) between sheep and goats. A significant (p < 0.05) negative correlation was recorded between the Se concentrations in the allantoic fluid with maternal plasma of sheep (r = −0.41) as well as between the fetal liver and maternal liver Se (r = −0.22 to 0.50) and a negative correlation (r = −0.42 to 0.43) (p < 0.01) between Se concentrations in the fetal liver and amniotic fluid in both sheep and goats, respectively. Se concentration in the fetal liver was significantly (p < 0.01) higher than that of the kidney and thyroid. In the thyroid gland no morphological differences were noted. Strong fetal–maternal relationships in Se concentration were evident throughout the gestational period and dams seem to sacrifice Se levels in order to maintain that in the fetus. Se concentrations in the amniotic and allantoic fluids could be used as a possible indicator of the Se status of the fetus throughout gestation.     

Effects of short- and long-term changes in spatial density on the social behavior of captive chimpanzees (Pan troglodytes)

The constraints of captivity may often require non-human primates to experience restrictions in space for both long- and short-term periods of time. The tension-reduction model predicts that great apes should increase affiliative behaviors and decrease aggressive behaviors as a coping strategy. The conflict avoidance model, however, predicts that great apes should decrease all social interactions (affiliative and aggressive) as a coping strategy. The purpose of this study was to test the conflict-avoidance and tension-reduction models by examining the effects of both short-term (1–2 days) and long-term (6 months) changes in spatial density on social behavior in 23 adult captive chimpanzees (6 males, 17 females) housed at the Primate Foundation of Arizona. Affiliative (i.e., social groom, social play) and agonistic (i.e., charging display, attack) were assessed using scan-sampling techniques while subjects were rotated from high-density to low-density conditions for varying lengths of time. Results of short-term increases in spatial density (crowding) supported the conflict-avoidance strategy for females, through reduced levels of aggressive (F_1,16 = 17.11, p = 0.001) and affiliative (F_1,16 = 21.13, p < 0.001) behaviors. Males, however, supported the tension-reduction model during short-term high-density by decreasing aggression (F_1,5 = 10.53, p = 0.02) while increasing affiliation (F_1,5 = 9.10, p = 0.03). Females partially supported the tension-reduction model during long-term high-density by increasing affiliative behaviors (F_1,16 = 14.19, p = 0.002) compared to short-term high-density levels, while rates of aggression remained low. Finally, males supported the tension-reduction by reducing aggression (F_1,5 = 7.668, p = 0.04) and increasing affiliation (F_1,5 = 6.08, p = 0.05) during long-term high-density. Results confirm that chimpanzees use different strategies during short-term versus long-term increases in spatial density and that these strategies may be influenced by individual sex. However, sample sizes are small and additional research on male chimpanzees is needed.     

Extracellular matrix metalloproteinase inducer expression has an impact on survival in human bladder cancer

Aim: Extracellular matrix metalloproteinase inducer (EMMPRIN) has been shown to promote tumor invasion and metastasis via stimulating matrix metalloproteinase synthesis in neighboring fibroblasts, to enhance angiogenesis via vascular endothelial growth factor, to induce chemoresistant tumor cells via the production of hyaluronan, and to confer resistance of cancer cells to anoikis through inhibition of Bim. The purpose of this study was to investigate the expression of EMMPRIN in human primary bladder cancer and to evaluate its prognostic value. Methods: EMMPRIN expression patterns were detected by immunohistochemistry. In order to determine its prognostic value, overall survival (OS) and progression-free survival (PFS) were evaluated using the Kaplan–Meier method, and multivariate analysis was performed using the Cox proportional hazard analysis. Results: Of the 101 cases with bladder cancers, 68 (67.3%) cases were positive for EMMPRIN expression. When categorized into negative vs. positive expression, EMMPRIN was associated with the stage (p = 0.006), the grade (p = 0.002), carcinoma in situ (p = 0.01), the recurrence (p = 0.009), the progression (p = 0.009), and the death (p = 0.01) of patients with bladder cancer. Moreover, positive EMMPRIN expression clearly predicted poorer PFS (p = 0.008) and OS (p = 0.006). In the multivariate analysis, positive EMMPRIN expression was an independent prognostic factor for PFS (p = 0.03) and OS (p = 0.03). Conclusion: EMMPRIN expression was greater in bladder cancers than in the adjacent normal tissues and may be a useful prognostic marker for patients with bladder cancer.     

Neutrophil-to-lymphocyte ratio is not associated with risk of sarcopenia in elderly COVID-19 patients

BackgroundTo assess the existence of association between neutrophil to lymphocyte ratio (NLR) and the risk of sarcopenia in COVID-19 patients.MethodsA retrospective cross-sectional study was conducted in a university hospital with patients with an active COVID-19 infection admitted to the nursing ward or intensive care unit (ICU) between September to December 2020. Sarcopenia risk was assessed using the Strength, Assistance for walking, Rise from a chair, Climb stairs and Falls (SARC-F). Biochemical analyses were assessed by circulating of C-reactive protein, D-dimer, neutrophils, lymphocytes count and NLR. Sixty-eight patients were evaluated and divided into tertiles of NLR values and the association between NLR and sarcopenia risk were tested using the linear regression analyses and p < 0.05 were considered as significant.ResultsSixty-eight patients were evaluated and divided in NLR tertiles being the 1st (men = 52.2%; 71.1 ± 9.0 y; NLR: 1.1–3.85), 2nd (women = 78.3%; 73.2 ± 9.1 y; NLR: 3.9–6.0) and 3rd (men = 72.7%; 71.7 ± 10.4 y; NLR: 6.5–20.0). There was a difference between the tertiles in relation to the first to the biochemical parameters of total neutrophils count (p = 0.001), C-reactive protein (p = 0.012), and D-dimer (p = 0.012). However, no difference was found in linear regression analysis between tertiles of NLR and SARC-F, if in total sample (p = 0.054) or divided by sex, if men (p = 0.369) or women (p = 0.064).ConclusionIn elderly patients hospitalized with COVID-19, we do not find an association between the risk of sarcopenia and NLR.     

Size and density of East Greenland polar bear (Ursus maritimus) skulls: Valuable bio-indicators of environmental changes?

There is an increasing interest in assessing the relationship between climatic oscillations, environmental contaminants and the modelling of animal physiological and morphological responses. We therefore undertook a study of skull condylobasal length (CBL; reflecting body size) and bone mineral density (BMD) in 87 East Greenland male polar bears (Ursus maritimus) sampled in the time period of 1892–2010. The purpose of the study was to investigate if these measures are potential candidates as indicators for stress associated with climate change and long-range transported toxic industrial chemicals. The analyses showed that both BMD and CBL in polar bears sampled in period 4 (1999–2010, n = 57) were significantly lower when compared with period 2 (1920–1936, n = 19) (both p < 0.02). Groups of persistent organohalogen contaminants (PCBs [polychlorinated biphenyls], DDT [dichlordiphenyltrichlorethane], HCH [hexachlorocyclohexane], HCB [hexachlorobenzene], chlordanes, dieldrin, PBDEs [polybrominated diphenyl ethers]) were measured in period 4 and multiple regression analyses controlling for age showed that dieldrin had a significant negative effect on BMD (p = 0.03, n = 52) while significant positive correlations with CBL were found for DDT, dieldrin and PBDE (all p < 0.05, n = 52). When testing the correlation with the North Atlantic Oscillation climate index no significant relationship was found for BMD (p = 0.97, r = −0.01, n = 27) nor CBL (p = 0.31, r = −0.2, n = 27). We therefore suggest that BMD and body size have decreased in East Greenland polar bear males over the past 120 years and that exposure to organohalogen contaminants may explain the BMD reductions. It is, however, not entirely clear if and how climatic oscillations affected the reductions in body size and BMD mainly because of the limited sample size in period 2 and lower mean age in period 4. Therefore, precautions should be taken towards a final conclusion on BMD and CBL as bioindicators for climate oscillations and exposure to toxic environmental chemicals. It is recommended that the sampling and archiving of East Greenland polar bear skulls continue in order to further explore how CBL and BMD reflect individual and population response upon exposure to environmental stress.