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Comparison of polymerase chain reaction and bacteriological culture for the diagnosis of sheep brucellosis using aborted fetus samples.

PCR assay has been shown to be a promising option for the diagnosis of brucellosis. However, few studies have been performed with field samples in order to evaluate the assay as a diagnostic tool. In this study, routine use of a species-specific PCR assay previously developed for the identification of Brucella cultures was assessed for the detection of Brucella DNA directly from the stomach contents of aborted sheep fetuses. The assay is based on the insertion sequence IS711 in the Brucella chromosome. In the study, during 3 successive lambing seasons (1998-1999, 1999-2000 and 2000-2001) 126 aborted fetus samples each from different flocks and locations were examined. Brucella strains were isolated from 39 (31%) of the samples and all of the strains were identified as Brucella melitensis by biochemical characteristics, agglutination with monospecific A and M sera and PCR. Thirty-seven of 39 B. melitensis isolates were biotyped as biotype 3, and 2 isolates as biotype 1. From 38 of 39 culture positive fetal stomach contents B. melitensis-specific DNA was detected by PCR. PCR was found negative in all of the culture negative samples. Compared with culture, sensitivity and specificity of PCR were determined as 97.4 and 100%, respectively. The results indicate that this PCR procedure has a potential for use in routine diagnosis of sheep brucellosis.

Aborted Fetus↗

Thymic abnormalities in fetuses aborted from human immunodeficiency virus type 1 seropositive women.

Pathological abnormalities of the thymus were found in 3 of 37 fetuses aborted from human immunodeficiency virus (HIV)-infected mothers. These lesions were located predominantly in the thymic cortex, which contains mostly immature lymphocytes. Areas of focal lymphocyte depletion were infiltrated with CD4+ macrophages and were associated with abnormalities of the epithelial stromal network. No evidence of extensive HIV infection in any of the 37 thymuses was detected by either immunofluorescence or in situ hybridization techniques, although rare cells that expressed HIV antigens were found in 3 fetuses. Although less extensive, this thymic fetopathy was similar to that described in postnatal acquired immunodeficiency syndrome thymuses, strongly suggesting that the lesions were related to HIV infection. Thymic fetopathy might represent the initial injury to the lymphoid system in HIV-infected infants in whom early and severe immunosuppression develops.

Abortion, Induced↗

[Prevalence and association of porcine circovirus type 2 (PCV2), porcine parvovirus (PPV) and porcine reproductive and respiratory syndrome virus (PRRSV) in aborted fetuses, mummified fetuses, stillborn and nonviable neonatal piglets].

Porcine circovirus type 2 (PCV2) seems to cause reproductive failure in sows not only in experimental studies. A retrospective study was made with a total of 252 aborted fetuses, mummified fetuses, stillborn and nonviable neonatal piglets to determine the presence of PCV2, porcine parvovirus (PPV) and porcine respiratory and reproductive syndrome virus (PRRSV) by PCR. PCV2 was found in all stages of gestation in 27.1 percent of samples examined. A statistically significant association could be shown between the detection of PCV2 and PRRSV. However, no significant association was seen between the detection of PCV2 and PPV and between PPV and PRRSV.

Abortion, Veterinary↗

Comparison of virus isolation, reverse transcription-polymerase chain reaction, immunohistochemistry, and in situ hybridization for the detection of porcine reproductive and respiratory syndrome virus from naturally aborted fetuses and stillborn piglets.

Virus isolation, reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry, and in situ hybridization methods were compared for the detection of porcine reproductive and respiratory syndrome virus (PRRSV). Seven aborted fetuses and 6 stillborn piglets naturally infected with PRRSV were used in the study. Viral antigen and viral nucleic acid were detected in macrophages and dendritic cells in the spleen, tonsil, lymph nodes, and thymus; in macrophages of liver, heart, and lung; and in endothelial cells and myocytes of the heart. Viral antigen and viral nucleic acid were most consistently detected in the spleen. Of the 13 samples, 6 were positive for PRRSV by all 4 techniques. Four (31%) samples were positive for PRRSV by RT-PCR, in situ hybridization, and virus isolation. Two (15%) samples were positive for PRRSV by virus isolation, RT-PCR, and in situ hybridization. One (8%) was positive for PRRSV by virus isolation and RT-PCR. The RT-PCR identified the presence of PRRSV more frequently than the other methods. However, when only formalin-fixed tissues are submitted, immunohistochemistry and in situ hybridization would be useful methods for the detection of PRRSV antigen and nucleic acid.

Abortion, Veterinary↗

[Complete genome sequence of 84FLi, a Hantaan virus strain isolated from the liver of fetus aborted by a pregnant women with hemmorrhagic fever with renal syndrome].

OBJECTIVE: To study the complete genome sequence of the Chinese Hantaan virus vaccine strain 84FLi and learn about its molecular characters. METHODS: The virus strain 84FLi was isolated from the liver of a fetus aborted by pregnant women with hemorrhagic fever with renal syndrome. The cDNAs of L M and S segments were amplified fragment by fragment using RT-PCR. The purified PCR products were sequenced directly or cloned into pMD18-T vector and then sequenced. RESULTS: The complete genome of strain 84FLi was composed of L ( 6 533 bp) M (3 616 bp) and S (1 688 bp) coding 2151 1135 and 429 amino acids respectively. The entire sequence composition was 3830A 2050C 2510G and 3447T the GC and AT contents were 38.52% and 61.48%. Homology analysis showed that the homologies of 84 FLi S segment nucleotide sequences with strain RG9 (isolated in Guangzhou) and strain Chen4 (isolated in Anhui) were 99.6%. There were 83.7% 84.0% and 87.2% nucleotide sequences homology with the three segments of Hantaan virus foreign standard strain 76-118 while the amino acids sequences homology with those of 76-118 were 97.5% 96.0% and 97.9% respectively. CONCLUSION: The strain 84FLi is highly related to other Chinese Hantaan virus isolates and is in the same subtype with the other two Chinese isolates RG9 and Chen4.

Amino Acid Sequence↗

Identification and PCR-restriction fragment length polymorphism analysis of a variant of the Ibaraki virus from naturally infected cattle and aborted fetuses in Japan.

One hundred fourteen field isolates of the Ibaraki virus (IBAV), a member of the epizootic hemorrhagic disease virus serotype 2 (EHDV-2), were isolated from blood samples of affected and apparently healthy cattle and Culicoides biting midges and from blood samples of dams and internal organs of aborted fetuses during an outbreak of Ibaraki disease in the southern part of Japan in 1997. In this outbreak, 242 cattle showed typical symptoms of the disease, and several hundred dams had miscarriages or stillbirths. The viruses that induced typical Ibaraki disease and reproductive problems among cattle were identical and were antigenically closely related to but distinct from previous isolates of IBAV and EHDV-2. The virus was considered to be a putative agent of this outbreak. Reverse transcription-PCR based on segment 3 of the RNA genome of EHDV-2 and restriction fragment length polymorphism analysis of the PCR products were conducted to compare the genomes of the viruses. The results suggested that the virus isolated in 1997 was a variant of IBAV and might be exotic.

Abortion, Veterinary↗

Eye findings in 8 children and a spontaneously aborted fetus with RSH/Smith-Lemli-Opitz syndrome.

We evaluate the ophthalmologic findings in 8 children with RSH/Smith-Lemli-Opitz syndrome (SLOS) and document abnormal concentrations of cholesterol and cholesterol precursors in the ocular tissues in a case of SLOS. The most common ophthalmologic finding was blepharoptosis, which was found in 6 of 8 patients, with the severity ranging from mild to moderate. None of the patients in the present study demonstrated cataracts; none had amblyopia from blepharoptosis. One patient had a right hypertropia with overaction of the inferior oblique muscle. This patient also had optic atrophy and a second patient had bilateral optic nerve hypoplasia. The importance of these findings to the visual function remains to be defined. Sterol analysis from ocular tissues of an aborted fetus with SLOS showed increased 7- and 8-dehydrocholesterol and a low cholesterol concentration in the retinal pigment epithelium, lens, cornea, and sclera. Routine ophthalmologic examination is indicated in SLOS because of the high incidence of abnormalities, most likely due to the abnormal synthesis of cholesterol and cholesterol precursors in the ocular tissues of these patients, as evidenced by sterol analysis of the ocular tissues in a case of SLOS.

Child, Preschool↗

[Molecular-cytogenetic study of the aborted fetuses in women with reproductive function disorders].

It is known that the frequency of chromosomal abnormalities among spontaneous miscarriages of the first trimester of pregnancy makes 50-60%. Research of karyotypes of chorionic villus cells of miscarriages has been conducted by combining the standard cytogenetic method and the FISH analysis on interphase nuclei of centromeric specific DNA samples by the tests to the chromosomes 13/21, 14/22, 15, 16, 18, X, Y. The described complex approach can be successfully applied for effective identification ofchromosomal abnormalities in the material of spontaneous miscarriages. The results specify the necessity of careful study of genomes of matrimonial pairs with the usual unmaturing in anamnesis and especially before treatment by IVF methods.

Aborted Fetus↗

Campylobacteriosis in an aborted equine fetus.

Abortion caused by Campylobacter fetus subsp fetus was diagnosed in a 7-month-old equine fetus. The fetus was small for its gestational age. Macroscopically, the proximal portion of the small intestine was hemorrhagic and its wall was thick. Histologically, the Brunner glands were distended with neutrophils, and the submucosa was thick, owing to fluid accumulation and/or cellular infiltrates. Curved bacteria were observed in the Brunner glands and intestinal glands. Campylobacter fetus subsp fetus was isolated from stomach contents, liver, and lungs, and was detected by dark-field microscopic examination of ocular fluid and stomach contents. Placenta was not available for examination.

Abortion, Veterinary↗

First Portuguese isolate of Neospora caninum from an aborted fetus from a dairy herd with endemic neosporosis.

Neospora caninum was isolated from the brain of an aborted 4-month-old fetus from a dairy cow herd with endemic neosporosis in Porto, Portugal. The fetal brain homogenate was inoculated interperitoneally first into outbred Swiss Webster mice given dexamethasone and then the peritoneal exudates from these mice was co-inoculated with mouse sarcoma cells in the peritoneal cavity of mice given dexamethasone. N. caninum tachyzoites were seen in peritoneal exudate of the second passage. Tachyzoites from the peritoneal exudate reacted positively with anti-N. caninum antibodies and not with anti-Toxoplasma gondii antibodies and contained N. caninum specific DNA. This Portuguese isolate of N. caninum has been successfully maintained in cell culture. The dam of the aborted fetus had an antibody titer of 1:10240 in the Neospora agglutination test (NAT). Antibodies to N. caninum were found in 76 of 106 cows from this herd in titers of 1:40 in 31, 1:80 in 22, > or =1:160 or more in 23 in the Neospora agglutination test. This is the first isolation of a viable N. caninum-like parasite from any host in Portugal.

Abortion, Veterinary↗

Defect in dorso-ventral patterning, asplenia, and conotruncus in a spontaneously aborted fetus.

We describe a very unusual combination, and previously unreported, of malformations in an 18-week, spontaneously aborted male fetus. The fetus had a reversed dorsoventral positioning of the head and upper limbs relative to the body axis with the head and both upper limbs directed dorsally, and an abrupt rotation of the vertebral bones at the level of CZ The fetus also had asplenia, single ventricle, and conotruncus. The fetus also had flexion deformities at the wrist, reduction deformity of the left second digit, anomalies in ossification of the bones of the left hand, and bilateral talipes calcaneovalgus. The major malformations in this fetus were all blastogenetic in origin and consisted of dorsoventral patterning defect involving structures cranial to C7, a laterality, and a septation defect of the ventricle and outflow tract of the heart. The defects are interpreted as being the result of abnormal coordination of the molecular signaling involved in dorsoventral axis formation and laterality of the limbs and trunk, and possibly also in cardiac septation.

Abnormalities, Multiple↗