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Biomedical subjects

M J Soares

Publications and source records attributed to M J Soares.

At least 37 records · Page 2Linked to original sources

The effect of leprosy-induced deformity on the nutritional status of index cases and their household members in rural South India: a socio-economic perspective.

OBJECTIVES: To determine whether the socioeconomic and nutritional status of cured leprosy patients with residual deformity, and their household members, was lower than that of cured leprosy patients without deformity. DESIGN: Cross-sectional study. SUBJECTS: One hundred and fifty-five index cases with deformity, 100 without deformity. Also 616 household members comprising 48% of the total members enumerated. MEASUREMENTS: Nutritional status was evaluated using anthropometry. Disease characteristics, socio-economic parameters and household information were recorded using a questionnaire. RESULTS: Index cases with deformity had lower community acceptance (P<0.001), and employment (P<0.001) than those cases without deformity. Households of index cases with deformity had a lower income (P<0.01) and a lower expenditure on food (P<0.05). The presence of deformity (odds ratio (OR): 2.1-3.2, P<0.01), unemployment (OR: 2.3-4.3, P<0.01) and female gender (OR: 2.4, P<0. 01) significantly increased the risk of index cases being undernourished, as judged by body mass index (BMI) alone, or BMI and mid-upper arm circumference. A low BMI (<18.5) in the index case significantly increased the odds of other adults (OR 2.2), adolescents (OR 2.9-3.8) and children (OR 2.2) in the household being undernourished. CONCLUSIONS: Cured leprosy index cases with physical deformity are more undernourished than index cases without deformity. This is associated with a reduced expenditure on food, possibly brought on by increased unemployment, and a loss of income. Undernutrition in the index case increases the risk of undernutrition in other members of the family. European Journal of Clinical Nutrition (2000) 54, 643-649.

Adolescent↗

Indirect estimates of body composition are useful for groups but unreliable in individuals.

OBJECTIVE: To assess the usefulness of the body mass index (BMI) in identifying individuals classified as overweight or obese based on estimates of body fat percentage (BF%) obtained by the deuterium dilution (BF%DD) method. In addition, to assess the accuracy of bioelectrical impedance analysis (BIA) and skinfold thickness (SFT) measurements in the estimation of body composition of Australians at the individual and group level. DESIGN: Cross-sectional study. SUBJECTS: One hundred and seventeen healthy Australian volunteers of European descent, comprising of 51 males and 66 females, ranging in age from 19 to 77 y. MEASUREMENTS: BMI was calculated from body weight and height. Fat-free mass (FFM) was estimated from measures of total body water (TBW) using deuterium dilution (FFM(DD)), SFT using the equations of Durnin and Womersley (Br J Nutr 1974; 32: 77-97) (FFM(SFT)), and BIA using the equations of Lukaski et al (J Appl Physiol 1986; 60: 1327-1332) (FFM(Lu)), Segal et al (Am J Clin Nutr 1988; 47: 7-14) (FFM(Se)) and Heitmann (Eur J Clin Nutr 1990; 44: 831-837) (FFM(He)). Estimates of fat mass (FM) were calculated as the difference between body weight and FFM, while BF% was calculated by expressing FM as a percentage of body weight. RESULTS: BMI had poor sensitivity and positive predictive value in identifying individuals as being overweight/obese as classified by BF%DD. Furthermore, estimates of FFM (and hence FM) from BIA or SFT could not be used interchangeably with DD, without the risk of considerable error at the individual level. At the group level errors were relatively smaller, though statistically significant. While FFM(SFT) could be corrected by the addition of the bias (1.2 kg in males and 0.8 kg in females), no simple correction was possible with BIA estimates of FFM for any of the equations used. However, an accurate prediction of FFM(DD) was possible from the combination of FFM(He), biceps SFT and mid-arm circumference in both males and females. The bias of this prediction was small (<0.15 kg), statistically non-significant in both sexes, and unrelated to the mean FFM obtained by the two methods. The revision of Heitmann's estimate of FFM using anthropometric variables described in this study had the best sensitivity (79%), specificity (96%) and positive predictive value (92%) in identifying overweight/obese individuals in comparison to the other equations tested. CONCLUSION: BMI was a poor surrogate for body fatness in both males and females. The currently recommended equations for the prediction of body composition from SFT and BIA provided inaccurate estimates of FFM both at the individual and group level as compared to estimates from DD. However, Heitmann's equations, when combined with measures of the biceps SFT and mid-arm circumference, provided better estimates of FFM both at the individual and group level.

Adipose Tissue↗

Plasma leptin concentrations, basal metabolic rates and respiratory quotients in young and older adults.

OBJECTIVE: To-examine the interrelationship of circulating leptin concentrations, basal metabolic rates (BMR) and respiratory quotients (RQ) in young and older adults. DESIGN: Cross sectional study. SUBJECTS: Seventy-six Australian men and women, 48 young (< 35 y) and 28 older ( > or = 50 y). MEASUREMENTS: Fasting plasma leptin concentrations by RIA, BMR and RQ by indirect calorimetry, percentage body fat (BF%), fat mass (FM) and fat-free mass (FFM) from total body water (TBW) based on deuterium dilution, waist and hip circumferences from anthropometry. RESULTS: Older subjects had significantly higher BF%, FM and waist-to-hip ratio (WHR), but significantly lower FFM and absolute BMR as compared to younger subjects. Absolute leptin concentrations were 60% higher in older subjects but did not achieve statistical significance. There was, however, a significant gender x age group interaction in leptin concentrations. This was reflected in a significant inverse relationship between age group and leptin in women when data was controlled for waist circumference (r = -0.38, P = 0.028), or FM (r = -0.36, P = 0.042). A similar relationship was not observed in men on controlling for BF% or FM. Log transformed plasma leptin was best explained by a model that included BF%, gender, age-group, gender x age-group and WHR r = 0.75, adjusted r2 = 0.56, standard error of estimate (SEE) = 0.73 ng/ml). BMR was best explained by FFM, FM and age group r = 0.94, adjusted r2 = 0.87; SEE = 429 kJ/day). On controlling for BF%, WHR and FFM, leptin was negatively related to RQ only in older men (r = -0.67, P = 0.033). There was no relationship of leptin to BMR in the groups studied. CONCLUSION: The study demonstrates an age-related modification of the gender bias in leptin, and a gender-specific inverse relationship between leptin and RQ in older people. The decline in leptin and the lack of a relationship between RQ and leptin in older women may indicate an increased risk of weight gain relative to older men.

Adipose Tissue↗

Fatty acid transport regulatory proteins in the developing rat placenta and in trophoblast cell culture models.

The placenta forms a selective barrier that is able to transport nutrients that are of critical use to the fetus. Delivery of essential fatty acids to the fetus is dependent upon transplacental transport and provides the backbone for the biosynthesis of biological membranes, myelin and various signalling molecules. The primary objective of this research was to elucidate the expression patterns of genes that regulate fatty acid transport across the placenta. Several fatty acid transport regulatory genes have been identified in the rat including; cytoplasmic heart fatty acid binding protein (hFABP), plasma membrane fatty acid binding protein (FABPpm), fatty acid translocase (FAT) and fatty acid transport protein (FATP). In this study, we have elucidated temporal and spatial expression patterns for these genes in the rat placenta and in cell culture models of the rat placenta by Northern blot, RT-PCR, Western blot and/or by in situ hybridization analyses. Expression of hFABP was specific to the labyrinth zone, the main barrier and site of transplacental transport in the rat placenta. In addition, the levels of hFABP expression increased with gestational age, suggesting a growing requirement for fatty acid transport with advancing stages of pregnancy. FABPpm, FAT and FATP are expressed in both the junctional and labyrinth zones of the rat placenta. FAT was predominantly localized to the labyrinth zone by in situ hybridization analysis. The placental cell expression patterns of the genes involved in fatty acid transport were supported by our observations of HRP-1 (labyrinth zone) and Rcho-1 (junctional zone) trophoblast cell culture models. Given their cell surface location, we predict that FABPpm, FAT and FATP potentially participate in placental fatty acid uptake. The predominant expression of hFABP and FAT in the labyrinth zone of the chorioallantoic placenta implicates hFABP and FAT in the transplacental movement of fatty acids from maternal to fetal compartments.

Animals↗

Sensilla on the terminalia of Lutzomyia spp. (Diptera: Psychodidae) sand flies.

Sensilla on the male terminalia of four different species of sand flies (Diptera: Psychodidae) are described. Two species belonged to the same species complex: Lutzomyia longipalpis (Lutz & Neiva) and L. cruzi (Mangabeira). Other Brazilian species complex was represented by L. lenti (Mangabeira) and L. carmelinoi (Ryan, Fraiha, Lainson & Shaw). Approximately 5-15 sensilla were found on the initial part of the styles of all species. Structurally, the sensory organs were similar to small coeloconica sensilla. Their number varied both individually and interspecifically. These sensilla may play an important role in the copulation behavior of these species of sand flies.

Animals↗

Differentiation of Trypanosoma cruzi epimastigotes: metacyclogenesis and adhesion to substrate are triggered by nutritional stress.

Differentiation of Trypanosoma cruzi epimastigotes to metacyclic trypomastigotes occurs in the insect rectum, after adhesion of the epimastigotes to the intestinal wall. We investigated the effect of the nutritional stress on the metacyclogenesis process in vitro by incubating epimastigotes in the chemically defined TAU3AAG medium supplemented with different nutrients. Addition of fetal bovine serum induced epimastigote growth but inhibited metacyclogenesis. In this medium, few parasites attached to the substrate. Ultrastructural analysis demonstrated reservosomes at the posterior end of the epimastigotes. Incubation of the cells in TAU3AAG medium containing gold-labeled transferrin resulted in high endocytosis of the marker by both adhered and free-swimming epimastigotes. No intracellular gold particles could be detected in trypomastigotes. Addition of transferrin gold complexes to adhered epimastigotes cultivated for 4 days in TAU3AAG medium resulted in decrease of both metacyclogenesis and adhesion to the substrate, as compared with parasites maintained in transferrin-free medium. Adhesion to the substrate is triggered by nutritional stress, and proteins accumulated in reservosomes are used as energy source during the differentiation. A close relationship exists among nutritional stress, endocytosis of nutrients, adhesion to the substrate, and cell differentiation in T. cruzi epimastigotes.

Animals↗

Identification of two new nonclassical members of the rat prolactin family.

The prolactin (PRL) family is comprised of a group of hormones/cytokines that are expressed in the anterior pituitary, uterus, and placenta. These proteins participate in the control of maternal and fetal adaptations to pregnancy. In this report, we have identified two new nonclassical members of the rat PRL family through a search of the National Center for Biotechnology Information dbEST database. The cDNAs were sequenced and their corresponding mRNAs characterized. Overall, the rat cDNAs showed considerable structural similarities with mouse proliferin-related protein (PLF-RP) and prolactin-like protein-F (PLP-F), consistent with their classification as rat homologs for PLF-RP and PLP-F. The expression of both cytokines/hormones was restricted to the placenta. The intraplacental sites of PLF-RP and PLP-F synthesis differed in the rat and the mouse. In the mouse, PLF-RP was expressed in the trophoblast giant cell layer of the midgestation chorioallantoic and choriovitelline placentas and, during later gestation, in the trophoblast giant cell and spongiotrophoblast layers within the junctional zone of the mouse chorioallantoic placenta. In contrast, in the rat, PLF-RP was first expressed in the primordium of the chorioallantoic placenta (ectoplacental cone region) and, later, exclusively within the labyrinth zone of the chorioallantoic placenta. In the mouse, PLP-F is an exclusive product of the spongiotrophoblast layer, whereas in the rat, trophoblast giant cells were found to be the major source of PLP-F, with a lesser contribution from spongiotrophoblast cells late in gestation. In summary, we have established the presence of PLF-RP and PLP-F in the rat.

Amino Acid Sequence↗

Induction of erythroid proliferation and differentiation by a trophoblast-specific cytokine involves activation of the JAK/STAT pathway.

Pregnancy is characterized by increased erythropoiesis within maternal and fetal compartments. The placenta has been shown to produce factors that stimulate erythropoiesis but convincing evidence for placental production of erythropoietin (EPO) is still lacking. Prolactin-like protein E (PLP-E) was recently found to stimulate expression of the adult beta major globin gene in mouse erythroleukemia cells. Here we demonstrate that PLP-E transiently expressed in COS-7 cells stimulates proliferation and erythroid differentiation of murine and human erythroid progenitor cell lines. Electrophoretic mobility shift assays were used to show the activation of STAT5 by PLP-E in the human erythroid cell line TF1. Furthermore, we compared the effects of PLP-E on murine myeloid FDCP1 cells which do not express EPO receptors (EPORs) with effects on cells genetically engineered to express functional EPORs. We provide evidence that PLP-E-dependent proliferation and STAT5 activation is independent of the expression of the EPOR. Taken together, these data suggest that PLP-E acts on specific receptors of erythroid-committed murine and human cells by the activation of intracellular signaling pathways promoting cell growth and differentiation.

Animals↗

Placental lactogen-I gene activation in differentiating trophoblast cells: extrinsic and intrinsic regulation involving mitogen-activated protein kinase signaling pathways.

Trophoblast giant cells are one of the primary endocrine cell types of the rodent placenta. Placental lactogen-I (PL-I) is the initial prolactin (PRL) family member expressed as trophoblast giant cells differentiate. In this report, we use the Rcho-1 trophoblast cell line as a model for studying the regulation of PL-I gene expression during trophoblast giant cell differentiation. Evidence is provided for trophoblast cell expression of epidermal growth factor receptor (EGFR), ErbB2, fibroblast growth factor receptor 1 (FGFR1), transforming growth factor-alpha, and heparin-binding EGF. EGF and FGF-2 stimulated PL-I mRNA and protein accumulation and PL-I promoter activity in a concentration-dependent manner. These latter growth factor actions on PL-I promoter activities were specifically inhibited by cotransfection with dominant negative constructs for EGFR and FGFRs respectively. Utilization of the mitogen-activated protein kinase (MAPK) pathway by EGF and FGF-2 in trophoblast cells was demonstrated by growth factor stimulation of a Gal4 DNA binding/Elk1 transactivational domain fusion construct, and more specifically by activation of extracellular signal regulated kinase and p38 MAPK. PL-I gene activation was also sensitive to disruption of MAPK and activation protein-1 (AP-1) signaling pathways. In conclusion, autocrine/paracrine pathways involving EGFR and FGFR1, MAPK and AP-1 are shown to participate in the regulation of the PL-I gene in differentiating trophoblast cells.

Analysis of Variance↗

Three novel paralogs of the rodent prolactin gene family.

The prolactin (PRL) family consists of a collection of genes expressed in the uterus, placenta and anterior pituitary. These cytokines/hormones participate in the control of maternal-fetal adaptations to pregnancy. In this report, we establish the presence of three new members of the PRL family. Novel expressed sequence tags (ESTs) with homology to PRL were isolated from embryonic and placental cDNA libraries. The cDNAs were sequenced and compared with those of other members of the PRL family. The three new cDNAs were assigned to the PRL family on the basis of sequence similarities and were referred to as PRL-like protein-J (PLP-J), PRL-like protein-K (PLP-K) and PRL-like protein-M (PLP-M). Both rat and mouse PLP-J cDNAs were identified. Rat PLP-J cDNA encodes for a predicted 211 amino acid protein containing a 29 amino acid signal peptide and two putative N-linked glycosylation sites, whereas the mouse PLP-J cDNA encodes for a 212 amino acid protein containing a 29 amino acid signal peptide with a single N-linked glycosylation site. Rat and mouse PLP-J proteins share approximately 79% and 70% nucleotide and amino acid sequence identity, respectively. A full-length rat PLP-K cDNA and a partial tentative mouse PLP-K cDNA were identified. The rat PLP-K cDNA encodes for a predicted 228 amino acid protein containing a 31 amino acid signal peptide and one putative N-linked glycosylation site; the mouse PLP-M cDNA encodes for a predicted 228 amino acid protein containing a 28 amino acid signal peptide and one putative N-linked glycosylation site. Genes for PLP-J, PLP-K and PLP-M are situated at the Prl family locus on mouse chromosome 13. PLP-J was exclusively expressed in decidual tissue from both the mouse and rat. PLP-K was expressed in trophoblast cells of the chorioallantoic placenta and showed an apparent species difference. In the mouse, virtually all trophoblast lineages expressed PLP-K, whereas in the rat, PLP-K expression was restricted to the labyrinthine trophoblast cells. Mouse PLP-M expression was restricted to the junctional zone of the chorioallantoic placenta. In summary, we have identified three new members of the rodent PRL gene family that are expressed in uterine and placental structures. Future experimentation is needed to determine the specific roles of each of these ligands in the biology of pregnancy.

Amino Acid Sequence↗

Eosinophils are cellular targets of the novel uteroplacental heparin-binding cytokine decidual/trophoblast prolactin-related protein.

The uterus and placenta of the mouse and rat produce a member of the prolactin (PRL) family referred to as decidual/trophoblast PRL-related protein (d/tPRP). This cytokine/hormone has been hypothesized to regulate decidual cell activities needed for the establishment and maintenance of gestation. An alkaline phosphatase (AP)-tagging strategy was used to identify d/tPRP target cells. AP-d/tPRP bound to virtually all cells and tissues to which it was exposed, consistent with our earlier evidence that d/tPRP binds to heparin-containing molecules. Moreover, we found that co-incubation with heparin or pretreatment with heparitinase greatly decreased the binding of AP-d/tPRP to tissue sections. In addition, we observed that the AP-d/tPRP probe bound to the surface of Chinese hamster ovary (CHO) cells but not to heparan sulfate-deficient CHO-pgsD-677 cells. Potential unique non-heparin d/tPRP binding sites within mouse and rat uteroplacental tissues were identified by consecutively incubating sections with AP-d/tPRP followed by heparin. This strategy led to the identification of d/tPRP target cells associated with the uterus and the labyrinth zone of the chorioallantoic placenta. Within the uterus, d/tPRP specifically bound to eosinophils. d/tPRP-binding and eosinophil peroxidase activity were co-localized and showed similar patterns of distribution during the estrous cycle, pregnancy, and following hormonal manipulation. d/tPRP interactions with eosinophils were further demonstrated in the lung and intestine, with eosinophils isolated from the peritoneum, and in mice with generalized tissue eosinophilia. Collectively, these findings suggest that intercellular d/tPRP targeting is mediated through associations with heparin-containing molecules which help direct d/tPRP to specific interactions with eosinophils within the uterus and with the labyrinthine compartment of the chorioallantoic placenta.

Animals↗

Differentiation-dependent expression of gelatinase B/matrix metalloproteinase-9 in trophoblast cells.

The purpose of this study was to evaluate the Rcho-1 trophoblast culture system as a model for studying trophoblast invasion and to examine stage-specific expression of enzyme(s) potentially participating in rat trophoblast giant cell invasive behavior. The invasive behavior of the differentiating Rcho-1 trophoblast cells was demonstrated using Matrigel invasion chambers. Gelatin zymography and Western blot analysis of conditioned medium from differentiating Rcho-1 trophoblast cell cultures and rat ectoplacental cone outgrowths revealed a differentiation-dependent increase in gelatinase B/matrix metalloproteinase (MMP-9). Nothern blot and reverse transcriptase polymerase chain reaction (RT-PCR) analyses of Rcho-1 trophoblast or ectoplacental cone cells also showed increasing expression of MMP-9 accompanying cell differentiation. Rcho-1 trophoblast cells stably transfected with MMP-9 promoter/luciferase reporter constructs exhibited a differentiation-dependent increase in MMP-9 promoter activation. In conclusion, trophoblast giant cell differentiation is characterized by transcriptional activation of the MMP-9 gene and appearance of the invasive phenotype.

Animals↗

Immunocytochemical mapping of the hemoglobin biosynthesis site in amphibian erythroid cells.

During the past 25 years, several studies have attempted to determine the site of integration of the heme and the four globin chains in vertebrate erythroid cells that is important in the formation of the hemoglobin molecule. Mitochondrion-like organelles or hemosomes were pointed out as responsible for this task. We performed several experiments to investigate this hypothesis. The intracellular distribution of hemoglobin in amphibian erythroid cells was detected by post-embedding immuno-electron microscopy, using a polyclonal anti-human hemoglobin-proteinA-gold complex. Hemoglobin mapping showed an intense labeling in the cell cytoplasm, but none in cytoplasmic structures such as endoplasmic reticulum, mitochondria, mitochondrion-like organelles, Golgi complex, ribosomes or ferruginous inclusions. The mitochondrial fraction obtained according to the protocol described for some authors, showed by ultrastructural examination that this fraction has a heterogeneous content, also composed by microvesicles rich in cytoplasmic hemoglobin, an artifact generated by mechanical action during cell fractionation. Thus, when this fraction is lysed and its content submitted to electrophoresis, hemoglobin bands would be found inevitably, causing false-positive results, erroneously attributed to hemoglobin content of mitochondrion-like organelles. Our data do not confirm the hypothesis that the final hemoglobin biosynthesis occurs inside mitochondrion-like organelles. They suggest that the hemoglobin molecule be assembled in the erythrocyte cytoplasm outside of mitochondria or hemosomes.

Animals↗

Using consensus OPCRIT diagnoses. An efficient procedure for best-estimate lifetime diagnoses.

BACKGROUND: The Operational Criteria Checklist (OPCRIT) generates diagnoses according to 12 operational diagnostic systems (e.g. DSM-III, DSM-III-R, Research Diagnostic Criteria, ICD-10). AIMS: To examine the agreement between diagnoses generated by the OPCRIT, as completed by the interviewer, with a best-estimate lifetime procedure using the OPCRIT. METHOD: Subjects came from large multi-generational bipolar or schizophrenia pedigrees (n = 100), and from a sample of unrelated subjects with schizophrenia (n = 40). We analysed the diagnostic agreement between OPCRIT diagnoses generated by the interviewer and our best-estimate OPCRIT diagnoses, according to DSM-III-R and ICD-10, using Cohen kappa statistics. RESULTS: Excellent agreement was found between interviewer OPCRIT diagnoses and OPCRIT diagnoses made by the best-estimate lifetime consensus procedure for DSM-III-R (kappa = 0.83) and ICD-10 (kappa = 0.81). CONCLUSIONS: Results suggest that this procedure for diagnostic assessment is an efficient alternative to classic best-estimate diagnosis procedures.

Algorithms↗

Distinct regulatory regions from the prolactin-like protein C variant promoter direct trophoblast giant cell versus spongiotrophoblast cell-specific expression.

PRL-like protein C variant (PLP-Cv) is a newly identified member of the PRL family. PLP-Cv is specifically expressed in the chorioallantoic placenta by two distinct cell populations: trophoblast giant cells and spongiotrophoblast cells. To gain some insight regarding the control of PLP-Cv gene expression and the regulatory factors controlling trophoblast giant cell and spongiotrophoblast cell lineages, we have initiated a structural and functional analysis of the PLP-Cv promoter. The activities of a series of PLP-Cv promoter constructs, ranging in size from 4.5 kb to 50 bp, ligated to a luciferase reporter have been assessed in the Rcho-1 trophoblast cell line (restricted to trophoblast giant cell differentiation) and in a primary spongiotrophoblast cell culture system after transient transfection. PLP-Cv promoter constructs containing 4.5 kb to 149 bp of 5'-flanking DNA possessed full activity in the trophoblast giant cell model. A region located between -149 and -124 bp upstream of the PLP-Cv transcription start site was found to be essential for activation of the PLP-Cv promoter. Spongiotrophoblast cells required additional PLP-Cv 5'-flanking DNA for full activity. A region located between -2518 and -2242 bp upstream of the PLP-Cv transcription start site significantly enhanced PLP-Cv promoter in spongiotrophoblast cells. In conclusion, mechanisms underlying the activation of the PLP-Cv promoter are different in trophoblast giant cells vs. spongiotrophoblast cells.

Animals↗

Uterine natural killer cells are targets for a trophoblast cell-specific cytokine, prolactin-like protein A.

PRL-like protein A (PLP-A) is a member of the PRL family expressed in trophoblast cells coincident with establishment of the chorioallantoic placenta. The purpose of this investigation was to identify targets for PLP-A. Using an alkaline phosphatase-tagging strategy, we show that PLP-A specifically interacts with a population of natural killer (NK) lymphocytes within the mesometrial compartment of decidua from pregnant and pseudopregnant rats. These observations are supported by the codistribution of PLP-A targets with cells expressing the rat NK cell surface marker, gp42, the absence of PLP-A binding in conceptuses from NK cell-deficient tg epsilon26 mice, and the specific interaction of PLP-A with a rat NK cell line, RNK-16. We have further demonstrated that PLP-A effectively suppresses RNK-16 cell cytolytic activities. Our results provide evidence for a new paradigm of embryonic-maternal communication involving a PLP-A signaling pathway between trophoblast cells and uterine NK lymphocytes.

Alkaline Phosphatase↗