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Voprosy Virusologii Jul 2023Following the successful eradication of smallpox, mass vaccination against this disease was discontinued in 1980. The unvaccinated population continues to be at risk of...
INTRODUCTION
Following the successful eradication of smallpox, mass vaccination against this disease was discontinued in 1980. The unvaccinated population continues to be at risk of infection due to military use of variola virus or exposure to monkeypox virus in Africa and non-endemic areas. In cases of these diseases, rapid diagnosis is of great importance, since the promptness and effectiveness of therapeutic and quarantine measures depend on it. The aim of work is to develop a kit of reagents for enzyme-linked immunosorbent assay (ELISA) for fast and highly sensitive detection of orthopoxviruses (OPV) in clinical samples.
MATERIALS AND METHODS
The efficiency of virus detection was evaluated by single-stage ELISA in the cryolisate of CV-1 cell culture samples infected with vaccinia, cowpox, rabbitpox, and ectromelia viruses, as well as in clinical samples of infected rabbits and mice.
RESULTS
The method of rapid ELISA was shown to allow the detection of OPV in crude viral samples in the range of 5.0 1025.0 103 PFU/ml, and in clinical samples with a viral load exceeding 5 103 PFU/ml.
CONCLUSIONS
The assay involves a minimum number of operations and can be performed within 45 minutes, which makes it possible to use it in conditions of a high level of biosecurity. Rapid ELISA method was developed using polyclonal antibodies, which significantly simplifies and reduces the cost of manufacturing a diagnostic system.
Topics: Rabbits; Animals; Mice; Orthopoxvirus; Vaccinia virus; Variola virus; Ectromelia virus; Enzyme-Linked Immunosorbent Assay
PubMed: 37436415
DOI: 10.36233/0507-4088-178 -
Cells Dec 2023Conventional dendritic cells (cDCs) are innate immune cells that play a pivotal role in inducing antiviral adaptive immune responses due to their extraordinary ability...
Conventional dendritic cells (cDCs) are innate immune cells that play a pivotal role in inducing antiviral adaptive immune responses due to their extraordinary ability to prime and polarize naïve T cells into different effector T helper (Th) subsets. The two major subpopulations of cDCs, cDC1 (CD8α in mice and CD141 in human) and cDC2 (CD11b in mice and CD1c in human), can preferentially polarize T cells toward a Th1 and Th2 phenotype, respectively. During infection with ectromelia virus (ECTV), an orthopoxvirus from the family, the timing and activation of an appropriate Th immune response contributes to the resistance (Th1) or susceptibility (Th2) of inbred mouse strains to the lethal form of mousepox. Due to the high plasticity and diverse properties of cDC subpopulations in regulating the quality of a specific immune response, in the present study we compared the ability of splenic cDC1 and cDC2 originating from different ECTV-infected mouse strains to mature, activate, and polarize the Th immune response during mousepox. Our results demonstrated that during early stages of mousepox, both cDC subsets from resistant C57BL/6 and susceptible BALB/c mice were activated upon in vivo ECTV infection. These cells exhibited elevated levels of surface MHC class I and II, and co-stimulatory molecules and showed enhanced potential to produce cytokines. However, both cDC subsets from BALB/c mice displayed a higher maturation status than that of their counterparts from C57BL/6 mice. Despite their higher activation status, cDC1 and cDC2 from susceptible mice produced low amounts of Th1-polarizing cytokines, including IL-12 and IFN-γ, and the ability of these cells to stimulate the proliferation and Th1 polarization of allogeneic CD4 T cells was severely compromised. In contrast, both cDC subsets from resistant mice produced significant amounts of Th1-polarizing cytokines and demonstrated greater capability in differentiating allogeneic T cells into Th1 cells compared to cDCs from BALB/c mice. Collectively, our results indicate that in the early stages of mousepox, splenic cDC subpopulations from the resistant mouse strain can better elicit a Th1 cell-mediated response than the susceptible strain can, probably contributing to the induction of the protective immune responses necessary for the control of virus dissemination and for survival from ECTV challenge.
Topics: Humans; Animals; Mice; Mice, Inbred C57BL; Ectromelia, Infectious; Poxviridae Infections; Cytokines; Dendritic Cells
PubMed: 38201217
DOI: 10.3390/cells13010013 -
Anatomical Record (Hoboken, N.J. : 2007) Jan 2022The congenitally shortened limb (CSL) with fibular deficiency or absence has historically been graded by plain radiography, while associated cartilaginous and arterial... (Review)
Review
The congenitally shortened limb (CSL) with fibular deficiency or absence has historically been graded by plain radiography, while associated cartilaginous and arterial soft tissue anomalies have been comparatively neglected. Consistent pathological evidence of remnant cartilaginous bodies in place of the fibula presupposes earlier existence of a preformed cartilaginous template of the fibula. In complete fibular radiographic absences, often associated with midline metatarsal deficiencies, the two usual nutrient arteries to the fibula fail to form, as they normally would have, around the (16-18 mm stage) sixth embryonic week. The histopathology of fallow persisting fibular anlagen, in association with missing arteries and retained primitive arteries, suggests the anlage is a dystrophic, but otherwise normally prefigured, cartilaginous scaffold of the fibula. Thus, the widely employed term absent fibula, which has been grounded in plain radiography, is a misnomer. Additionally, since the metatarsals missing in congenitally shortened limb are midline, the related term, fibular hemimelia, is similarly inaccurate. A new taxonomy, based on embryological principles rather than radiographic appearance alone, will promote limb dystrophism as a more accurate term combining arrested embryonic vascular development and congenitally shortened limb of the lower extremity.
Topics: Ectromelia; Embryonic Development; Fibula; Humans; Lower Extremity; Radiography
PubMed: 33773063
DOI: 10.1002/ar.24628 -
Experimental and Therapeutic Medicine Apr 2021Cranioectodermal dysplasia (CED) or Sensenbrenner syndrome is a very rare autosomal-recessive disease that is characterized by craniofacial, skeletal and ectodermal...
Cranioectodermal dysplasia (CED) or Sensenbrenner syndrome is a very rare autosomal-recessive disease that is characterized by craniofacial, skeletal and ectodermal abnormalities. The proteins encoded by six CED-associated genes are members of the intraflagelline transport (IFT) system, which serves an essential role in the assembly, maintenance and function of primary cilia. The current study identified compound novel heterozygous (NM_052985.3) variants in a male Chinese infant with CED. The latter variant changes the length of the protein and may result in the partial loss-of-function of IFT122. With the simultaneous presence of frameshift and stop-loss variants, the patient manifested typical CED with fine and sparse hair, macrocephaly, dysmorphic facial features and upper limb phocomelia. A number of unusual phenotypic characteristics were additionally observed and included postaxial polydactyly of both hands and feet. The molecular confirmation of CED in this patient expands the CED-associated variant spectrum of in CED, while the manifestation of CED in this patient provides additional clinical information regarding this syndrome. Moreover, the two variants identified in the proband provide a novel perspective into the phenotypes caused by different combinations of variants.
PubMed: 33717254
DOI: 10.3892/etm.2021.9742 -
Journal of Feline Medicine and Surgery Oct 2023The present study aimed to determine the inheritance pattern and genetic cause of congenital radial hemimelia (RH) in cats.
OBJECTIVES
The present study aimed to determine the inheritance pattern and genetic cause of congenital radial hemimelia (RH) in cats.
METHODS
Clinical and genetic analyses were conducted on a Siamese cat family (n = 18), including two siblings with RH. Radiographs were obtained for the affected kittens and echocardiograms of an affected kitten and sire. Whole genome sequencing was completed on the two cases and the parents. Genomic data were compared with the 99 Lives Cat Genome data set of 420 additional domestic cats with whole genome and whole exome sequencing data. Variants were considered as homozygous in the two cases of the siblings with RH and heterozygous in the parents. Candidate variants were genotyped by Sanger sequencing in the extended pedigree.
RESULTS
Radiographs of the female kitten revealed bilateral absence of the radii and bowing of the humeri, while the male kitten showed a dysplastic right radius. Echocardiography suggested the female kitten had restrictive cardiomyopathy with a positive left atrial-to-aortic root ratio (LA:Ao = 1.83 cm), whereas hypertrophic cardiomyopathy was more likely in the sire, showing diastolic dysfunction using tissue Doppler imaging (59.06 cm/s). Twenty-two DNA variants were unique and homozygous in the affected kittens and heterozygous in the parents. Seven variants clustered in one chromosomal region, including two frameshift variants in () and five variants in , ( ), including a missense and an in-frame deletion.
CONCLUSIONS AND RELEVANCE
The present study suggested an autosomal recessive mode of inheritance with variable expression for RH in the Siamese cat family. Candidate variants for the phenotype were identified, implicating their roles in bone development. These genes should be considered as potentially causal for other cats with RH. Siamese cat breeders should consider genetically testing their cats for these variants to prevent further dissemination of the suspected variants within the breed.
Topics: Female; Male; Cats; Animals; Ectromelia; Cardiomyopathies; Risk Factors; Cardiomyopathy, Hypertrophic; Humerus; Cat Diseases
PubMed: 37791865
DOI: 10.1177/1098612X231193557 -
Journal of Virology Feb 2023Receptor-interacting protein kinase 3 (RIPK3) and mixed lineage kinase domain-like pseudokinase (MLKL) are proteins that are critical for necroptosis, a mechanism of...
Receptor-interacting protein kinase 3 (RIPK3) and mixed lineage kinase domain-like pseudokinase (MLKL) are proteins that are critical for necroptosis, a mechanism of programmed cell death that is both activated when apoptosis is inhibited and thought to be antiviral. Here, we investigated the role of RIPK3 and MLKL in controlling the Orthopoxvirus ectromelia virus (ECTV), a natural pathogen of the mouse. We found that C57BL/6 (B6) mice deficient in RIPK3 () or MLKL () were as susceptible as wild-type (WT) B6 mice to ECTV lethality after low-dose intraperitoneal infection and were as resistant as WT B6 mice after ECTV infection through the natural footpad route. Additionally, after footpad infection, mice, but not mice, endured lower viral titers than WT mice in the draining lymph node (dLN) at three days postinfection and in the spleen or in the liver at seven days postinfection. Despite the improved viral control, mice did not differ from WT mice in the expression of interferons or interferon-stimulated genes or in the recruitment of natural killer (NK) cells and inflammatory monocytes (iMOs) to the dLN. Additionally, the CD8 T-cell responses in and WT mice were similar, even though in the dLNs of mice, professional antigen-presenting cells were more heavily infected. Finally, the histopathology in the livers of and WT mice at 7 dpi did not differ. Thus, the mechanism of the increased virus control by mice remains to be defined. The molecules RIPK3 and MLKL are required for necroptotic cell death, which is widely thought of as an antiviral mechanism. Here we show that C57BL/6 (B6) mice deficient in RIPK3 or MLKL are as susceptible as WT B6 mice to ECTV lethality after a low-dose intraperitoneal infection and are as resistant as WT B6 mice after ECTV infection through the natural footpad route. Mice deficient in MLKL are more efficient than WT mice at controlling virus loads in various organs. This improved viral control is not due to enhanced interferon, natural killer cell, or CD8 T-cell responses. Overall, the data indicate that deficiencies in the molecules that are critical to necroptosis do not necessarily result in worse outcomes following viral infection and may improve virus control.
Topics: Animals; Mice; Ectromelia virus; Ectromelia, Infectious; Interferons; Mice, Inbred C57BL; Necroptosis; Protein Kinases; Receptor-Interacting Protein Serine-Threonine Kinases
PubMed: 36651749
DOI: 10.1128/jvi.01945-22 -
Birth Defects Research Nov 2022Sirenomelia is a congenital malformation of the lower body characterized by a single midline lower limb and severe urogenital and gastrointestinal malformations....
BACKGROUND
Sirenomelia is a congenital malformation of the lower body characterized by a single midline lower limb and severe urogenital and gastrointestinal malformations. Sirenomelia is rare (estimated incidence of approximately 1/100,000) and usually lethal in the perinatal period.
CASE
A 2,042 g Japanese male infant, one of monochorionic monoamniotic twins, was born at 34 weeks of gestation by elective caesarean section. Sirenomelia was prenatally diagnosed. Single midline lower limb, bilateral dysplastic kidneys, an omphalomesenteric fistula, colon atresia, imperforate anus, indiscernible genital structures, and myelomeningocele were detected at birth. The amniotic fluid volume was normal throughout the pregnancy course, which led to appropriate lung maturation of the twin with sirenomelia. Although renal replacement therapy was initiated soon after birth, stable peritoneal dialysis was difficult because of the limited intraperitoneal space, and the infant frequently developed peritonitis. He died of sudden cardiorespiratory arrest at 6 months of age. Postmortem examination showed bilateral dysplastic kidneys, agenesis of the ureters and urinary bladder, abnormal branching and agenesis of the distal colon, bilateral inguinal hernias, and small testes.
CONCLUSION
Infants with sirenomelia, even those with end-stage kidney disease at birth, may survive if they have a stable cardiorespiratory status at birth and renal replacement therapy is appropriately initiated.
Topics: Female; Humans; Infant; Infant, Newborn; Male; Pregnancy; Amnion; Anus, Imperforate; Cesarean Section; Ectromelia; Twins, Monozygotic; Treatment Outcome
PubMed: 35437955
DOI: 10.1002/bdr2.2016 -
Ultrasound in Obstetrics & Gynecology :... Apr 2023
Topics: Humans; Amniotic Fluid; Ectromelia; Kidney; Kidney Diseases; Congenital Abnormalities
PubMed: 36173397
DOI: 10.1002/uog.26076 -
Prenatal Diagnosis Apr 2020To determine the key sonographic features for the diagnosis of sirenomelia in the first trimester of pregnancy. (Review)
Review
OBJECTIVE
To determine the key sonographic features for the diagnosis of sirenomelia in the first trimester of pregnancy.
METHODS
Cases of sirenomelia from several prenatal diagnosis centers were retrospectively identified and reviewed. The diagnosis was established through the detection of fused lower limbs. Additional sonographic findings were also noted.
RESULTS
A total of 12 cases were collected. The most striking sonographic finding was the detection of malformed lower limbs, which appeared to be fused and in an atypical position. Nuchal translucency thickness was mildly increased in three cases (25%). An abdominal cyst, representing the dilated blind-ending bowel, was noted in seven cases (58%). Color flow imaging detected a single umbilical artery in six cases (50%) and the associated intra-abdominal vascular anomalies in three cases (25%). No cases of aneuploidy were detected. The pregnancy was terminated in nine cases (75%) and intrauterine demise occurred in the remaining three cases (25%).
CONCLUSIONS
The sonographic detection of abnormal lower limbs or an intra-abdominal cyst located laterally during the first-trimester scan may be warning signs of sirenomelia. This should prompt a detailed examination of the fetal lower body and intra-abdominal anatomy, including the main abdominal vessels, in order to look for additional confirmatory findings.
Topics: Abortion, Induced; Adult; Digestive System Abnormalities; Ectromelia; Female; Fetal Death; Humans; Intestines; Lower Extremity Deformities, Congenital; Nuchal Translucency Measurement; Pregnancy; Pregnancy Trimester, First; Single Umbilical Artery; Ultrasonography, Doppler, Color; Ultrasonography, Prenatal; Vascular Malformations
PubMed: 32040213
DOI: 10.1002/pd.5667 -
International Journal of Environmental... Apr 2021This study investigates the acceptability, bioethical justification, and determinants of the provision of intensive care to extremely preterm or ill neonates among...
BACKGROUND
This study investigates the acceptability, bioethical justification, and determinants of the provision of intensive care to extremely preterm or ill neonates among healthcare professionals serving in NICUs in Greek hospitals.
METHODS
Healthcare professionals (71 physicians, 98 midwives, and 82 nurses) employed full-time at all public Neonatal Intensive Care Units (NICUs) ( = 17) in Greece were asked to report their potential behavior in three clinical scenarios.
RESULTS
The majority of healthcare professionals would start and continue intensive care to (a) an extremely preterm neonate, (b) a full-term neonate with an unfavorable prognosis, and (c) a neonate with complete phocomelia. In cases (a) and (b), midwives and nurses compared to physicians ( = 0.009 and = 0.004 in scenarios (a) and (b), respectively) and health professionals ascribing to the quality-of-life principle compared to those ascribing to the intrinsic value of life ( = 0.001 and = 0.01 scenarios (a) and (b) respectively), tend towards withholding or withdrawing care. Religion plays an important role in all three scenarios ( = 0.005, = 0.017 and = 0.043, respectively).
CONCLUSIONS
Understanding healthcare professionals' therapeutic intensiveness in the face of NICU ethical dilemmas can improve NICU policies, support strategies, and, consequently, the quality of neonatal intensive care.
Topics: Attitude of Health Personnel; Decision Making; Female; Greece; Humans; Infant, Newborn; Intensive Care Units, Neonatal; Midwifery; Physicians; Pregnancy
PubMed: 33918554
DOI: 10.3390/ijerph18083938