Molecular insights and antibody response to Dr20/22 in dogs naturally infected with Dirofilaria repens.

Subcutaneous dirofilariasis, caused by the parasitic nematode Dirofilaria repens, is a growing concern in Europe, affecting both dogs and humans. This study focused on D. repens Dr20/22, a protein encoded by an alt (abundant larval transcript) gene family. While well-documented in L3 larvae of other filariae species, this gene family had not been explored in dirofilariasis. The research involved cloning Dr20/22 cDNA, molecular characterization, and evaluating its potential application in the diagnosis of dirofilariasis. Although Real-Time analysis revealed mRNA expression in both adult worms and microfilariae, the native protein remained undetected in lysates from both developmental stages. This suggests the protein's specificity for L3 larvae and may be related to a process called SLTS (spliced leader trans-splicing), contributing to stage-specific gene expression. The specificity of the antigen for invasive larvae positions it as a promising early marker for dirofilariasis. However, ELISA tests using sera from infected and uninfected dogs indicated limited diagnostic utility. While further research is required, our findings contribute to a deeper understanding of the molecular and immunological aspects of host-parasite interactions and could offer insights into the parasite's strategies for evading the immune system.

Beyond Tradition: Exploring Cutting-Edge Approaches for Accurate Diagnosis of Human Filariasis.

Filariasis is recognised as a global public health threat, particularly in tropical and subtropical regions. It is caused by infection with a nematode parasite of the superfamily Filarioidea, including Wuchereria bancrofti, Brugia malayi, Onchocerca volvulus, and Onchocerca lupi. Three main types of filariasis have been classified: lymphatic filariasis, subcutaneous filariasis, and serous cavity filariasis. The symptoms exhibited by individuals afflicted with filariasis are diverse and contingent upon several variables, including the species of parasite, the host's health and immune response, and the stage of infection. While many classical parasitological techniques are considered indispensable tools for the diagnosis of parasitic infections in humans, alternative methods are being sought due to their limitations. Novel tests based on host-parasite interactions offer a rapid, simple, sensitive, and specific diagnostic tool in comparison to traditional parasitological methods. This article presents methods developed in the 21st century for the diagnosis of filariasis caused by invasion from W. bancrofti, B. malayi, O. volvulus, and O. lupi, as well as techniques that are currently in use. The development of modern diagnostic methods based on molecular biology constitutes a significant advancement in the fight against filariasis.

Canine antibody response against Dirofilaria repens in natural occult and microfilaremic infections.

Dirofilaria repens is the causative agent of the zoonotic canine skin condition called subcutaneous dirofilariosis. Despite being endemic throughout much of Europe, little is known about host humoral responses to D. repens. To address this, we analyzed serum immunoglobulin isotypes recognizing D. repens somatic antigens (DrSA) in naturally infected dogs. Titers of anti-DrSA IgG and IgE, but not IgM were significantly higher in dogs infected with D. repens compared to those that were negative. Moreover, microfilaremic infections were associated with higher levels of IgG1 than IgG2, while occult infections occurred with significantly higher levels of IgG2 than IgG1. Finally, the measurement of anti-DrSA IgG antibodies allowed the detection of occult subcutaneous dirofilariosis in dogs.

Selection of new diagnostic markers for Dirofilaria repens infections with the use of phage display technology.

Dirofilaria repens is a parasitic nematode causing vector-borne disease (dirofilariasis), considered an emerging problem in veterinary and human medicine. Although main hosts are carnivores, particularly dogs, D. repens shows high zoonotic potential. The disease spreads uncontrollably, affecting new areas. Since there is no vaccine against dirofilariasis, the only way to limit disease transmission is an early diagnosis. Currently, diagnosis depends on the detection of microfilariae in the host bloodstream using modified Knott's test or multiplex PCR. However, the efficacy of tests relying on microfilariae detection is limited by microfilariae periodic occurrence. Therefore, a new reliable diagnostic test is required. Our study aimed to select new diagnostic markers for dirofilariasis with potential application in diagnostics. We focused on single epitopes to ensure high specificity of diagnosis and avoid cross-reactivity with the other parasite infections common in dogs. Using phage display technology and 12-mer peptides library, we selected epitopes highly reactive with IgG from sera of infected dogs. Additionally, our study presents the possibility of detecting D. repens specific cell-free DNA in dogs with no microfilaria but high IgG and IgM antibody levels against parasite somatic antigen.

The Immunological Role of Vascular and Lymphatic Endothelial Cells in Filarial Infections.

The embryonic stage of filarial nematodes, or microfilariae (Mf), shows daily and seasonal periodicity that requires their migration through blood vessels into the lungs, where they are sequestered when not circulating in the peripheral blood. Therefore, Mf and the host endothelium are likely in a permanent state of hide and seek. Interestingly, filarial nematodes co-cultured in media with a murine endothelial cell line survive eight times longer than those cultured in media alone. This suggests that the endothelium is an important element of the immune response in filarial nematodes, perversely promoting their survival in the host. In this review, we will focus on potential pathways involved in the relationship between filarial nematodes and the host endothelium, including the role of endothelial ICAM/VCAM/PECAM adhesion molecules, surface markers involved in the passage of Mf through host tissue, anti-thrombolic effects caused by the presence of filarial nematodes (including plasmins), endothelial cell proliferation (VEGF), and other aspects of the immune activation of the endothelium. The aim of this review is to merge the knowledge about the cross-talk between Mf of different filarial nematode species and endothelial cells (EC), thus allowing a better understanding of the mechanism of these parasitic infections.

Nematode Orthologs of Macrophage Migration Inhibitory Factor (MIF) as Modulators of the Host Immune Response and Potential Therapeutic Targets.

One of the adaptations of nematodes, which allows long-term survival in the host, is the production of proteins with immunomodulatory properties. The parasites secrete numerous homologs of human immune mediators, such as macrophage migration inhibitory factor (MIF), which is a substantial regulator of the inflammatory immune response. Homologs of mammalian MIF have been recognized in many species of nematode parasites, but their role has not been fully understood. The application of molecular biology and genetic engineering methods, including the production of recombinant proteins, has enabled better characterization of their structure and properties. This review provides insight into the current state of knowledge on MIF homologs produced by nematodes, as well as their structure, enzymatic activity, tissue expression pattern, impact on the host immune system, and potential use in the treatment of parasitic, inflammatory, and autoimmune diseases.

Immunoproteomic Analysis of Dirofilaria repens Microfilariae and Adult Parasite Stages.

Dirofilariarepens is a parasitic nematode causing a vector-borne zoonotic infection (dirofilariosis), considered an emerging problem in human and veterinary medicine. Currently, diagnosis is based on the detection of the adult parasite and microfilariae in the host tissues. However, the efficacy of tests relying on microfilariae detection is limited by microfilariae periodic occurrence. Therefore, a new reliable and affordable serological diagnostic method is needed. Better characteristic of the parasite biology and its interaction with host immune system should help to achieve this goal. This study analyzes adult and microfilariae proteomes, and the use of one-dimensional electrophoresis (1-DE) and two-dimensional electrophoresis (2-DE) proteomics, immunoproteomics, and LC-MS/MS mass spectrometry allowed us to identify 316 potentially immunogenic proteins (75 belong to adult stage, 183 to microfilariae, and 58 are common for both). Classified by their ontology, the proteins showed important similarities and differences between both parasite stages. The most frequently identified proteins are structural, metabolic, and heat shock proteins. Additionally, real-time PCR analysis of some immunogenic targets revealed significant differences between microfilariae and adult life stages. We indicated molecules involved in parasite-host interactions and discussed their importance in parasite biology, which may help to reveal potential diagnostic antigens or select drug and vaccine targets.

Quantitative anatomy of the fused ossification center of the occipital squama in the human fetus.

CT-based quantitative analysis of any ossification center in the cranium has not previously been carried out due to the limited availability of human fetal material. Detailed morphometric data on the development of ossification centers in the human fetus may be useful in the early detection of congenital defects. Ossification disorders in the cranium are associated with either a delayed development of ossification centers or their mineralization. These aberrations may result in the formation of accessory skull bones that differ in shape and size, and the incidence of which may be misdiagnosed as, e.g., skull fractures. The study material comprised 37 human fetuses of both sexes (16♂, 21♀) aged 18-30 weeks. Using CT, digital image analysis software, 3D reconstruction and statistical methods, the linear, planar and spatial dimensions of the occipital squama ossification center were measured. The morphometric characteristics of the fused ossification center of the occipital squama show no right-left differences. In relation to gestational age, the ossification center of the occipital squama grows linearly in its right and left vertical diameters, logarithmically in its transverse diameters of both the interparietal and supraoccipital parts and projection surface area, and according to a quadratic function in its volume. The obtained numerical findings of the occipital squama ossification center may be considered age-specific references of relevance in both the estimation of gestational age and the diagnostic process of congenital defects.

Community Pharmacies in Poland-The Journey from a Deregulated to a Strictly Regulated Market.

Community pharmacies are the primary entities providing drugs to individual patients in Poland. The pharmacy market has been changing for many years due to significant changes in market regulations. These changes significantly affect the profitability of pharmacies, which may impact the quality of pharmacotherapy. The small number of pharmacies, which resulted from changes in the law in 2017, can influence the level of patient care. The article presents the community pharmacies market in Poland. Particular attention is paid to the legal regulations affecting community pharmacies and the impact of these regulations on the overall shape of the market. The Polish system's specificity, including the pharmacy market indicators, has been compared with data from other European Union countries.

Hematological and Biochemical Changes in Dogs Naturally Infected With Dirofilaria repens.

Subcutaneous dirofilariosis is a zoonotic disease emerging throughout Europe caused by the filarial nematode Dirofilaria repens. Despite its increasing prevalence, there is a large gap in knowledge of the impact of the parasite on the host. Currently classified as being non-pathogenic, recent evidence suggests that skin dirofilariosis is associated with dermatological conditions including concomitant pruritus, neoplastic processes, inflammation, and even blindness in dogs and humans. The aim of this study was to determine if natural canine D. repens infection leads to biological changes in the canine host. In a real-life veterinary clinic setting, animals are often presented to clinicians for unrelated issues, and D. repens is incidentally identified during inspection. As such, we compared hematological and biochemical parameters of 218 uninfected and 197 dogs naturally infected with D. repens. Interestingly, animals infected with D. repens had lower numbers of lymphocytes (p < 0.001), red blood cells (p < 0.001), and thrombocytes (p = 0.025), decreased hematocrit (p < 0.001), and increased alkaline phosphatase (p = 0.016) and creatinine activity (p = 0.023) compared to uninfected dogs. We further selected a subpopulation of 214 dogs having prima facie hematological and biochemical results within normal reference ranges to evaluate the effect of D. repens infections in seemingly healthy dogs. Among these patients, 93 dogs infected with D. repens had lower numbers of lymphocytes (p = 0.031), red blood cells (p = 0.025), and hematocrit (p = 0.002), higher glucose levels (p = 0.023), and border line elevated alkaline phosphatase levels (p = 0.054) compared to 121 uninfected animals. Despite being categorized as asymptomatic, we have observed hematological and biochemical changes associated with D. repens infections in dogs, and our data suggest that dirofilariosis may induce a state of chronic stress. These results link the presence of skin dirofilariosis to biological changes in the canine host, suggesting a mechanism for pathogenicity and shedding new light on the host-parasite relationship.

Case Studies of Severe Microfilaremia in Four Dogs Naturally Infected With Dirofilaria repens as the Primary Disease or a Disease Complicating Factor.

Subcutaneous dirofilariosis in dogs, caused by Dirofilaria repens, is an underdiagnosed disease, now recognized for its zoonotic potential, and growing distribution and prevalence across Europe and Asia. Our understanding of the pathogenicity in human and canine host remains unclear, but case reports suggest that microfilariae (Mf) as well as adult D. repens may directly cause internal organs damage or may be a factor complicating the course of other ailments. The purpose of the study was to report high Mf in dogs and to discuss potential relevance with co-morbidity. Our data from a modified Knott's test performed on 62 infected dogs indicate that the median Mf count in D. repens infections is 675 Mf/ml and we consider microfilaremia above 10,000 Mf/ml as high intensity. This collection of case reports discusses 4 cases of high intensity D. repens microfilaremia in companion dogs; one presenting pathology from a very high intensity of adult D. repens with post-treatment complications, and 3 dogs in which high microfilaremia was detected incidentally during the management of other primary illnesses. To our knowledge this report describes the highest D. repens microfilaremia ever detected in a dog, at 178,000 Mf/ml. The issue of high microfilaremic infections in dogs is poorly studied and there is growing need to identify the presentation and understand the mechanisms of associated pathogenesis in the host-parasite relationship.

Morphometric study of the primary ossification center of the frontal squama in the human fetus.

PURPOSES: Detailed morphometric data on the development of ossification centers in human fetuses is useful in the early detection of skeletal dysplasias associated with a delayed development of ossification centers and their mineralization. Quantitative analysis of primary ossification centers of cranial bones is sporadic due to limited availability of fetal material. MATERIAL AND METHODS: The size of the primary ossification center of the frontal squama in 37 human (16 males and 21 females) spontaneously aborted human fetuses aged 18-30 weeks was studied by means of CT, digital-image analysis and statistics. RESULTS: With neither sex nor laterality differences, the best-fit growth dynamics for the primary ossification center of the frontal squama was modelled by the following functions: y = 13.756 + 0.021 × (age)2 ± 0.024 for its vertical diameter, y = 0.956 + 0.956 × age ± 0.823 for its transverse diameter, y = 38.285 + 0.889 × (age)2 ± 0.034 for its projection surface area, and y = 90.020 + 1.375 × (age)2 ± 11.441 for its volume. CONCLUSIONS: Our findings for the primary ossification center of the frontal squama may be conducive in monitoring normal fetal growth and screening for inherited faults and anomalies of the skull in human fetuses.

Quantitative anatomy of the primary ossification center of the radial shaft in human fetuses.

PURPOSE: The medical literature still lacks studies on the size of the radial shaft primary ossification center, thus preventing us from potentially relevant data in diagnosing skeletal dysplasias, i.e., TAR syndrome, VATER syndrome, Holt-Oram syndrome, Fanconi anemia and Edwards syndrome, frequently characterized by disrupted or retarded fetal growth. MATERIALS AND METHODS: The size of the radial shaft primary ossification center in 47 (25 males and 22 females) spontaneously aborted human fetuses aged 17-30 weeks was studied by means of CT, digital image analysis and statistics. RESULTS: With neither sex nor laterality differences, the best-fit growth dynamics for the radial shaft primary ossification center was modeled by the following functions: y = - 10.988 + 1.565 × age ± 0.018 for its length, y = - 2.969 + 0.266 × age ± 0.01 for its proximal transverse diameter, y = - 0.702 + 0.109 × age ± 0.018 for its middle transverse diameter, y = - 2.358 + 0.203 × age ± 0.018 for its distal transverse diameter, y = -189.992 + 11.788 × (age)2 ± 0.018 for its projection surface area, and y = - 798.174 + 51.152 × age ± 0.018 for its volume. CONCLUSIONS: The morphometric characteristics of the radial shaft primary ossification center show neither sex nor bilateral differences. The radial shaft primary ossification center grows proportionately in length, transverse dimensions and volume, and quadratically in its projection surface area. The obtained numerical findings of the radial shaft ossification center are considered age-specific reference of relevance in both the estimation of fetal ages and the diagnostic process of congenital defects.

Fasciola Hepatica Isolates Induce Different Immune Responses in Unmaturated Bovine Macrophages.

INTRODUCTION: Fasciola hepatica (liver fluke) is a parasite of great socioeconomic importance. A number of fluke isolates have been identified; however, to date the differences between the immunomodulatory properties of different parasite isolates have not been sufficiently investigated. The aim of this study was to explore differences between the immunomodulatory properties of two F. hepatica isolates using unmaturated bovine macrophages. MATERIAL AND METHODS: A cell line of bovine macrophages was stimulated with excretory/secretory products released by adult flukes from either a laboratory (Fh-WeyES) or wild (Fh-WildES) strain and subsequently subjected to microarray and ELISA analyses. Results: Both Fh-WeyES and Fh-WildES dampened the release of interleukin-10 by bovine macrophages, but only Fh-WildES dampened the release of proinflammatory tumour necrosis factor-α. Microarray analysis revealed that Fh-WildES down- and upregulated 90 and 18 genes, respectively, when compared to Fh-WeyES. CONCLUSION: The results indicated different impacts of the isolates on macrophages. A number of researchers use flukes obtained from local slaughterhouses for experiments. Our findings may explain some discrepancies between published results arising from parasite strain choice. The findings indicate that consideration should be given to the use of different strains, and open new and currently unexplored avenues in parasitology for controlling the parasite.

Quantitative anatomy of the primary ossification center in the fetal pubis bone.

PURPOSES: Skeletodysplasiae and hereditary dysostoses constitute a group of over 350 disorders of the skeletal system. Knowledge about development of the pubic primary ossification center may be useful in both determining the fetal stage and maturity, and for detecting congenital disorders. The present study was performed to quantitatively examine the pubic primary ossification center with respect to its linear, planar, and volumetric parameters. MATERIALS AND METHODS: Using methods of computed tomography (CT), digital-image analysis and statistics, the size of the pubic primary ossification center in 33 spontaneously aborted human fetuses (18 males and 15 females) aged 22-30 weeks was studied. RESULTS: With no sex and laterality differences, the best-fit growth dynamics for the pubic primary ossification center was modeled by the following functions: y = - 13.694 + 0.728 × age ± 0.356 for its sagittal diameter, y = - 3.350 + 0.218 × age ± 0.159 for its vertical diameter, y = - 61.415 + 2.828 × age ± 1.519 for its projection surface area, and y = - 65.801 + 3.173 × age ± 2.149 for its volume. CONCLUSIONS: The size of the pubic primary ossification center shows neither sex nor laterality differences. The growth dynamics of the vertical and sagittal diameters, projection surface area, and volume of the pubic ossification centers follow proportionately to fetal age. The obtained numerical findings of the pubic ossification center are considered age-specific reference data with clinical implications in the diagnostics of congenital defects.

The primary ossification of the human fetal ischium: CT, digital-image analysis, and statistics.

PURPOSES: Details concerning the normal growth of the pelvic girdle in the fetus are of importance in the early detection of congenital defects. This study was executed to quantitatively evaluate the primary ossification center of the ischium with relation to its linear, planar and volumetric parameters. MATERIALS AND METHODS: Using methods of CT, digital-image analysis, and statistics, geometrical dimensions of the ischium's primary ossification center in 42 spontaneously aborted human fetuses (21 ♂ and 21 ♀) aged 18-30 weeks were calculated. RESULTS: With no sex and laterality differences, the best fit growth dynamics for the ischium's primary ossification center were displayed by the following functions: y = - 10.045 + 0.742 × age ± 0.013 (R2 = 0.97) for its vertical diameter, y = - 5.212 + 0.385 × age ± 0.008 (R2 = 0.97) for its sagittal diameter, y = - 36.401 + 0.122 × (age)2 ± 45.534 (R2 = 0.96) for its projection surface area, and y = - 1052.840 + 368.470 × ln(age) ± 12.705 (R2 = 0.91) for its volume. CONCLUSIONS: Neither male-female nor right-left differences are found for any of the morphometric parameters of the ischium's primary ossification center. With relation to fetal ages in weeks, the ischium's primary ossification center grows proportionately in vertical and sagittal diameters, second-degree polynomially in projection surface area, and logarithmically in volume. The quantitative findings of the ischium's primary ossification center are considered age-specific reference data of relevance in the diagnostics of innate defects.

Morphometric study of the primary ossification center of the fibular shaft in the human fetus.

PURPOSES: Precise morphometric data on the development of ossification centers in human fetuses may be useful in the early detection of skeletal dysplasias associated with delayed ossification center development and mineralization. The present study was performed to quantitatively examine the primary ossification center of the fibular shaft with respect to its linear, planar and volumetric parameters. MATERIALS AND METHODS: Using methods of CT, digital-image analysis (Osirix 3.9 MD) and statistics (Student's t-test, Shapiro-Wilk, Fisher's test, Tukey's test, Kruskal-Wallis test, regression analysis), the size of the primary ossification center of the fibular shaft in 47 spontaneously aborted human fetuses (25 ♂ and 22 ♀) aged 17-30 weeks was studied. In each fetus, the assessment of linear dimensions (length, transverse diameters for: proximal end, middle part and distal end), projection surface area and volume of the fibular shaft ossification center was carried out. RESULTS: With no sex and laterality differences, the best fit growth dynamics for the primary ossification center of the fibular shaft was modelled by the following functions: y = - 13.241 + 1.567 × age ± 1.556 (R2 = 0.94) for its length, y = - 0.091 + 0.063 × age ± 0.073 (R2 = 0.92) for its proximal transverse diameter, y = - 1.201 + 0.717 × ln(age) ± 0.054 (R2 = 0.83) for its middle transverse diameter, y = - 2.956 + 1.532 × ln(age) ± 0.090 (R2 = 0.89) for its distal transverse diameter, y = - 69.038 + 4.699 × age ± 4.055 (R2 = 0.95) for its projection surface area, and y = - 126.374 + 9.462 × age ± 8.845 (R2 = 0.94) for its volume. CONCLUSIONS: The ossification center in the fibular shaft follows linear functions with respect to its length, proximal transverse diameter, projection surface area and volume, and natural logarithmic functions with respect to its middle and distal transverse diameters. The obtained morphometric data of the fibular shaft ossification center is considered normative for their respective prenatal weeks and may be of relevance in both the estimation of fetal age and the ultrasound diagnostics of congenital defects.

Three-dimensional growth of tibial shaft ossification in the human fetus: a digital-image and statistical analysis.

PURPOSES: Tibial shaft ossification in terms of its size and growth may be criticalin describing both the fetal stage and maturity, and in identifying innate disorders. The present study was executed to quantitatively assess ossification of the tibial shaft, taking its morphometric linear, planar and volumetric parameters into account. MATERIALS AND METHODS: With the use of methods of CT, digital-image analysis and statistics, the evolutionof tibial shaft ossification in 47 spontaneously aborted human fetuses at the age of 17-30 weeks was studied. RESULTS: Without any male-female and right-left morphometric differences, the best fit growth dynamics fortibial shaft ossification was modelled by the following functions: y = 5.312 + 0.034 × (age)2 ± 0.001 (R2 = 0.89) for its length, y = - 2.855 + 0.307 × age ± 0.009 (R2 = 0.96) for its proximal transverse diameter, y = - 0.758 + 0.153 × age ± 0.005 (R2 = 0.88) for its middle transverse diameter, y = - 1.844 + 0.272 × age ± 0.09 (R2 = 0.90) for its distal transverse diameter, y = - 40.263 + 0.258 × (age)2 ± 0.007 (R2 = 0.94) for its projection surface area, and y = - 287.996 + 1.186 × (age)2 ± 0.037 (R2 = 0.92) for its volume. The femoral-to-tibial ossification length ratio was 1.15 ± 0.1. CONCLUSIONS: The size of tibial shaft ossification displays neither sex nor laterality differences. Tibial shaft ossification follows quadratic functions with respect to its length, projection surface area and volume, and linear functions with respect to its proximal, middle and distal transverse diameters. The obtained morphometric data of tibial shaft ossification are considered normative age-specific references of relevance in both the estimation of fetal ages and the ultrasound diagnostics of congenital defects.

Quantitative anatomy of the ulna's shaft primary ossification center in the human fetus.

PURPOSE: There has been little information in the medical literature regarding the growing ulna in the human fetus, though such knowledge appears to be potentially useful in diagnosing skeletal dysplasias, characterized by a disrupted or completely halted growth of the fetus. Therefore, longitudinal measurements of long bones are extremely conducive in assessing both pregnancy and fetal anatomy. MATERIALS AND METHODS: Using methods of CT, digital-image analysis and statistics, the size of the ulna's shaft primary ossification center in 48 (26 males and 22 females) spontaneously aborted human fetuses aged 17-30 weeks was studied. RESULTS: With no sex differences, the best fit growth dynamics for the ulna's shaft primary ossification center was modeled by the following functions: y = - 8.476 + 1.561 × age ± 0.019 for its length, y = - 2.961 + 0.278 × age ± 0.016 for its proximal transverse diameter, y = - 0.587 + 0.107 × age ± 0.027 for its middle transverse diameter, y = - 2.865 + 0.226 × age ± 0.295 for its distal transverse diameter, y = - 50.758 + 0.251 × (age)2 ± 0.016 for its projection surface area, and y = - 821.707 + 52.578 × age ± 0.018 ± 102.944 for its volume. CONCLUSIONS: The morphometric characteristics of the ulna's shaft primary ossification center show neither sex nor bilateral differences. The ulna's shaft primary ossification center grows linearly with respect to its length, transverse dimensions and volume, and follows a quadratic function with respect to its projection surface area. The obtained morphometric data of the ulna's shaft primary ossification center is considered normative for respective prenatal weeks and may be of relevance in both the estimation of fetal ages and the diagnostic process of congenital defects.