Sex and Urbanicity Contribute to Variation in Lymphocyte Distribution across Ugandan Populations

Management of patient care and interpretation of research data require evaluation of laboratory results in the context of reference data from populations with known health status to adequately diagnose disease or make a physiological assessment. Few studies have addressed the diversity of lymphocyte subsets in rural and urban Ugandan populations. Here, 663 healthy blood bank donors from semi-urban centers of Kampala consented to participate in a study to define lymphocyte reference ranges. Whole blood immunophenotyping was performed to determine the frequency and absolute counts of T, B, and NK cells using clinical flow cytometry. Results from blood bank donors were compared to a rural cohort from the district of Kayunga and more urban clinical trial participants from the capital city, Kampala. Relationships between hematological and lymphocyte parameters were also explored. In the semi-urban blood donors, females were significantly different from males in all parameters except the frequency of CD8 T and B cells. Females had higher absolute counts of CD4 T, CD8 T and B cells, whereas males had higher NK cell counts. NK cell frequency and counts were significantly higher in semi-urban blood donors, regardless of sex, compared to more urban study participants. CD8 T cell frequency and counts were significantly higher in the blood donors compared to the rural participants, irrespective of sex. Interestingly, basophil counts were positively associated with overall T cell counts. These findings suggest that both sex and level of cohort urbanicity may influence lymphocyte subset distributions in Ugandans.     

Reference ranges for the clinical laboratory derived from a rural population in Kericho, Kenya

The conduct of Phase I/II HIV vaccine trials internationally necessitates the development of region-specific clinical reference ranges for trial enrollment and participant monitoring. A population based cohort of adults in Kericho, Kenya, a potential vaccine trial site, allowed development of clinical laboratory reference ranges. Lymphocyte immunophenotyping was performed on 1293 HIV seronegative study participants. Hematology and clinical chemistry were performed on up to 1541 cohort enrollees. The ratio of males to females was 1.9:1. Means, medians and 95% reference ranges were calculated and compared with those from other nations. The median CD4+ T cell count for the group was 810 cells/microl. There were significant gender differences for both red and white blood cell parameters. Kenyan subjects had lower median hemoglobin concentrations (9.5 g/dL; range 6.7-11.1) and neutrophil counts (1850 cells/microl; range 914-4715) compared to North Americans. Kenyan clinical chemistry reference ranges were comparable to those from the USA, with the exception of the upper limits for bilirubin and blood urea nitrogen, which were 2.3-fold higher and 1.5-fold lower, respectively. This study is the first to assess clinical reference ranges for a highland community in Kenya and highlights the need to define clinical laboratory ranges from the national community not only for clinical research but also care and treatment.     

Cotton-top tamarins (Saguinus oedipus oedipus): Hematologic reference values and hemopathologic responses

Hematologic reference values have been established for captive adult cotton-top tamarins (Saguinus oedipus oedipus) by carrying out full blood counts and fibrinogen estimation on 43 clinically normal animals. Females were shown to have significantly lower hemoglobin levels, red cell counts and packed cell volumes, and higher reticulocyte counts than males. The reference values were used to identify abnormal changes in the blood of 13 clinical cases. Marked neutrophilia was found in animals with localized bacterial infections, and a degenerative left shift was recorded in an individual with streptococcal septicemia. Three cases of unexplained progressive muscle wasting showed Heinz body anemia and abnormal white cell changes. These hematologic responses have been compared where possible with those recorded in other species of Callithricidae.     

Reference Ranges of Cholesterol Sub-Fractions in Random Healthy Adults in Ouagadougou,Burkina Faso

In Burkina Faso, the values that serve as clinical chemistry reference ranges are those provided by European manufacturers’ insert sheets based on reference of the Western population. However, studies conducted so far in some African countries reported significant differences in normal laboratory ranges compared with those of the industrialized world. The aim of this study was to determine reference values of cholesterol fractions in apparently normal adults in Burkina Faso that could be used to better assess the risks related to cardiovascular diseases. Study population was 279 healthy subjects aged from 15 to 50 years including 139 men and 140 women recruited at the Regional Center of Blood Transfusion of Ouagadougou, capital city of Burkina Faso (West Africa). Exclusion criteria based on history and clinical examination were used for defining reference individuals. The dual-step precipitation of HDL cholesterol sub-fractions using dextran sulfate was performed according to the procedure described by Hirano. The medians were calculated and reference values were determined at 2.5^th and 97.5^th percentiles. The median and upper ranges for total cholesterol, LDL cholesterol, total HDL cholesterol and HDL2 cholesterol were observed to be higher in women in comparison to men (p <0.05). These reference ranges were similar to those derived from other African countries but lower than those recorded in France and in USA. This underscores the need for such comprehensible establishment of reference values for limited resources countries. Our study provides the first cholesterol sub-fractions (HDL2 and HDL3) reference ranges for interpretation of laboratory results for cardiovascular risk management in Burkina Faso.     

Cerebrospinal fluid glucose and lactate: age-specific reference values and implications for clinical practice

Cerebrospinal fluid (CSF) analysis is an important tool in the diagnostic work-up of many neurological disorders, but reference ranges for CSF glucose, CSF/plasma glucose ratio and CSF lactate based on studies with large numbers of CSF samples are not available. Our aim was to define age-specific reference values. In 1993 The Nijmegen Observational CSF Study was started. Results of all CSF samples that were analyzed between 1993 and 2008 at our laboratory were systematically collected and stored in our computerized database. After exclusion of CSF samples with an unknown or elevated erythrocyte count, an elevated leucocyte count, elevated concentrations of bilirubin, free hemoglobin, or total protein 9,036 CSF samples were further studied for CSF glucose (n = 8,871), CSF/plasma glucose ratio (n = 4,516) and CSF lactate values (n = 7,614). CSF glucose, CSF/plasma glucose ratio and CSF lactate were age-, but not sex dependent. Age-specific reference ranges were defined as 5–95^th percentile ranges. CSF glucose 5^th percentile values ranged from 1.8 to 2.9 mmol/L and 95^th percentile values from 3.8 to 5.6 mmol/L. CSF/plasma glucose ratio 5^th percentile values ranged from 0.41 to 0.53 and 95^th percentile values from 0.82 to 1.19. CSF lactate 5^th percentile values ranged from 0.88 to 1.41 mmol/L and 95^th percentile values from 2.00 to 2.71 mmol/L. Reference ranges for all three parameters were widest in neonates and narrowest in toddlers, with lower and upper limits increasing with age. These reference values allow a reliable interpretation of CSF results in everyday clinical practice. Furthermore, hypoglycemia was associated with an increased CSF/plasma glucose ratio, whereas hyperglycemia did not affect the CSF/plasma glucose ratio.     

Reference Values for Clinical Laboratory Parameters in Young Adults in Maputo,Mozambique

Background

Clinical laboratory reference values from North American and European populations are currently used in most Africans countries due to the absence of locally derived reference ranges, despite previous studies reporting significant differences between populations. Our aim was to define reference ranges for both genders in 18 to 24 year-old Mozambicans in preparation for clinical vaccine trials.

Methods

A cross-sectional study including 257 volunteers (102 males and 155 females) between 18 and 24 years was performedat a youth clinic in Maputo, Mozambique. All volunteers were clinically healthy and human immunodeficiency virus, Hepatitis B virus and syphilis negative.Median and 95% reference ranges were calculated for immunological, hematological and chemistry parameters. Ranges were compared with those reported based on populations in other African countries and the US. The impact of applying US NIH Division of AIDS (DAIDS) toxicity tables was assessed.

Results

The immunology ranges were comparable to those reported for the US and western Kenya.There were significant gender differences in CD4⁺ T cell values 713 cells/µL in males versus 824 cells/µL in females (p<0.0001). Hematologic values differed from the US values but were similar to reports of populations in western Kenya and Uganda. The lower and upper limits of the ranges for hemoglobin, hematocrit, red blood cells, white blood cells and lymphocytes were somewhat lower than those from these African countries. The chemistry values were comparable to US values, with few exceptions. The upper limits for ALT, AST, bilirubin, cholesterol and triglycerides were higher than those from the US. DAIDStables for adverse events predicted 297 adverse events and 159 (62%) of the volunteers would have been excluded.

Conclusion

This study is the first to determine normal laboratory parameters in Mozambique. Our results underscore the necessity of establishing region-specific clinical reference ranges for proper patient management and safe conduct of clinical trials.     

Laboratory and Field Evaluation of the Partec CyFlow MiniPOC for Absolute and Relative CD4 T-Cell Enumeration

BackgroundA new CD4 point-of-care instrument, the CyFlow miniPOC, which provides absolute and percentage CD4 T-cells, used for screening and monitoring of HIV-infected patients in resource-limited settings, was introduced recently. We assessed the performance of this novel instrument in a reference laboratory and in a field setting in Senegal.MethodologyA total of 321 blood samples were obtained from 297 adults and 24 children, all HIV-patients attending university hospitals in Dakar, or health centers in Ziguinchor. Samples were analyzed in parallel on CyFlow miniPOC, FACSCount CD4 and FACSCalibur to assess CyFlow miniPOC precision and accuracy.ResultsAt the reference lab, CyFlow miniPOC, compared to FACSCalibur, showed an absolute mean bias of -12.6 cells/mm³ and a corresponding relative mean bias of -2.3% for absolute CD4 counts. For CD4 percentages, the absolute mean bias was -0.1%. Compared to FACSCount CD4, the absolute and relative mean biases were -31.2 cells/mm³ and -4.7%, respectively, for CD4 counts, whereas the absolute mean bias for CD4 percentages was 1.3%. The CyFlow miniPOC was able to classify HIV-patients eligible for ART with a sensitivity of ≥ 95% at the different ART-initiation thresholds (200, 350 and 500 CD4 cells/mm3). In the field lab, the room temperature ranged from 30 to 35°C during the working hours. At those temperatures, the CyFlow miniPOC, compared to FACSCount CD4, had an absolute and relative mean bias of 7.6 cells/mm³ and 2.8%, respectively, for absolute CD4 counts, and an absolute mean bias of 0.4% for CD4 percentages. The CyFlow miniPOC showed sensitivity equal or greater than 94%.ConclusionThe CyFlow miniPOC showed high agreement with FACSCalibur and FACSCount CD4. The CyFlow miniPOC provides both reliable absolute CD4 counts and CD4 percentages even under the field conditions, and is suitable for monitoring HIV-infected patients in resource-limited settings.     

Fertilization and male fertility in the rotifer Brachionus plicatilis

The role of males in fertilization in the rotifer Brachionus plicatilis was examined. Neonate mictic females sometimes escaped fertilization even when sperm were present in their pseudocoeloms. Males made a major contribution to this fertilization failure through temporary infertility. As males aged, sperm numbers and motility decreased. Maternal diet was found to play a primary role in determining the fertilizing capacity of the F₁ males. Females from log phase populations produced males with higher sperm counts and motility than females from stationary phase populations. A conditioning factor in the medium may increase sperm number and fertilization rate.     

Reproductive behavior of largemouth triplefin Ucla xenogrammus (Tripterygiidae) on reefs of Kuchierabu-jima Island, southern Japan

Reproductive behaviors of the largemouth triplefin Ucla xenogrammus were observed on reefs of Kuchierabu-jima Island, southern Japan. Males exclusively maintained home ranges including spawning sites on vertical walls of overhanging reefs where filamentous algae were densely distributed. Females visited a male's home range to release adhesive eggs into the dense algae. Males guarded multiple egg clutches, but rarely showed fanning behaviors or mouth cleaning to the eggs. Aquarium experiments showed that guarding males had no significant effect on egg development. Because of the favorable water exchange conditions around their spawning sites, male U. xenogrammus could avoid the energy costs for egg care.     

Serum Reference Values for Leptin in Healthy Infants

ObjectiveReports on leptin concentrations in pediatric populations lack reference values for infants in the first months of life. Our study was conducted on healthy full-term infants between 2002 and 2012 to determine serum leptin reference values in subjects less than 18 months old.MethodsRoutine outpatient blood tests for serum leptin were performed on 317 infants using a radioimmunoassay method. The median and 10^th–90^th percentiles were calculated to obtain reference values using quantile regression. Values established in this study were compared with another independent cohort of 110 infants.ResultsThe median (IQR) serum leptin concentration in the infants was 2.37 (3.26) ng/ml (n = 317). The median leptin concentration was 2.81 (3.49) ng/ml (n = 202) in infants younger than 6 months of age, 1.44 (2.27) ng/ml (n = 59) in infants between 6–12 months of age and 1.77 (2.05) ng/ml (n = 56) in infants between 12–18 months of age. We obtained leptin reference values based on age by estimating the lower and upper percentiles. In the entire cohort, the median (IQR) leptin concentration was 2.22 (3.11) ng/ml in males (n = 168) and 2.60 (3.32) ng/ml in females (n = 149). According to the type of feeding median serum leptin concentration was higher in breast-fed infants (n = 188) than in formula-fed infants (n = 129) (2.63 (3.34) ng/ml vs. 2.12 (2.77) ng/ml; p<0.05).ConclusionsOur data revealed no gender difference in leptin concentration in early infancy. After 6 months of life, leptin concentrations decreased slightly. We used a large cohort to confirm that breast-fed infants had significantly higher serum leptin levels than formula-fed infants during the first 6 months of life, although this difference disappeared later in life. In this study, we defined the leptin reference range in healthy infants in the first 18 months of life according to the Clinical and Laboratory Standards Institute (CLSI).     

A comparison of chiasma frequency and distribution between sexes in three species of grasshoppers

Chiasma frequency and distribution have been compared in both sexes of the grasshoppers Chortoicetes terminifera (n=11+X) (Oedipodinae) Parapleurus alliaceus (n=11+X) (Oedipodinae) and Chrysochraon dispar (n=8+X) (Gomphocerinae). In the last two species, chiasmata in the males are terminally localised in all except the shortest bivalents. Strict chiasma localisation was not found in the females but chiasmata were rare near to the centromere. The sexes had different autosomal mean chiasma frequencies. Cell means were: C. terminifera Males 13.07, Females 13.02; P. alliaceus Males 12.32, Females 14.47; C. dispar Males 12.56, Females 13.57. Possible mechanisms affecting the distribution of chiasmata, and the significance of the intersex and interspecies difference, are discussed.     

Polygamous mating system and protogynous sex change in the gobiid fish <Emphasis Type="Italic">Fusigobius neophytus</Emphasis>

Whereas mating behaviors and social structure have been studied extensively in monogamous hermaphroditic gobiid species, such studies are relatively limited for polygamous gobiid species. To investigate the reproductive strategy of polygamous gobies, mating groups of the common fusegoby Fusigobius neophytus were observed on reefs of Kuchierabu-jima Island, southern Japan. Males established mating nests on flat-rock surfaces within their territorial home ranges on sandy rubble flats. Females maintained independent home ranges outside the male home ranges during nonreproductive periods, but they shifted their home ranges to overlap with male ranges and actively visited male mating nests during their reproductive periods (1–3 days at ca. 7-day intervals). Females often changed mates during their serial mating. The mating system used by the common fusegoby fits with the definition of male-territory-visiting polygamy. The sex ratio within the study population was female-biased. Nest-holding males were significantly larger than females and were polygynous (mating with up to eight females). These characteristics fit well with the prediction of protogyny by the size-advantage model. Some of the females were observed to undergo functional sex changes to nest-holding males. In addition, small floating males demonstrated sneaking behavior. None of the floating males were derived from females that had changed sex, suggesting a diandric life-history pathway for F. neophytus.     

Parental differences in brood provisioning by Hen Harriers Circus cyaneus

Capsule Females varied their provisioning patterns according to brood age and brood size, whereas males did not.

Aims To quantify how parents balance the needs of their offspring for food and protection.

Methods We studied 13 nests from hides and spent on average 101 hours per nest monitoring prey types, provisioning rate and the time spent at the nest by both sexes in relation to brood size and brood age.

Results Males always provided more food than females. Males brought similar amounts of prey items irrespective of brood size and nestling age, whereas females brought more prey and bigger items to larger and older broods. Females spent less time brooding larger broods, particularly early on.

Conclusions Hen Harrier parents share the provisioning burden, with each parent delivering prey as a function of brood care requirements, hunting capability and the behaviour of the other parent.     

Male Corncrakes Crex crex extend their home ranges by visiting the territories of neighbouring males

Capsule Radiotracked male Corncrake often intruded on the territories of neighbouring males.

Aims To test that intruders' visits are goal-directed, not just a by-product of extended spatial activity during daylight hours.

Methods Using radiotelemetry, we sampled a total of 20 three-day home ranges from 11 tagged males. We recorded daily vocal activity and used a permutation test to see if the movements of tracked males were independent of the position of neighbouring males.

Results The majority of males who had a neighbouring male, up to approximately 600 m from their night calling site, undertook goal-directed visits to the neighbour's territory. Males undertook these visits every day, or every other day, when the neighbours were close. Males undertook visits approximately once every three days when they were more distant. The time spent in the neighbour's territory was longest where the distance between night calling sites was about 200 m. Males tended to be silent in neighbour's territory, apparently to prevent confrontation. Otherwise the distance of neighbouring males did not significantly affect daytime vocal activity. Visiting males tended to sing more often in their home territories.

Conclusions Daily movement of the majority of males was towards the neighbouring male's calling site. We suggest that the purpose of these visits was to seek females. These males may try to drive a female into their territory or gain extra-pair copulation.     

CLSI-Derived Hematology and Biochemistry Reference Intervals for Healthy Adults in Eastern and Southern Africa

Background

Clinical laboratory reference intervals have not been established in many African countries, and non-local intervals are commonly used in clinical trials to screen and monitor adverse events (AEs) among African participants. Using laboratory reference intervals derived from other populations excludes potential trial volunteers in Africa and makes AE assessment challenging. The objective of this study was to establish clinical laboratory reference intervals for 25 hematology, immunology and biochemistry values among healthy African adults typical of those who might join a clinical trial.

Methods and Findings

Equal proportions of men and women were invited to participate in a cross sectional study at seven clinical centers (Kigali, Rwanda; Masaka and Entebbe, Uganda; two in Nairobi and one in Kilifi, Kenya; and Lusaka, Zambia). All laboratories used hematology, immunology and biochemistry analyzers validated by an independent clinical laboratory. Clinical and Laboratory Standards Institute guidelines were followed to create study consensus intervals. For comparison, AE grading criteria published by the U.S. National Institute of Allergy and Infectious Diseases Division of AIDS (DAIDS) and other U.S. reference intervals were used. 2,990 potential volunteers were screened, and 2,105 (1,083 men and 1,022 women) were included in the analysis. While some significant gender and regional differences were observed, creating consensus African study intervals from the complete data was possible for 18 of the 25 analytes. Compared to reference intervals from the U.S., we found lower hematocrit and hemoglobin levels, particularly among women, lower white blood cell and neutrophil counts, and lower amylase. Both genders had elevated eosinophil counts, immunoglobulin G, total and direct bilirubin, lactate dehydrogenase and creatine phosphokinase, the latter being more pronounced among women. When graded against U.S.-derived DAIDS AE grading criteria, we observed 774 (35.3%) volunteers with grade one or higher results; 314 (14.9%) had elevated total bilirubin, and 201 (9.6%) had low neutrophil counts. These otherwise healthy volunteers would be excluded or would require special exemption to participate in many clinical trials.

Conclusions

To accelerate clinical trials in Africa, and to improve their scientific validity, locally appropriate reference ranges should be used. This study provides ranges that will inform inclusion criteria and evaluation of adverse events for studies in these regions of Africa.     

Hematologic and serum chemistry values of the captive European wildcat

Hematologic and serum chemistry values were determined for 20 adult captive European wildcats (Felis silvestris) in Lleida (Catalonia, Spain). Seven wildcats (4 females and 3 males) were captured in the wild and 13 (4 females and 9 males) were born and raised in captivity. Samples were collected between September and December from 1993 to 1998. Blood was obtained by jugular venipuncture after administration of either ketamine and xylazine or ketamine and medetomidine. Females had significantly higher mean eosinophil counts, albumin concentration, (and A/G ratio) and lower mature neutrophil counts, although these differences were not clinically relevant. Results for many of the blood parameters fall within the reference range for domestic cats. Mean values for alanine aminotransferase, aspartate aminotransferase, creatine phosphokinase, and lactate dehydrogenase activities as well as blood urea nitrogen, glucose, and sodium concentrations were higher than the upper limit of the reference range for domestic cats.     

Navigation by Male Crab Spiders <Emphasis Type="Italic">Misumenoides formosipes</Emphasis> (Araneae: Thomisidae): Floral Cues May Aid in Locating Potential Mates

The crab spider Misumenoides formosipes is an ambush predator whose males search for relatively sedentary females within a heterogeneous habitat. Females are receptive to mating immediately after their adult molt and a male biased adult sex ratio together with precopulatory guarding places a premium on male ability to locate females quickly. It is unknown what cues males use to find females; we report here on male movements in association with floral cues. Males in field trials moved towards inflorescences more frequently when both visual and chemical cues were available, than when chemical cues were eliminated. Males in lab trials chose an inflorescence over leaf substrates even in the absence of visual cues. These findings support the hypothesis that M. formosipes males could utilize floral chemistry as a navigational cue in mate searches.     

Poor acute outcome in congestive heart failure is associated with increases in the plasma static oxidation–reduction potentials (sORP) in men but not in women

Objectives: In congestive heart failure (CHF), men are younger, more likely to have reduced ejection fraction (HF-rEF), and to be diabetic compared to women. Despite this, sex differences in oxidative stress have yet to be explored in CHF.

Methods: Data from 67 males and 63 females hospitalized for CHF were collected. Static oxidation–reduction potential (sORP), a relative indicator of oxidative stress, and capacity ORP (icORP), a relative indicator of antioxidant capacity, were measured from plasma samples. We examined whether sex modified the relationship between ORP and hospital discharge disposition (poor outcome: death, hospice), along with other demographics, medications, and diagnostic parameters.

Results: Males with poor outcomes had higher sORP and icORP values than females (P?P?>?0.05). Males were younger and more likely to have HF-rEF and diabetes. Controlling for these variables did not account for the sex differences in ORP measures. Regardless of sex, higher creatinine was related to higher sORP and icORP, while lower magnesium and potassium were related to higher sORP and icORP, respectively.

Discussion: Increases in sORP during CHF are partially affected by sex and acute outcomes, but are also related to variables without sexual biases.     

Inflammatory markers in end-stage renal disease patients on haemodialysis

BackgroundCXC chemokine ligand 16 (CXCL16) is an inflammatory chemokine that mediates renal infiltration of macrophages and activated T cells. Aim: To investigate serum levels of CXCL16 in patients undergoing hemodialysis and their correlation with other inflammatory markers such as C-reactive protein (CRP) and intact parathyroid hormone (iPTH).MethodsThe study included 40 hemodialysis patients (22 males) and 40 age and gender-matched controls (24 males). Fasting blood sugar (FBS), urea, creatinine, calcium and inorganic phosphorous were assayed in participants using routine methods, glycosylated hemoglobin (HbA1c) by quantitative chromatographic spectrophotometry, iPTH by chemiluminescent microparticle immunoassay, CRP by nephelometry and CXCL16 by ELISA technique.ResultsSerum CXCL16, CRP, PTH, FBS, HbA1c, phosphorus, urea, and creatinine levels were significantly higher in hemodialysis patients compared to controls (p<0.00001). No statistically significant differences were observed between patients and controls for calcium. Serum CXCL16 levels correlated positively with CRP (r=0.956, p<0.00001) and iPTH (r=-0.403, p<0.001). Hemodialysis patients (diabetics or hypertensives) had significantly higher CXCL16 levels compared to non-diabetics or non-hypertensives.ConclusionsHigh levels of serum CXCL16, CRP and iPTH reflect the inflammatory status of hemodialysis patients and help avoid complications. Serum CXCL16 could be used as a biomarker together with CRP in these patients.     

Hematological and serum chemistry profiles of free-ranging southern two-toed sloths in French Guiana.

Free-ranging southern two-toed sloths (Choloepus didactylus) were translocated during the flooding of a forest at a hydroelectric dam site in French Guiana. Over an 11 mo period blood samples were collected from 90 sloths (38 males, 52 females) in order to determine hematological and serum chemistry reference values. Mean values and range of values were calculated for 13 hematological and 21 serum chemistry parameters. Variations associated with sex, age and reproductive status were identified. Males had a significantly lower red blood cell count than females. Immature animals had more monocytes while adults had more neutrophils and higher mean corpuscular hemoglobin concentration. Aspartate aminotransferase and triglyceride values were higher in young than in adult sloths but uric acid was lower. Lactating females showed lower red blood cells count and iron levels than non-lactating females. These profiles will help to provide reliable baseline data for medical evaluation of sloths.