Preferential production of G-CSF by a protein-like Lactobacillus rhamnosus GR-1 secretory factor through activating TLR2-dependent signaling events without activation of JNKs.

BACKGROUND: Different species and strains of probiotic bacteria confer distinct immunological responses on immune cells. Lactobacillus rhamnosus GR-1 (GR-1) is a probiotic bacterial strain found in both the intestinal and urogenital tracts, and has immunomodulatory effects on several cell types including macrophages. However, detailed immunological responses and the signaling mechanism involved in the response are largely unknown. RESULTS: We examined the production of GR-1-induced cytokines/chemokines and signaling events in macrophages. Among 84 cytokines and chemokines examined, GR-1 discretely induced granulocyte colony-stimulating factor (G-CSF) mRNA at highest levels (>60-fold) without inducing other cytokines such as IL-1α, IL-1ß, IL-6 and TNF-α (<5-fold). The toll-like receptor (TLR) 2/6-agonist PAM2CSK4, TLR2/1-agonist PAM3CSK4 and TLR4-agonist lipopolysaccharide induced all of these inflammatory cytokines at high levels (>50-fold). The TLR2 ligand lipoteichoic acid activated all mitogen-activated kinases, Akt and NF-κB; whereas, GR-1 selectively activated extracellular regulated kinases and p38, NF-κB and Akt, but not c-Jun N-terminal kinases (JNKs) in a TLR2-dependent manner. Using specific inhibitors, we demonstrated that lack of JNKs activation by GR-1 caused inefficient production of pro-inflammatory cytokines but not G-CSF production. A secreted heat-labile protein-like molecule, 30-100 kDa in size, induced the preferential production of G-CSF. CONCLUSION: This study elucidated unique signaling events triggered by GR-1, resulting in selective production of the immunomodulatory cytokine G-CSF in macrophages.

Spontaneous abortion and preterm labor and delivery in nonhuman primates: evidence from a captive colony of chimpanzees (Pan troglodytes).

BACKGROUND: Preterm birth is a leading cause of perinatal mortality, yet the evolutionary history of this obstetrical syndrome is largely unknown in nonhuman primate species. METHODOLOGY/PRINCIPAL FINDINGS: We examined the length of gestation during pregnancies that occurred in a captive chimpanzee colony by inspecting veterinary and behavioral records spanning a total of thirty years. Upon examination of these records we were able to confidently estimate gestation length for 93 of the 97 (96%) pregnancies recorded at the colony. In total, 78 singleton gestations resulted in live birth, and from these pregnancies we estimated the mean gestation length of normal chimpanzee pregnancies to be 228 days, a finding consistent with other published reports. We also calculated that the range of gestation in normal chimpanzee pregnancies is approximately forty days. Of the remaining fifteen pregnancies, only one of the offspring survived, suggesting viability for chimpanzees requires a gestation of approximately 200 days. These fifteen pregnancies constitute spontaneous abortions and preterm deliveries, for which the upper gestational age limit was defined as 2 SD from the mean length of gestation (208 days). CONCLUSIONS/SIGNIFICANCE: The present study documents that preterm birth occurred within our study population of captive chimpanzees. As in humans, pregnancy loss is not uncommon in chimpanzees, In addition, our findings indicate that both humans and chimpanzees show a similar range of normal variation in gestation length, suggesting this was the case at the time of their last common ancestor (LCA). Nevertheless, our data suggest that whereas chimpanzees' normal gestation length is ∼20-30 days after reaching viability, humans' normal gestation length is approximately 50 days beyond the estimated date of viability without medical intervention. Future research using a comparative evolutionary framework should help to clarify the extent to which mechanisms at work in normal and preterm parturition are shared in these species.

Factors affecting wounding aggression in a colony of captive chimpanzees (Pan troglodytes).

Previous research has suggested that chimpanzees (Pan troglodytes) display higher levels of aggression in captivity than in the wild. One of the challenges of captive management, therefore, is to balance the chimpanzees' need for social interaction with managements' desire to minimize wounding and aggression. Various captive studies have examined the effects of individual and social variables on the frequency of wounding aggression, but none have examined these variables simultaneously. We collected retrospective wounding data for severe wounds from 83 captive chimpanzees (36 males, 47 females) from January 1993 to December 2003. The context of the wounding event, including individual age and sex, group age and sex composition, group duration, and portion of the week (weekday vs. weekend) were collected. Logistic regression analysis was performed to determine which variables had a significant effect on the probability of a severe wounding event. The sex and age composition of the group, group duration, and portion of the week had a statistically significant association with wounding. All-male groups (Odds Ratio (OR)=6.738) had the highest risk of wounding aggression, with uni-male groups (OR=3.311) having the next largest. Compared to individuals in all sub-adult groups, individuals in either all-adult (OR=4.516) or mixed-age (OR=3.587) groups had a higher risk of wounding. There was an inverse association between group duration and wounding (OR=0.821). Finally, there was an increased risk of wounding during the work week (OR=1.653). These results suggest that captive management should pay close attention to group composition, as well as levels of human activity, when devising strategies to reduce captive chimpanzee aggression.

Heart disease is common in humans and chimpanzees, but is caused by different pathological processes.

Heart disease is common in both humans and chimpanzees, manifesting typically as sudden cardiac arrest or progressive heart failure. Surprisingly, although chimpanzees are our closest evolutionary relatives, the major cause of heart disease is different in the two species. Histopathology data of affected chimpanzee hearts from two primate centers, and analysis of literature indicate that sudden death in chimpanzees (and in gorillas and orangutans) is commonly associated with diffuse interstitial myocardial fibrosis of unknown cause. In contrast, most human heart disease results from coronary artery atherosclerosis, which occludes myocardial blood supply, causing ischemic damage. The typical myocardial infarction of humans due to coronary artery thrombosis is rare in these apes, despite their human-like coronary-risk-prone blood lipid profiles. Instead, chimpanzee 'heart attacks' are likely due to arrythmias triggered by myocardial fibrosis. Why do humans not often suffer from the fibrotic heart disease so common in our closest evolutionary cousins? Conversely, why do chimpanzees not have the kind of heart disease so common in humans? The answers could be of value to medical care, as well as to understanding human evolution. A preliminary attempt is made to explore possibilities at the histological level, with a focus on glycosylation changes.

Reproductive aging in female chimpanzees (Pan troglodytes).

Published age-specific fertility rates document a sharp decline in female chimpanzee fertility after age 35 years. However, in contrast to data on human females, little else is known regarding reproductive aging in chimpanzees. We documented age-related changes in estrous cycling, hormone profiles, and reproductive physiology in 14 female chimpanzees (Pan troglodytes) ranging in age from 32 to 50 years. Estrous data were analyzed from daily cycle charts, averaging 14.1 years of cycle data per subject, after omission of gestational periods and postpartum amenorrhea. Concentrations of total luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol, and progesterone were assayed in serum samples taken biannually. Sample collection times were chosen to avoid the ovulatory LH and FSH peaks of the female's cycle and yielded a mean of 16.2 serum samples over an average of 14.0 years per subject. Analysis of cycle charts revealed a negative relationship between age and the length of the follicular phase (percentage of cycle days at maximal tumescence). There also were positive relationships between age and the length of the estrous cycle, the percentage of cycle days at complete detumescence (i.e. amenorrhea), and the frequency of irregular cycles. Analysis of hormonal data revealed curvilinear relationships between age and both LH and FSH, with peaks above the menopausal threshold occurring around ages 35-40 years. Cycle and hormonal changes were similar to those in perimenopausal and menopausal women, providing evidence of perimenopause (at 30-35 years) and menopause (at 40 years) in the chimpanzee.

Effects of aging on hematology and serum clinical chemistry in chimpanzees (Pan troglodytes).

A number of age-related changes in physiological functions have been identified in macaques and humans. However, few studies have examined physiological aging in chimpanzees, despite the increasing age of the chimpanzee population. We documented age-related changes in seven hematology and 17 clinical chemistry parameters in 49 adult chimpanzees (17 males, 32 females) as a comparative viewpoint with human and macaque aging. Longitudinal data were analyzed using weighted linear and quadratic mixed effects regression models. Male chimpanzees exhibited a significant age-related increase in anemia risk, based on significant decreases in hemoglobin (F(1,49)=12.45, P=0.0009) and hematocrit (F(1,49)=15.42, P=0.0003). Both sexes exhibited significant age-related decreases in both kidney and liver function. Decreases in kidney function were noted by significant increases in blood urea nitrogen (F(1,45)=3.92, P=0.036) and creatinine (F(1,50)=5.63, P=0.022) as well as changes in electrolyte (i.e., sodium, potassium, phosphorous, chloride) balance. Declining liver function was based on significant increases in globulin (F(1,46)=32.34, P<0.0001) and decreases in albumin (F(1,48)=23.42, P<0.0001). These changes were most evident beginning at 25-30 years of age in males and 30-35 years of age in females. We recommend amending chimpanzee age classes to categorize males over 25 years and females over 30 years as aged.

Relationship between sunlight exposure, housing condition, and serum vitamin D and related physiologic biomarker levels in captive chimpanzees (Pan troglodytes).

In primates, the primary source of vitamin D is synthesis in the skin through sun exposure. Decreased sun exposure may lead to vitamin D deficiency and consequently other health issues. In laboratory, sanctuary, and zoo settings, chimpanzees (Pan troglodytes) may be housed indoors for prolonged periods of time without regular exposure to unfiltered sunlight. However, little research has examined the relationship between housing conditions and vitamin D serum levels in captive chimpanzees. In this study, we retrospectively compared serum levels of total vitamin D, calcium, ionic calcium, phosphorous, albumin, and alkaline phosphatase in 18 female and 12 male chimpanzees as they cycled between indoor-only and indoor-outdoor enclosures. Total vitamin D was significantly lower and alkaline phosphatase significantly higher when subjects were in the indoor-only enclosures compared with when they had regular access to outdoor enclosures. A vitamin D effect occurred only in young and prime-adult animals. Changes were significant in female but not in male chimpanzees. Calcium, ionic calcium, phosphorus, and albumin did not differ between indoor-only and indoor-outdoor enclosures. However, female chimpanzees exhibited significantly lower calcium and phosphorous levels while in the indoor-only enclosures. These results suggest that adult captive chimpanzees experience vitamin D deficiency when housed without regular access to unfiltered sunlight and that these effects may be more acute for adult female animals.

Functional evidence for differences in sperm competition in humans and chimpanzees.

Sperm competition occurs when the gametes of or more males compete for opportunities to fertilize a given set of ova. Previous studies have demonstrated that certain morphological characteristics are affected by sperm competition intensity (e.g. relative testes size and sperm midpiece volume). This study examined whether aspects of sperm energetics may also be affected by sexual selection. We compared the membrane potential of mitochondria in live sperm between H. sapiens (single partner mating system) and P. troglodytes (multiple partner mating system). Flow cytometry of sperm stained with the carbocyanine fluorescent dye JC-1 (an assay for mitochondrial membrane potential) revealed marked differences in red fluorescence intensity. P. troglodytes sperm showed significantly higher mitochondrial membrane potential. Mitochondria provide a substantial part of the energy required for sperm motility. A higher mitochondrial loading may therefore be associated with enhanced sperm motility and/or longevity. Additionally, examination of JC-1 red fluorescence levels before and after in vitro capacitation revealed further differences. Whereas chimpanzee sperm showed maintenance of membrane potential after capacitation (in some cases even an increase), sperm from humans consistently showed reduction in membrane potential. These results indicate that the sperm of human beings and chimpanzees exhibit marked differences in mitochondrial function, which are affected by selection pressures relating to sperm competition and that these pressures differ significantly between humans and chimpanzees.

Effects of two types and two genre of music on social behavior in captive chimpanzees (Pan troglodytes).

Is music just noise, and thus potentially harmful to laboratory animals, or can it have a beneficial effect? Research addressing this question has generated mixed results, perhaps because of the different types and styles of music used across various studies. The purpose of this study was to test the effects of 2 different types (vocal versus instrumental) and 2 genres (classical vocal versus 'easy-listening' vocal) of music on social behavior in 31 female and 26 male chimpanzees (Pan troglodytes). Results indicated that instrumental music was more effective at increasing affiliative behavior in both male and female chimpanzees, whereas vocal music was more effective at decreasing agonistic behavior. A comparison of 2 genre of vocal music indicated that easy-listening (slower tempo) vocal music was more effective at decreasing agonistic behavior in male chimpanzees than classical (faster tempo) vocal music. Agonistic behavior in females remained low (<0.5%) throughout the study and was unaffected by music. These results indicate that, like humans, captive chimpanzees react differently to various types and genres of music. The reactions varied depending on both the sex of the subject and the type of social behavior examined. Management programs should consider both type and genre when implementing a musical enrichment program for nonhuman primates.

Attempting to reduce regurgitation and reingestion in a captive chimpanzee through increased feeding opportunities: a case study.

Persistent regurgitation and reingestion is an abnormal behavior seen in many captive NHPs, particularly gorillas and chimpanzees. Successful reduction of regurgitation in captive gorillas has been achieved by providing browse, such as branches and leaves, and feeding continuously throughout the day. However, this modality has not been tested in chimpanzees. The authors tested these methods in a 15-year-old male chimpanzee (Pan troglodytes) with a seven-year history of regurgitation and reingestion by alternately providing additional opportunities to forage or giving daily browse. The results show that providing browse on a daily basis models the feeding behavior of wild chimpanzees and is a successful way to reduce regurgitation and reingestion in a chimpanzee unresponsive to previous dietary changes.

Development of guidelines for assessing obesity in captive chimpanzees (Pan troglodytes).

Many captive chimpanzees (Pan troglodytes) are subjectively considered to be overweight or obese. However, discussions of obesity in chimpanzees are rare in the literature, despite the acknowledged problem. No study to date has systematically examined obesity in captive chimpanzees. This project develops guidelines for defining obesity in captive chimpanzees through the examination of morphometric and physiologic characteristics in 37 adult female and 22 adult male chimpanzees. During each animal's biannual physical exam, morphometric data was collected including seven skinfolds (mm), body mass index (BMI), waist-to-hip ratio (WHR), and total body weight (kg). The morphometric characteristics were correlated with triglycerides and serum glucose concentration, to test the utility of morphometrics in predicting relative obesity in captive chimpanzees. Abdominal skinfold (triglyceride: F=3.83, P=0.05; glucose: F=3.83, P=0.05) and BMI (triglyceride: F=10.42, p=0.003; glucose: F=6.20, P=0.02) were predictive of increased triglycerides and serum glucose in females; however no morphometric characteristics were predictive of relative obesity in males. Results suggest that no males in this population are overweight or obese. For females, there were additional significant differences in morphometric (skinfolds, BMI, WHR, total body weight) and physiologic measurements (systolic and diastolic blood pressure, red blood cells) between individuals classified overweight and those classified non-overweight. Skinfold measurements, particularly abdominal, seem to be an accurate measure of obesity and thus potential cardiovascular risk in female chimpanzees, but not males. By establishing a baseline for estimated body fat composition in female captive chimpanzees, institutions can track individuals empirically determined to be obese, as well as obesity-related health problems. Zoo Biol 0:1-12, 2007. (c) 2007 Wiley-Liss, Inc.

The effects of aging on hormone and reproductive cycles in female chimpanzees (Pan troglodytes).

In contrast to those for human females, observational cycle data available for chimpanzees suggest that menstrual cycling, and thus reproductive potential, continues until near death. This study documents age-related changes in estrous cycling and hormone profiles in 14 female chimpanzees (Pan troglodytes) ranging in age from 31 to 50 y. Estrous data were analyzed from daily cycle charts, averaging 13.3 y of cycle data per subject, after omission of gestational and postpartum amenorrhea. Concentrations of total luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol (E2), and other hormones were assayed in serum samples taken biannually. Sample collection times were chosen to avoid the ovulatory LH and FSH peaks of the female's cycle and yielded a mean of 19.6 serum samples over an average of 14.4 y per subject. Analysis of cycle charts revealed a negative relationship between age and the percentage of cycle days at maximal tumescence. There also were positive relationships between age and the length of the estrous cycle and age and the percentage of cycle days at complete detumescence. Analysis of hormonal data revealed curvilinear relationships between age and both LH and FSH. These cycle and hormonal changes mirror those in perimenopausal and menopausal women. Our data provide evidence of perimenopause (at 30 to 35 y) and menopause (at 35 to 40 y) in the chimpanzee.

Controllability in environmental enrichment for captive chimpanzees (Pan troglodytes).

This study considers the use of nonsocial environmental enrichment by captive chimpanzees at the Primate Foundation of Arizona. The goal was to determine whether a relationship existed between controllability of enrichment items by captive chimpanzees and frequency of use. The study measured controllability, the ability of nonhuman animals to alter aspects of their environment by the potential destructibility of the enrichment item. This study examined additional factors that may affect enrichment use: individual age, sex, rearing history, social group composition, and availability of outdoor access. The chimpanzees in the study used destructible items--the enrichment category with the highest level of controllability--more than indestructible items across all age, sex, and rearing classes. Thus, controllability seems to be an important factor in chimpanzee enrichment. Younger individuals and groups with outdoor access used enrichment more than did older individuals and groups with indoor-only access. Individual sex, rearing history, and social group composition had minimal effects on enrichment use. These results support the importance of control to captive chimpanzees and further enable captive management to customize enrichment programs to the needs of particular animals.

Subspecies composition and founder contribution of the captive U.S. chimpanzee (Pan troglodytes) population.

Chimpanzees are presently classified into three subspecies: Pan troglodytes verus from west Africa, P.t. troglodytes from central Africa, and P.t. schweinfurthii from east Africa. A fourth subspecies (P.t. vellerosus), from Cameroon and northern Nigeria, has been proposed. These taxonomic designations are based on geographical origins and are reflected in sequence variation in the first hypervariable region (HVR-I) of the mtDNA D-loop. Although advances have been made in our understanding of chimpanzee phylogenetics, little has been known regarding the subspecies composition of captive chimpanzees. We sequenced part of the mtDNA HVR-I region in 218 African-born population founders and performed a phylogenetic analysis with previously characterized African sequences of known provenance to infer subspecies affiliations. Most founders were P.t. verus (95.0%), distantly followed by the troglodytes schweinfurthii clade (4.6%), and a single P.t. vellerosus (0.4%). Pedigree-based estimates of genomic representation in the descendant population revealed that troglodytes schweinfurthii founder representation was reduced in captivity, vellerosus representation increased due to prolific breeding by a single male, and reproductive variance resulted in uneven representation among male P.t.verus founders. No increase in mortality was evident from between-subspecies interbreeding, indicating a lack of outbreeding depression. Knowledge of subspecies and their genomic representation can form the basis for phylogenetically informed genetic management of extant chimpanzees to preserve rare genetic variation for research, conservation, or possible future breeding.

Neighbor effect: evidence of affiliative and agonistic social contagion in captive chimpanzees (Pan troglodytes).

Previous studies of captive chimpanzees have demonstrated the "neighbor effect," or social contagion, with respect to agonistic vocalizations and behaviors. The present study considers whether there is a relationship between behavior patterns in focal animals and the auditory signals of neighboring social groups. Using focal-group sampling, we collected 172.5 hr of data on 51 subjects (25 females and 26 males) housed in 10 social groups. We performed two-tailed Wilcoxon matched-pairs signed-rank tests to determine whether the relative frequency of the vocalizations (high vs. low) affected the behaviors. In keeping with past research, we found that agonistic noises and vocalizations from neighboring social groups had a significant effect on the rates of focal-group bluff displays, pant-hoots, and aggression (P<0.05). In addition, we also found significant relationships between grooming behavior and vocalizations in focal groups, and grooming vocalizations from neighboring groups (P<0.05). The results suggest that social contagion is not limited to aggressive behaviors, but also occurs for affiliative behavior patterns.

Training captive chimpanzees to cooperate for an anesthetic injection.

Captive animals trained to cooperate with routine medical procedures, such as injections, may experience less aggression and anxiety than those forced to comply through the use of restraints. The authors used positive reinforcement training to teach captive chimpanzees to present a body part for anesthetic injection and determined the time investment necessary for initial training and duration of maintenance of the behavior after completion of the training.

Normal hematologic and serum clinical chemistry values for captive chimpanzees (Pan troglodytes).

In the study reported here, reference intervals for hematologic and serum clinical chemistry variables in the chimpanzee (Pan troglodytes) were developed and characterized. Data were collected longitudinally across a 10-year period for 86 subjects at the Primate Foundation of Arizona (PFA). Variables included nine standard hematologic and 25 standard serum clinical chemistry values. An analysis of variance (ANOVA) was used to test for main effects by age and sex. In addition, PFA mean and range values were compared with those published for humans and six other chimpanzee colonies. The ANOVA results suggest an age effect on hematologic (mean corpuscular hemoglobin, mean corpuscular volume, neutrophils) and serum clinical chemical (creatinine, total protein, globulin, tryglycerides, direct bilirubin, iron, (gamma-glutamyltransferase, alanine transaminase, creatine kinase) values. In addition, sex had a main effect on several variables (red blood cells, hemoglobin concentration, hematocrit, uric acid and sodium concentrations, and aspartate transminase and creatine kinase activities); values for males were greater than those for females. Further, human and chimpanzee mean and range values often were indistinguishable from one another. However, changes in human and chimpanzee values associated with age differ and suggest that hematologic and serum clinical chemistry values may be differentially affected by physical and sexual maturation in humans and chimpanzees.

A stereo music system as environmental enrichment for captive chimpanzees.

Music has been shown to have beneficial effects on humans but little is known about the effects of music on nonhuman primates in biomedical research settings. The authors monitored the effects of music on the behavior of captive chimpanzees and found that music had significant positive effects, including a reduction in agitated and aggressive behaviors.

Weight gain in captive chimpanzee infants: Comparisons by sex, rearing, and colony.

Weight gain has been monitored for 13 years in a mixed longitudinal study of captive chimpanzee growth and development. This report presents results of a comparative analysis of weight relative to age in 175 animals during the first 24 months in four sex/rearing groups (hand-reared females, hand-reared males, mother-reared females, and mother-reared males) from three colonies with different physical, nutritional, and social environments (Primate Foundation of Arizona, University of Texas M.D. Anderson Cancer Center Department of Veterinary Resources, Bastrop, TX, and White Sands Research Center, Alamagordo, NM). The Lowess method is used to generate fits of weight vs. age for each group and colony, with which individual animals at these and other colonies may be compared for assessment of developmental status. Comparisons of the curves, using the jackknife approach, show that there are significant differences between the curves, indicating that rearing and environmental parameters may be factors in weight gain rate and must be considered in such an assessment. Rearing effects may be the dominant of these factors in weight gain. © 1996 Wiley-Liss, Inc.