Determining Hematological Reference Intervals of the Blood Count in a Healthy Moroccan Adult Population (Al Hoceima Province).

BACKGROUND: The hemogram is the most frequently prescribed laboratory test. It guides the complementary tests essential to the diagnosis and follow-up of the patient. Hematology reference values can be influenced by several parameters such as environmental and genetic factors, rendering it essential to define reference intervals (RIs) for specific populations. This study aimed to determine RIs from a population of healthy adults in Al Hoceima province by following the procedures recommended by the IFCC-CLSI guidelines in 2008 and comparing them to those of similar studies conducted in various countries. METHODS: We initially recruited 977 healthy adults from 18 to 60 years old including 255 men and 722 women, who presented themselves at the provincial hospital center of Al Hoceima (Morocco) for health checkups from February 2019 to September 2019. The erythrocyte, leukocyte, and platelet parameters were investigated using hematology analyzer ABX Pentra XL80 HORIBA® (HORIBA ABX SAS, Kyoto, Japan). RESULTS: The results showed that the RIs of the blood count parameters searched were more or less close to the RIs determined in Caucasian populations. Nevertheless, they were low compared to the limits of the RIs published in the literature for certain erythrocyte and platelet parameters. Our RIs were also similar in their majority, except for some parameters, to the RIs published very recently in a study conducted on a Moroccan population in the Tangier-Tetouan region. In addition, our intervals differ in their majority from those published in a study of a Ghanaian population. CONCLUSIONS: The differences reported by this preliminary work reinforce the need to establish hemogram RIs specific to the Moroccan population through more extensive studies carried out in different regions of Morocco. These studies should avoid misdiagnosis and allow physicians to interpret hematological tests more specifically.

Reference Intervals for Routine Biochemical Markers and Hematological Indices Derived from Healthy Adults in the Mediterranean Region of Morocco.

BACKGROUND: Reference intervals (RIs) for biochemical and hematological parameters are fundamental tools for clinical diagnosis, management, and therapeutic follow-up. In Morocco, the RIs used by clinical laboratories and physicians are derived from western populations. Also, RIs of biochemical and hematological parameters specific to the various Moroccan areas are lacking. This study aimed to determine RIs for biochemical and hematological parameters in apparently healthy voluntary adults by following the procedures recommended by the IFCC-CLSI guidelines in 2008 and comparing them to those of literature and other countries. METHODS: A total of 768 healthy adults from 18 to 60 years old were recruited. Complete blood count and biochemical analyses were performed using hematology analyzer Sysmex KX21N® (Sysmex Corporation, Kobe, Japan) and COBAS INTEGRA®400 plus biochemistry analyzer (Roche, Diagnostics GmbH, Germany) at the laboratory of the hospital Mohamed VI of M'diq, Morocco, and went into effect between November 2017 and December 2020. The data analysis was made by the software SPSS 20.0 and RIs have been established by using the 2.5th and 97.5th percentiles. RESULTS: RIs established include: glucose 3.90 - 6.76 mmol/L for males and 4.01 - 6.87 mmol/L for females; alanine aminotransferase 5.60 - 40.07 U/L for males and 5.60 - 38.71 U/L for females; aspartate aminotransferase 5.60 - 40.08 for males and 5.89 - 39.90 U/L for females; creatinine 47.73 - 113.15 µmol/L for males and 44.64 - 102.28 µmol/L for females; urea 2.2 - 7.6 mmol/L for males and 1.90 - 7.5 mmol/L for females; total cholesterol 2.71 - 5.46 mmol/L for males and 2.64 - 5.89 mmol/L for females; triglycerides 0.58 - 2.01 mmol/L for males and 0.55 - 2.08 mmol/L for females; high-density lipoprotein cholesterol 1.40 - 1.50 mmol/L for males and 1.40 - 1.65 mmol/L for females; and uric acid 157.3 - 410.8 µmol/L for males and 146.1 - 388.5 µmol/L for females. Concerning the hematological parameters, a significant difference (p < 0.05) between both genders was noted for the majority of pa rameters. CONCLUSIONS: The present study underlines the importance to establish RIs specific to the Moroccan population in each region for a more rational and reliable interpretation of biochemical and hematological testing in order to avoid errors in diagnosis and treatment of patients.

Pinealectomy and gonadectomy modulate amplitude, but not photoperiodic modulation of Clock gene expression in the Syrian hamster suprachiasmatic nuclei.

The duration of daytime light phase (photoperiod) controls reproduction in seasonal mammals. Syrian hamsters are sexually active when exposed to long photoperiod, while gonadal atrophy is observed after exposure to short photoperiod. The photorefractory period, or photorefractoriness, is a particular state of spontaneous recrudescence of sexual activity that occurs after a long-term exposure to short photoperiod. Expression of core clock genes in the master circadian clock contained in the suprachiasmatic nuclei depends on photoperiodic conditions. Interestingly, the expression of the Clock gene is also modified in photorefractory Syrian hamsters. Since melatonin and testosterone levels in seasonal species are dependent on photoperiod, photoperiodic variations of Clock mRNA levels in the suprachiasmatic clock could be a consequence of these hormonal changes. To test this hypothesis, we analysed the effects of pinealectomy on Clock mRNA changes due to long to short photoperiod transition and of gonadectomy on Clock mRNA levels in photorefractory period. Our data show that the suprachiasmatic integration of the short photoperiod (assessed by a rhythmic expression profile of Clock) is independent of the presence of melatonin. Furthermore, constitutively low expression of Clock observed during the photorefractory period does not require the presence of either melatonin or testosterone. However, we show that both hormones provide positive feedback on average levels of Clock expression. Thus, our data support the hypothesis that daily variations of Clock levels in the suprachiasmatic nuclei are influenced by photoperiodic changes and the time spent in short photoperiod, independently of seasonal modifications of melatonin or testosterone levels.

Out of Africa: demographic and colonization history of the Algerian mouse (Mus spretus Lataste).

North Africa is now recognized as a major area for the emergence and dispersal of anatomically modern humans from at least 315 kya. The Mediterranean Basin is thus particularly suited to study the role of climate versus human-mediated changes on the evolutionary history of species. The Algerian mouse (Mus spretus Lataste) is an endemic species from this basin, with its distribution restricted to North Africa (from Libya to Morocco), Iberian Peninsula and South of France. A rich paleontological record of M. spretus exists in North Africa, suggesting hypotheses concerning colonization pathways, and the demographic and morphologic history of this species. Here we combined genetic (3 mitochondrial DNA loci and 18 microsatellites) and climatic niche modeling data to infer the evolutionary history of the Algerian mouse. We collected 646 new individuals in 51 localities. Our results are consistent with an anthropogenic translocation of the Algerian mouse from North Africa to the Iberian Peninsula via Neolithic navigators, probably from the Tingitane Peninsula. Once arrived in Spain, suitable climatic conditions would then have favored the dispersion of the Algerian mice to France. The morphological differentiation observed between Spanish, French and North African populations could be explained by a founder effect and possibly local adaptation. This article helps to better understand the role of climate versus human-mediated changes on the evolutionary history of mammal species in the Mediterranean Basin.

Hematological parameters of the blood count in a healthy population of pregnant women in the Northwest of Morocco (Tetouan-M'diq-Fnideq provinces).

INTRODUCTION: Numerous biological parameters are physiologically modified during normal pregnancy, in particular hematology. The knowledge of these modifications of the maternal body by biologists and clinicians allows the screening of possible anomalies. In Morocco, the reference values of the complete blood count test for pregnant woman are missing, as are those specific to different trimesters of pregnancy. The aim of this study is to look for the reference values for healthy pregnant women of the Northwest region of Morocco, to compare them to those of non-pregnant women (control) and to those of the literature. METHODS: Blood samples were taken voluntarily from 3898 healthy pregnant women from 18 to 46 years old who presented themselves at the center of health Kalaa and at the service of gynecology obstetrics of the Provincial Hospital Center of M'diq (Morocco), for prenatal care. To establish the reference intervals of the CBC for non-pregnant women, a control group was constituted by 7035 healthy women from 18 to 50 years old selected according to the Moroccan law of blood donation. The CBC was measured on a Sysmex KX21N® analyzer. For each sample a systematic blood smear was done to determine the leukocyte differential. RESULTS: A statistically significant difference between the pregnant women and control group was noted (p < 0.05) for all the hematological parameters: red blood cells, hematocrit, hemoglobin, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, leukocytes, neutrophils, basophils, eosinophils, lymphocytes, monocytes, platelets and mean platelet volume. So, the comparison of the averages established between the first, second and third trimester of pregnancy showed the existence of a significant variation with regard to all the parameters of the CBC test looked for (p < 0.001). CONCLUSION: The present study provides additional baseline data for basic hematological parameters in healthy pregnant Moroccan women and concluded that pregnancy in women has the tendency to alter some hematological indices. For these reasons, there is an interest to take these modifications into account for optimal maternal and fetal medical care.

Establishment of the hematology reference intervals in a healthy population of adults in the Northwest of Morocco (Tangier-Tetouan region).

INTRODUCTION: Among the most useful biological examinations in common medical practice, blood count is the most prescribed. The reference intervals of the hematological parameters of this examination are of major importance for clinical orientations and therapeutic decisions. In Morocco, the reference values used by the laboratories of medical biology and used by doctors are ones collected from Caucasian and European individuals. These values could be different in the Moroccan population. Besides, reference intervals of the blood count specific to the various Moroccan regions are missing. We decided to determine the reference intervals from a population of healthy adults of the Tangier-Tetouan region by following the procedures recommended by the IFCC-CLSI guidelines in 2008 and comparing them to those of the literature. METHODS: Blood samples were taken from 15840 adult volunteers (8402 men from 18 to 55 years old and 7438 women from 18 to 50 years old) from the regional transfusion center of Tangier and Tetouan during a period between November 2014 and May 2016. The complete blood count was measured by the Sysmex KX21N® analyzer. For each sample a systematic blood smear was done to determine the leukocyte differential. The data analysis was made by the software SPSS 20.0 by using percentiles 2.5th and 97.5th. RESULTS: A significant difference between both sexes was noted (p<0,001) for all the hematological parameters (red blood cells, hematocrit, hemoglobin, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, leukocytes, neutrophils, basophils, eosinophils, monocytes, platelets and mean platelet volume) except for the numeration of lymphocytes (p = 0.552). The values of this study were compared with those reported in Arabic, Caucasian and African populations. Said comparisons showed the existence of significant differences. CONCLUSION: This study tries to accentuate the necessity of proceeding with the establishment of reference intervals specific to the blood count of the Moroccan population to avoid errors of diagnosis, allow clinicians to interpret with greater specificity the hematological examinations and to improve the quality of medical care distributed to patients.

Pineal melatonin is a circadian time-giver for leptin rhythm in Syrian hamsters.

Nocturnal secretion of melatonin from the pineal gland may affect central and peripheral timing, in addition to its well-known involvement in the control of seasonal physiology. The Syrian hamster is a photoperiodic species, which displays gonadal atrophy and increased adiposity when adapted to short (winter-like) photoperiods. Here we investigated whether pineal melatonin secreted at night can impact daily rhythmicity of metabolic hormones and glucose in that seasonal species. For that purpose, daily variations of plasma leptin, cortisol, insulin and glucose were analyzed in pinealectomized hamsters, as compared to sham-operated controls kept under very long (16 h light/08 h dark) or short photoperiods (08 h light/16 h dark). Daily rhythms of leptin under both long and short photoperiods were blunted by pinealectomy. Furthermore, the phase of cortisol rhythm under a short photoperiod was advanced by 5.6 h after pinealectomy. Neither plasma insulin, nor blood glucose displays robust daily rhythmicity, even in sham-operated hamsters. Pinealectomy, however, totally reversed the decreased levels of insulin under short days and the photoperiodic variations in mean levels of blood glucose (i.e., reduction and increase in long and short days, respectively). Together, these findings in Syrian hamsters show that circulating melatonin at night drives the daily rhythmicity of plasma leptin, participates in the phase control of cortisol rhythm and modulates glucose homeostasis according to photoperiod-dependent metabolic state.

The circadian gene Clock oscillates in the suprachiasmatic nuclei of the diurnal rodent Barbary striped grass mouse, Lemniscomys barbarus: a general feature of diurnality?

A major challenge in the field of circadian rhythms is to understand the neural mechanisms controlling the oppositely phased temporal organization of physiology and behaviour between night- and day-active animals. Most identified components of the master clock in the suprachiasmatic nuclei (SCN), called circadian genes, display similar oscillations according to the time of day, independent of the temporal niche. This has led to the predominant view that the switch between night- and day-active animals occurs downstream of the master clock, likely also involving differential feedback of behavioral cues onto the SCN. The Barbary striped grass mouse, Lemniscomys barbarus is known as a day-active Muridae. Here we show that this rodent, when housed in constant darkness, displays a temporal rhythmicity of metabolism matching its diurnal behaviour (i.e., high levels of plasma leptin and hepatic glycogen during subjective midday and dusk, respectively). Regarding clockwork in their SCN, these mice show peaks in the mRNA profiles of the circadian gene Period1 (Per1) and the clock-controlled gene Vasopressin (Avp), which occur during the middle and late subjective day, respectively, in accordance with many observations in both diurnal and nocturnal species. Strikingly, expression of the circadian gene Clock in the SCN of the Barbary striped grass mouse was not constitutive as in nocturnal rodents, but it was rhythmic. As this is also the case for the other diurnal species investigated in the literature (sheep, marmoset, and quail), a hypothesis is that the transcriptional control of Clock within the SCN participates in the mechanisms underlying diurnality and nocturnality.

Daily behavioral rhythmicity and organization of the suprachiasmatic nuclei in the diurnal rodent, Lemniscomys barbarus.

Wheel-running activity was recorded in Lemniscomys barbarus exposed to different lighting conditions. This rodent shows rhythmic locomotor activity under natural twilight-light/dark (LD) as well as squared-LD cycles. A mean of 77% of the activity occurred during the light phase. Under different controlled photoperiods, the quantity of daily locomotor activity was relatively stable except for a lower level in the shortest photoperiod tested (LD 06:18). The duration of the active phase tended to increase with the duration of the light phase, especially in the longer photoperiods. Whatever the lighting conditions, Lemniscomys barbarus started running before lights-on and stopped after lights-off. The phase angle of activity offset relative to lights-off was stable in each squared-photoperiod, whereas the phase angle of activity onset relative to lights-on was significantly the highest under the shortest photoperiods. Recording of activity under constant lighting conditions showed that the daily rhythm of locomotor activity is fundamentally circadian. The endogenous period was slightly<24 h (mean=23.8 h) in permanent darkness and>24 h (mean=24.5 h) in continuous light. Re-entrainment of the locomotor activity rhythm after a 6 h phase advance or delay requires only four days on average. Moreover, the phase-responses curve to a 30 min light pulse (200 lux) in Lemniscomys barbarus kept in constant dark reveals large phase shifts according to circadian times (CT). With CT0 being defined as the onset of daily activity, maximum phase delay and advance shifts were observed at CT11 (Delta Psi=-5.7 h+/-2.3 h) and CT21 (Delta Psi =4.9+/-1.2 h), respectively. Interestingly, the phase-response curve to light did not show any dead zone. Immunohistochemical staining of the suprachiasmatic nuclei indicates that arginine vasopressin-immunoreactive cell bodies and fibers delimited a dorsal subregion that extends laterally and medially. The ventral subregion is rich in vasoactive intestinal peptide-immunoreactive neurones overlapping a smaller area containing gastrin-releasing peptide-expressing cells and receives numerous fibers labeled with neuropeptide Y antibody. The results of this study clearly demonstrate that Lemniscomys barbarus is a diurnal species highly sensitive to the shifting effects of light. Overall, this rodent can be considered a new and interesting model for circadian rhythm neurobiology.

Photoreceptor organisation and phenotypic characterization in retinas of two diurnal rodent species: potential use as experimental animal models for human vision research.

To characterize rod and cone distribution and composition in two diurnal mouse-like rodents, retinas from adult Arvicanthis ansorgei and Lemniscomys barbarus were processed for immunohistochemistry using multiple rod- and cone-specific antibodies. Antibodies tested included rhodopsin, cone opsins, pan-arrestin and cone arrestin, recoverin, and cGMP dependent ion channel. In both species, retinas were composed of approximately 33% cones, and most antibodies gave similar staining patterns. Data show these two diurnal rodents possess large numbers of cones, organised in a strict anatomical array. This suggests that diurnal rodents in general possess elevated cone numbers and could constitute valuable models for investigating cone pathophysiology.