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11 pages, 445 KiB  
Article
Likelihood of Preterm Birth in Patients After Antenatal Corticosteroid Administration in Relation to Diagnosis and Confounding Risk Factors: A Retrospective Cohort Study
by Jan Pauluschke-Fröhlich, Richard Berger, Harald Abele, Claudia F. Plappert and Joachim Graf
Healthcare 2025, 13(1), 87; https://doi.org/10.3390/healthcare13010087 - 6 Jan 2025
Viewed by 69
Abstract
Background: In the case of threatened preterm birth (PTB) before the 34th week of pregnancy, the application of antenatal corticosteroids (ACSs) for the maturation of the fetal lung is a standard procedure in perinatal medicine. Common diagnoses for ACS use in pregnancy [...] Read more.
Background: In the case of threatened preterm birth (PTB) before the 34th week of pregnancy, the application of antenatal corticosteroids (ACSs) for the maturation of the fetal lung is a standard procedure in perinatal medicine. Common diagnoses for ACS use in pregnancy are the preterm rupture of membranes (PPROMs), placental bleeding, premature labor, preeclampsia, oligohydramnios, amniotic infection syndrome (AIS), and cervical insufficiency. The aim of this study was to investigate whether the current diagnosis, which results in ACS, and the patient’s risk factors influence the risk of PTB events. Methods: The data of all affected women were extracted, who were hospitalized in 2016 due to a threatening PTB and administered corticosteroids in the German federal state Rhineland-Palatinate (n = 1544), so the study was conducted as a retrospective cohort trial. Frequency analyses, Friedman tests, Chi square tests, logistic regressions, Spearman correlation, and moderation analysis were performed to determine the Odds ratio (OR) for PTB in ACS patients in relation to diagnosis and risk factors. Results: Only 60% of all patients with PTB risk delivered prematurely, whereby patients with different diagnoses differ in terms of the PTB risk; the highest risk could be found in AIS (OR = 16.12) or preeclampsia (OR = 5.46). For prior PTB, stillbirth, or abortion, there is a moderation effect (based on the confounders), while multiple pregnancies influence the PTB risk irrespective of diagnosis (OR = 6.45). In the whole collective, the OR for PTB was 6.6 in relation to all pregnant women in Germany. Conclusions: A higher risk of PTB could be found in patients with a diagnosis of AIS, preeclampsia, as well as in multiple pregnancies. Prior PTB, stillbirth, or abortion act as a relevant confounder. Full article
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<p>Distribution of diagnoses in the study population, responsible for the induction of lung maturation, <span class="html-italic">n</span> = 1544. AIS = amniotic infection syndrome; PPROM = preterm premature rupture of membranes.</p>
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19 pages, 1544 KiB  
Article
First Molecular and Phylogenetic Characterization of Equine Herpesvirus-1 (EHV-1) and Equine Herpesvirus-4 (EHV-4) in Morocco
by Zineb El Brini, Ann Cullinane, Marie Garvey, Ouafaa Fassi Fihri, Siham Fellahi, Farid Amraoui, Chafiqa Loutfi, Ghizlane Sebbar, Romain Paillot and Mohammed Piro
Animals 2025, 15(1), 102; https://doi.org/10.3390/ani15010102 - 5 Jan 2025
Viewed by 243
Abstract
This study aimed to investigate the molecular prevalence and genetic characterization of EHV-1 and EHV-4 in equid populations in Morocco. A total of 154 equids (114 horses, 9 donkeys, and 31 mules) were sampled, with nasal swabs and tissue samples subjected to multiplex [...] Read more.
This study aimed to investigate the molecular prevalence and genetic characterization of EHV-1 and EHV-4 in equid populations in Morocco. A total of 154 equids (114 horses, 9 donkeys, and 31 mules) were sampled, with nasal swabs and tissue samples subjected to multiplex real-time PCR for the detection of EHV-1 and EHV-4. Additionally, an isolate from the tissue of an aborted horse fetus was included in the analysis. A subset of EHV-positive samples underwent virus isolation followed by whole-genome sequencing. PCR assays revealed that 42 samples (27%) tested positive for EHV-4, while only 3 samples (1.94%) were positive for EHV-1. Attempts to isolate the virus from EHV-4-positive samples were unsuccessful. However, virus isolation was successful in an EHV-1-positive nasopharyngeal sample from a donkey. Phylogenetic and molecular characterization reclassified the EHV-1 isolated from the donkey as an EHV-8. Meanwhile, the EHV-1 isolated from the aborted fetal tissue was classified as a clade 1 EHV-1 virus. This study is the first to report the molecular prevalence and genetic characterization of EHV-1 and EHV-4 in equid populations in Morocco, providing valuable insights into the distribution and genetic diversity of these viruses in the region. Full article
(This article belongs to the Section Equids)
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<p>Cytopathogenic effect of EHV-1 (the respiratory donkey) on RK-13 cells, showing syncytium formation (arrows) and ballooning degeneration (arrowheads). The left image (40 × magnification) provides an overview of the cytopathic effects, while the right image (200 × magnification) offers a detailed view of the cellular alterations.</p>
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<p>Alignment of the predicted amino acid sequence of ORF30 polymerase gene for EHV-1 and EHV-8 viruses including the two Moroccan viruses. Amino acid positions are indicated above the alignment. Identical residues are indicated with a dot. Amino acid residue D752 corresponds to the proposed marker of neuropathogenicity in EHV-1 (highlighted in yellow). The Moroccan strains are framed in red.</p>
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<p>Maximum-likelihood phylogenetic tree (scale bar approximately 0.02 substitutions per site), indicating greater evolutionary distance or divergence among equid alphaherpesvirus strains, including the two Moroccan viruses sequenced in this study (highlighted in bold). The analysis included 50 genome sequences, and bootstrap values calculated from 1000 replicates are shown at major nodes.</p>
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<p>Phylogenetic analysis of EHV-1 genomes from 13 distinct UL clades (numbered 1 to 13, with each clade represented by a different color), including the Moroccan strain EHV1/MA/2010/21 sequenced in this study, highlighted with a star. The scale bar represents the number of nucleotide substitutions per site.</p>
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22 pages, 762 KiB  
Article
Bayesian Estimation of the True Bovine Brucellosis Prevalence in Vaccinated and Non-Vaccinated Ecuadorian Cattle Populations, and the Sensitivity and Specificity of a Competitive and Indirect ELISA Using a New Synthetic Antigen
by Ana Dolores Garrido Haro, Margoth Yolanda Barrionuevo Samaniego, Paola Moreno-Caballeros, Alexandra Burbano-Enríquez, Verónica Alexandra Salas Torres, María Cristina Galante Mulki, Constance Wielick, Jorge Ron-Román and Claude Saegerman
Microorganisms 2025, 13(1), 69; https://doi.org/10.3390/microorganisms13010069 - 2 Jan 2025
Viewed by 309
Abstract
Bovine brucellosis (bB) is a zoonosis mainly caused by the Brucella abortus species in cattle. Bovine brucellosis can present with either a range of clinical symptoms, including spontaneous abortions in the last trimester of pregnancy, retained fetal membranes, and decreased milk production, or [...] Read more.
Bovine brucellosis (bB) is a zoonosis mainly caused by the Brucella abortus species in cattle. Bovine brucellosis can present with either a range of clinical symptoms, including spontaneous abortions in the last trimester of pregnancy, retained fetal membranes, and decreased milk production, or it can be asymptomatic. In Ecuador, vaccination against bB with S19 and/or RB51 is not mandatory and is the responsibility of the farmer. As serology is a convenient method for detecting antibodies against Brucella, evaluating the diagnostic performance and discriminative ability of such tests in various epidemiological settings is required. To estimate and compare the diagnostic sensitivity (Se) and specificity (Sp) of two screening tests, a new competitive (cELISA) and an indirect ELISA based on a new synthetic antigen (iELISA), a randomized, stratified, cross-sectional, serological survey was performed on the cattle population (3299 bovine sera from 223 farms) in continental Ecuador. A Bayesian approach was used to evaluate the two tests by estimating their respective diagnostic Se and Sp, as well as the true prevalence of bB in different sub-populations (non-vaccinated, vaccinated with S19 or RB51). The Se of both tests was similar across Bayesian models, with values around 94%. In contrast, the Sp of the iELISA, ranging between 97 and 98%, was significantly higher than that of the cELISA, which was approximately 94–95%. The true prevalence of bB was 1.63% (95% CrI: 0.56–2.54) in non-vaccinated cattle, decreased to 0.97% (95% CrI: 0.005–2.54) in S19-vaccinated cattle and was 2.75% (95% CrI: 0.50–5.32) in RB51-vaccinated cattle. The results of this study suggest that, with similar Se and higher Sp, the iELISA based on an innovative synthetic antigen (which is more standardizable) should be recommended as a possible screening test for bB in Ecuador. Also, the proposed approach suggests insights into the quality of the vaccination campaign and highlights the need for refining the Ecuadorian national brucellosis control program. Full article
(This article belongs to the Special Issue Epidemiology and Control Strategies for Brucellosis)
16 pages, 2354 KiB  
Review
Why Olive Produces Many More Flowers than Fruit—A Critical Analysis
by Julián Cuevas
Horticulturae 2025, 11(1), 26; https://doi.org/10.3390/horticulturae11010026 - 2 Jan 2025
Viewed by 220
Abstract
Olive (Olea europaea L.) trees produce many more flowers than fruit. In an “on” year, an adult olive tree may produce as many as 500,000 flowers, but 98% of them will drop soon after bloom as unfertilized flowers or juvenile fruit. This [...] Read more.
Olive (Olea europaea L.) trees produce many more flowers than fruit. In an “on” year, an adult olive tree may produce as many as 500,000 flowers, but 98% of them will drop soon after bloom as unfertilized flowers or juvenile fruit. This waste of resources that could be better invested in fruit reaching maturation requires an explanation. Several, not mutually exclusive, hypotheses explaining the possible significance of heavy flowering followed by massive and premature flower and fruit abscission are analyzed and compared based on previously published works and recent observations on olive reproductive biology. The results suggest that olive trees selectively abort fruits to enhance the quality of the seeds in the surviving fruits. Additionally, a considerable proportion of flowers appears to contribute to the male fitness of the plant by increasing pollen export. Conversely, the hypotheses attributing to resource limitation, pollination deficits, pollinator attraction, or extra flowers functioning as an ovary reserve, must be rejected for explaining the ultimate functions of massive flower production. Implications for olive orchard management are discussed. Full article
(This article belongs to the Special Issue Advances in Developmental Biology in Tree Fruit and Nut Crops)
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<p>Massive bloom in an olive tree during its “on” year. Panicles develop clustered on 1-year-old shoots. Source: own author.</p>
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<p>Olive panicles showing hermaphrodite and staminate flowers. (<b>Left</b>): Intact panicle. (<b>Right</b>): petals removed to better expose the well-developed pistil (white arrow) or the rudimentary pistil (black arrow). Source: Seifi et al. [<a href="#B8-horticulturae-11-00026" class="html-bibr">8</a>]; with permission.</p>
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<p>Olive pollen clouds commonly seen during blooming season in Andalusia (Spain). Source: José Angel García, Ideal 6 May 2020.</p>
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<p>Fruitful shoots from an “on” and an “off” olive tree. Despite “off” trees setting a proportionally higher number of flowers, the total number of fruits is still much larger in “on” trees due to the much higher number of inflorescences. Note the different shoot lengths. Fruit ripening occurs earlier in “off” trees. Source: own author.</p>
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<p>Scanning electron microscopy images of hermaphrodite (<b>left</b>) and staminate (<b>right</b>) olive flowers. (<b>A</b>). Detail of a well-developed stigma (SG) adjacent to one stamen (ST). (<b>B</b>). Pistil-aborted flower showing a rudimentary pistil (PI). (<b>C</b>). Pollen germination on fully developed stigma papillae. (<b>D</b>). Functionally male, late pistil-aborted flower displaying stigma with non-functional papillae. Source: own author [<a href="#B42-horticulturae-11-00026" class="html-bibr">42</a>].</p>
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<p>Composition of pollen tube attrition in cross-pollinated olive flowers. (<b>Left</b>): The flower has been dissected, softened with NaOH 1N, squashed, and stained with aniline blue, then observed under fluorescence microscopy. (<b>Right</b>): Different pistil sections showing progressive pollen tube attrition. (<b>Up</b>): stigma; (<b>down</b>): style. Red arrows indicate the “winner” pollen tube. Source: own author.</p>
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10 pages, 546 KiB  
Article
Outbreak of Salmonella enterica subsp. enterica Serovar Napoli on a Dairy Cow Farm
by Matteo Ricchi, Anita Filippi, Erika Scaltriti, Martina Tambassi, Stefano Pongolini, Luca Bolzoni, Alice Prosperi, Camilla Torreggiani, Medardo Cammi, Alessandro Chiatante, Norma Arrigoni, Elisa Massella, Andrea Luppi and Chiara Garbarino
Animals 2025, 15(1), 79; https://doi.org/10.3390/ani15010079 - 2 Jan 2025
Viewed by 401
Abstract
Salmonella is diffused worldwide, and Salmonella enterica subsp. enterica is spread worldwide with many serovars associated with the infection of domestic bovines. The most spread are S. Dublin, S. Typhimurium and S. Infantis. S. Napoli is, however very rarely reported in [...] Read more.
Salmonella is diffused worldwide, and Salmonella enterica subsp. enterica is spread worldwide with many serovars associated with the infection of domestic bovines. The most spread are S. Dublin, S. Typhimurium and S. Infantis. S. Napoli is, however very rarely reported in domestic ruminants. Here, we report an outbreak of S. Napoli on a dairy cow farm in Northern Italy (Piacenza). A total of 18 S. Napoli isolates were recovered from aborted fetuses, feces, tissues and environmental samples. Whole genome sequencing suggested that all isolates belonged to the same cluster. After the application of stringent biocontainment and biosecurity measures, no further cases were reported. However, four months after the first case, the serovar was still isolated in environmental samples, underlying the importance of adopting the correct biosecurity and biocontainment measures in order to prevent the circulation and transmission of Salmonella within the farm. Full article
(This article belongs to the Section Animal Welfare)
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<p>Phylogenetic tree showing all <span class="html-italic">S</span>. Napoli isolates recovered in the farm and four other human strains from the same region and year of isolation used as outgroup. In red: animal samples and in green: environmental samples. Bootstrap values &lt;59 are reported at the nodes.</p>
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11 pages, 766 KiB  
Article
Adverse Pregnancy Outcomes in Patients with Polycystic Ovary Syndrome with Pre-Conceptional Hyperandrogenism: A Multi-Institutional Registry-Based Retrospective Cohort Study
by Yi-Ting Chang, Ming-Jer Chen, Wei-Szu Lin, Ching-Heng Lin and Jui-Chun Chang
J. Clin. Med. 2025, 14(1), 123; https://doi.org/10.3390/jcm14010123 - 28 Dec 2024
Viewed by 307
Abstract
Background/Objectives: Women with polycystic ovarian syndrome (PCOS) are at higher risk for pregnancy complications. The PCOS population is heterogeneous, with different phenotypes linked to varying risks of adverse outcomes. However, literature on pre-conceptional hyperandrogenism is limited and based on small sample sizes. [...] Read more.
Background/Objectives: Women with polycystic ovarian syndrome (PCOS) are at higher risk for pregnancy complications. The PCOS population is heterogeneous, with different phenotypes linked to varying risks of adverse outcomes. However, literature on pre-conceptional hyperandrogenism is limited and based on small sample sizes. Methods: This multi-institutional registry-based retrospective cohort study included pregnant patients diagnosed with PCOS with or without pre-conceptional hyperandrogenism. Utilizing the TriNetX platform, one-to-one propensity score matching was conducted to adjust for confounding factors. Exclusion criteria included multiple pregnancies and patients who received assisted reproductive technology, oral contraceptives, or spironolactone. 571 patients with PCOS and pre-conceptional hyperandrogenism and 13,465 patients with PCOS without hyperandrogenism were identified. Post-propensity matching, each cohort consisted of 564 patients. Results: Pregnant women diagnosed with PCOS and pre-conceptional hyperandrogenism showed a higher risk of large for gestational age (6.6% vs. 3.9%, OR = 1.73, 95% CI [1.007–2.972], p-value = 0.045) and preterm birth (10.3% vs. 5.9%, OR = 1.844, 95% CI [1.183–2.876], p-value = 0.006), but had no significant increase in the risk of gestational hypertension, preeclampsia/eclampsia, gestational diabetes, missed abortion, intrauterine growth restriction, placenta abruption, or cesarean section. Conclusions: Women with PCOS and pre-conceptional hyperandrogenism have an increased risk of pregnancy complications, especially large for gestational age and preterm birth. Further research is needed to clarify the underlying mechanisms, and whether treatment can improve outcomes. Full article
(This article belongs to the Section Reproductive Medicine & Andrology)
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<p>Patient enrollment algorithm. Abbreviations: HCOs, healthcare organizations; PCOS, polycystic ovarian syndrome; COCs, combined oral contraceptives; BMI, body mass index.</p>
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5 pages, 141 KiB  
Editorial
Genetic and Epigenetic Insights into Pregnancy-Related Complications
by Nihar R. Nayak, Akhilesh Srivastava, Manoj Kumar Jena, Anthony Odibo and Gary Sutkin
Genes 2025, 16(1), 1; https://doi.org/10.3390/genes16010001 - 24 Dec 2024
Viewed by 441
Abstract
Placental dysfunction is a leading cause of numerous pregnancy complications, including preeclampsia, preterm birth, fetal growth restrictions, placental abruption, and late spontaneous abortion [...] Full article
23 pages, 16678 KiB  
Article
An Epidemiological Study on Salmonella in Tibetan Yaks from the Qinghai–Tibet Plateau Area in China
by Dengyu Li, Kaiqin Zhang, Xiaofeng Xue, Zhanchun Bai, La Yang, Jingjing Qi and Sizhu Suolang
Animals 2024, 14(24), 3697; https://doi.org/10.3390/ani14243697 - 21 Dec 2024
Viewed by 609
Abstract
Salmonella is an important foodborne pathogen that can cause a range of illnesses in humans; it has also been a key focus for monitoring in the field of public health, including gastroenteritis, sepsis, and arthritis, and can also cause a decline in egg [...] Read more.
Salmonella is an important foodborne pathogen that can cause a range of illnesses in humans; it has also been a key focus for monitoring in the field of public health, including gastroenteritis, sepsis, and arthritis, and can also cause a decline in egg production in poultry and diarrhea and abortion in livestock, leading to death in severe cases, resulting in huge economic losses. This study aimed to investigate the isolation rate, antimicrobial resistance, serotypes, and genetic diversity of Salmonella isolated from yak feces in various regions on the Qinghai–Tibet Plateau. A total of 1222 samples of yak dung were collected from major cities in the Qinghai–Tibet Plateau area, and the sensitivity of the isolated bacteria to 10 major classes of antibiotics was determined using the K-B paper disk diffusion method for drug susceptibility. Meanwhile, the serotypes of the isolated bacteria were analyzed using the plate agglutination test for serum antigens, and their carriage of drug resistance and virulence genes was determined using PCR and gel electrophoresis experiments. The isolated bacteria were also classified using MLST (Multi-Locus Sequence Typing). The overall isolation rate for Salmonella was 18.25% (223/1222), and the results of the antibiotic susceptibility tests showed that 98.65% (220/223) of the isolated bacteria were resistant to multiple antibiotics. In the 223 isolates of Salmonella, eight classes of 20 different resistance genes, 30 serotypes, and 15 different types of virulence genes were detected. The MLST analysis identified 45 distinct sequence types (STs), including five clonal complexes, of which ST34, ST11, and ST19 were the most common. These findings contribute valuable information about strain resources, genetic profiles, and typing data for Salmonella in the Qinghai–Tibet Plateau area, facilitating improved bacterial surveillance, identification, and control in yak populations. They also provide certain data supplements for animal Salmonella infections globally, filling research gaps. Full article
(This article belongs to the Section Cattle)
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<p>Geographical distribution of sample collection sites (marked with ▲).</p>
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<p>Antimicrobial resistance heatmap of 223 <span class="html-italic">Salmonella</span> isolates from the Tibetan Plateau. The regions from which samples were sourced are on the left vertical axis, and the antimicrobials (1–26) are on the horizontal axis. The color scale bar represents the diameter of the inhibition zone, ranging from 0 to 40 mm. (1) Cefotaxime; (2) cefazolin; (3) ampicillin; (4) cefuroxime; (5) cefoperazone; (6) penicillin; (7) cephalexin; (8) oxacillin; (9) azithromycin; (10) erythromycin; (11) gentamicin; (12) kanamycin; (13) streptomycin; (14) minocycline; (15) doxycycline; (16) tetracycline; (17) lincomycin; (18) chloramphenicol; (19) florfenicol; (20) vancomycin; (21) bacitracin; (22) polymyxin B; (23) ciprofloxacin; (24) nalidixic acid; (25) furazolidone; and (26) sulfamethoxazole.</p>
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<p>Specific antimicrobial resistance of <span class="html-italic">Salmonella</span> on the Qinghai–Tibet Plateau. (<b>A</b>) Multidrug resistance of 223 strains of <span class="html-italic">Salmonella</span> from yaks on the Qinghai–Tibet Plateau. (<b>B</b>) The specific number of drug-resistant <span class="html-italic">Salmonella</span> isolates from yaks on the Qinghai–Tibet Plateau. (<b>C</b>) The antimicrobial resistance differences in <span class="html-italic">Salmonella</span> isolates from Tibet and Qinghai. The bar chart shows the percentage of resistant strains. * indicates a significant difference (<span class="html-italic">p</span> &lt; 0.05). ** indicates a highly significant difference (<span class="html-italic">p</span> &lt; 0.01).</p>
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<p>Prevalence of <span class="html-italic">Salmonella</span> resistance genes on the Qinghai–Tibet Plateau. (<b>A</b>) Antimicrobial resistance gene carriage in 223 <span class="html-italic">Salmonella</span> isolates from yaks from the Qinghai–Tibet Plateau region. (<b>B</b>) Differences in the carriage of antimicrobial resistance genes in <span class="html-italic">Salmonella</span> isolates from Tibet and Qinghai. * indicates a significant difference (<span class="html-italic">p</span> &lt; 0.05). ** indicates a highly significant difference (<span class="html-italic">p</span> &lt; 0.01).</p>
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<p>Relationships between <span class="html-italic">Salmonella</span> pathogenicity genes in the Qinghai–Tibet Plateau. (<b>A</b>) The presence of virulence genes in 223 strains of <span class="html-italic">Salmonella</span> isolated from yaks in the Qinghai–Tibet Plateau region. (<b>B</b>) Differences in the presence of virulence genes in <span class="html-italic">Salmonella</span> isolates from Tibet and Qinghai. * indicates a significant difference (<span class="html-italic">p</span> &lt; 0.05). ** indicates a highly significant difference (<span class="html-italic">p</span> &lt; 0.01).</p>
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<p>Information on serotypes of <span class="html-italic">Salmonella</span> on the Qinghai–Tibet Plateau. (<b>A</b>) The distribution of the serotypes of 223 strains of <span class="html-italic">Salmonella</span> isolated from yaks in the Qinghai–Tibet Plateau region. (<b>B</b>) The distribution of the serotypes of <span class="html-italic">Salmonella</span> isolates in different regions of Tibet and Qinghai.</p>
Full article ">Figure 6 Cont.
<p>Information on serotypes of <span class="html-italic">Salmonella</span> on the Qinghai–Tibet Plateau. (<b>A</b>) The distribution of the serotypes of 223 strains of <span class="html-italic">Salmonella</span> isolated from yaks in the Qinghai–Tibet Plateau region. (<b>B</b>) The distribution of the serotypes of <span class="html-italic">Salmonella</span> isolates in different regions of Tibet and Qinghai.</p>
Full article ">Figure 6 Cont.
<p>Information on serotypes of <span class="html-italic">Salmonella</span> on the Qinghai–Tibet Plateau. (<b>A</b>) The distribution of the serotypes of 223 strains of <span class="html-italic">Salmonella</span> isolated from yaks in the Qinghai–Tibet Plateau region. (<b>B</b>) The distribution of the serotypes of <span class="html-italic">Salmonella</span> isolates in different regions of Tibet and Qinghai.</p>
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<p>Minimum spanning tree for <span class="html-italic">Salmonella</span> on the Qinghai–Tibet Plateau. (<b>A</b>) Minimum evolution tree generated from regional characteristics. (<b>B</b>) Minimum evolution tree generated from serotypes as features.</p>
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<p>Minimum spanning tree for <span class="html-italic">Salmonella</span> on the Qinghai–Tibet Plateau. (<b>A</b>) Minimum evolution tree generated from regional characteristics. (<b>B</b>) Minimum evolution tree generated from serotypes as features.</p>
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<p>Phylogenetic tree and allelic profile of 223 strains of <span class="html-italic">Salmonella</span> isolated from yaks, along with serotype information.</p>
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16 pages, 2500 KiB  
Article
Computer-Aided Diagnosis in Spontaneous Abortion: A Histopathology Dataset and Benchmark for Products of Conception
by Tahir Mahmood, Zeeshan Ullah, Atif Latif, Binish Arif Sultan, Muhammad Zubair, Zahid Ullah, AbuZar Ansari, Talat Zehra, Shahzad Ahmed and Naqqash Dilshad
Diagnostics 2024, 14(24), 2877; https://doi.org/10.3390/diagnostics14242877 - 21 Dec 2024
Viewed by 459
Abstract
Spontaneous abortion, commonly known as miscarriage, is a significant concern during early pregnancy. Histopathological examination of tissue samples is a widely used method to diagnose and classify tissue phenotypes found in products of conception (POC) after spontaneous abortion. Background: Histopathological examination is subjective [...] Read more.
Spontaneous abortion, commonly known as miscarriage, is a significant concern during early pregnancy. Histopathological examination of tissue samples is a widely used method to diagnose and classify tissue phenotypes found in products of conception (POC) after spontaneous abortion. Background: Histopathological examination is subjective and dependent on the skill and experience of the examiner. In recent years, artificial intelligence (AI)-based techniques have emerged as a promising tool in medical imaging, offering the potential to revolutionize tissue phenotyping and improve the accuracy and reliability of the histopathological examination process. The goal of this study was to investigate the use of AI techniques for the detection of various tissue phenotypes in POC after spontaneous abortion and evaluate the accuracy and reliability of these techniques compared to traditional manual methods. Methods: We present a novel publicly available dataset named HistoPoC, which is believed to be the first of its kind, focusing on spontaneous abortion (miscarriage) in early pregnancy. A diverse dataset of 5666 annotated images was prepared from previously diagnosed cases of POC from Atia General Hospital, Karachi, Pakistan, for this purpose. The digital images were prepared at 10× through a camera-connected microscope by a consultant histopathologist. Results: The dataset’s effectiveness was validated using several deep learning-based models, demonstrating its applicability and supporting its use in intelligent diagnostic systems. Conclusions: The insights gained from this study could illuminate the causes of spontaneous abortion and guide the development of novel treatments. Additionally, this study could contribute to advancements in the field of tissue phenotyping and the wider application of deep learning techniques in medical diagnostics and treatment. Full article
(This article belongs to the Special Issue Latest News in Digital Pathology)
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<p>Illustration of the generic framework employed in this study.</p>
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<p>Example images from the dataset captured under 10× magnification using a camera-connected microscope. (<b>a</b>) Chorionic villi, (<b>b</b>) decidual tissue, (<b>c</b>) hemorrhage, and (<b>d</b>) trophoblastic tissue.</p>
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<p>Example of chorionic villi tissue from the dataset captured under 10× magnification using a camera-connected microscope.</p>
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<p>Example of decidual tissue from the dataset captured under 10× magnification using a camera-connected microscope.</p>
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<p>Example of hemorrhage tissue from the dataset captured under 10× magnification using a camera-connected microscope.</p>
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<p>Example of trophoblastic tissue from the dataset captured under 10× magnification using a camera-connected microscope.</p>
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13 pages, 783 KiB  
Article
Molecular Testing of Zoonotic Bacteria in Cattle, Sheep, and Goat Abortion Cases in Botswana
by Boitumelo M. Modise-Tlotleng, Sununguko W. Mpoloka, Tirumala B. K. Settypalli, Joseph Hyera, Tebogo Kgotlele, Kago Kumile, Mosarwa E. Sechele, Obuile O. Raboloko, Chandapiwa Marobela-Raborokgwe, Gerrit J. Viljoen, Giovanni Cattoli and Charles E. Lamien
Microorganisms 2024, 12(12), 2644; https://doi.org/10.3390/microorganisms12122644 - 20 Dec 2024
Viewed by 585
Abstract
Abortion is one of the major causes of economic losses in livestock production worldwide. Because several factors can lead to abortion in cattle, sheep and goats, laboratory diagnosis, including the molecular detection of pathogens causing abortion, is often necessary. Bacterial zoonotic diseases such [...] Read more.
Abortion is one of the major causes of economic losses in livestock production worldwide. Because several factors can lead to abortion in cattle, sheep and goats, laboratory diagnosis, including the molecular detection of pathogens causing abortion, is often necessary. Bacterial zoonotic diseases such as brucellosis, coxiellosis, leptospirosis, and listeriosis have been implicated in livestock abortion, but they are under diagnosed and under-reported in most developing countries, including Botswana. This study applied a recently developed multiplex high-resolution melting analysis technique, coupled with singleplex qPCR assays, to investigate abortions in livestock in Botswana, using 152 samples from cattle, sheep, and goat abortion cases. Brucella spp. were the most frequent pathogen detected, with an overall frequency of 21.1%, followed by Coxiella burnetii with 19.1%. Listeria monocytogenes and Leptospira spp. were not detected in any of specimens samples investigated. Mixed infections with Brucella spp. and C. burnetii were observed in 35% specimes examined. There was a good agreement between the multiplex qPCR-HRM and singleplex qPCR for detecting Brucella spp. and C. burnetii. This study is the first report on the syndromic testing of abortion-causing pathogens in Botswana. It shows the importance of molecular methods in the differential diagnosis of abortion-causing diseases in domestic ruminants. Full article
(This article belongs to the Section Veterinary Microbiology)
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<p>Distribution of <span class="html-italic">Brucella-</span> and <span class="html-italic">Coxiella</span>-positive cases from 2014 to 2021.</p>
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<p>Map of Botswana showing distribution of pathogens per location.</p>
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10 pages, 296 KiB  
Article
Epidemiology of Bovine Neosporosis in Relation to Socioeconomic, Demographic, and Transmissibility Factors in Dual-Purpose Production Systems in Colombia
by Cesar A. Murcia-Mono, Sergio Falla-Tapias, Britney K. Cabrera-Ospina, Jahir O. Vargas-Domínguez and William O. Burgos-Paz
Epidemiologia 2024, 5(4), 828-837; https://doi.org/10.3390/epidemiologia5040056 - 19 Dec 2024
Viewed by 740
Abstract
Introduction: Bovine neosporosis represents a significant threat to reproduction and production in livestock systems worldwide. This disease is caused by the protozoan Neospora caninum, resulting in abortions of cows and neurological signs in newborn calves. This leads to significant economic losses, decreasing [...] Read more.
Introduction: Bovine neosporosis represents a significant threat to reproduction and production in livestock systems worldwide. This disease is caused by the protozoan Neospora caninum, resulting in abortions of cows and neurological signs in newborn calves. This leads to significant economic losses, decreasing meat and milk production, especially in tropical regions. The infection has an endogenous and exogenous cycle of transmission involving dogs that shed the oocysts, with the highest transmission successes in humid areas. Similarly, there is a lack of knowledge about the epidemiological risk factors and management practices involved in the transmission success in tropical humid regions. Methods: In this sense, a cross-sectional epidemiological survey was conducted on 150 farms from 24 municipalities of the Huila area. A total of 360 cattle were sampled, and information about the production system was collected using a structured poll with 128 questions. Results: In these cattle, 53% (191/360) were positive for antibodies against Neospora caninum using ELISA. The logistic regression analysis using the information collected from the poll identified the presence of flooring type, water access, production systems, and feed management as risk factors. Among the protective factors were identified the geographical area, molasses supplementation, and biosecurity practices such as animal separation and access control. Discussion: This study identified for the first time the epidemiological risk factors associated mainly with the exogenous cycle of neosporosis. The present study contributes to the design of intervention strategies oriented to minimize the impact of parasitism in Colombian herds. Full article
18 pages, 3901 KiB  
Article
A Transcriptome Approach Evaluating the Effects of Atractylenolide I on the Secretion of Estradiol and Progesterone in Feline Ovarian Granulosa Cells
by Yuli Guo, Junping Liu, Shuangyi Zhang, Di Sun, Zhiying Dong and Jinshan Cao
Vet. Sci. 2024, 11(12), 663; https://doi.org/10.3390/vetsci11120663 - 17 Dec 2024
Viewed by 648
Abstract
Atractylodes macrocephala Koidz (AMK) as an oriental medicine has been used in the treatment of threatened abortion. Atractylenolide I (AT-I) is one of the major bioactive components of AMK. This study aimed to investigate the effect of AT-I on the secretion of estradiol [...] Read more.
Atractylodes macrocephala Koidz (AMK) as an oriental medicine has been used in the treatment of threatened abortion. Atractylenolide I (AT-I) is one of the major bioactive components of AMK. This study aimed to investigate the effect of AT-I on the secretion of estradiol (E2) and progesterone (P4) in feline ovarian granulosa cells (FOGCs), which is necessary for pregnancy. At first, the proliferation of FOGCs after AT-I treatment was measured by CCK-8. Then, the synthesis of E2 and P4 were measured by ELISA. Lastly, transcriptome sequencing was used to detect the DEGs in the FOGCs, and RNA-seq results were verified by RT-qPCR and biochemical verification. It was found that AT-I could promote proliferation and the secretion of E2 and P4 in FOGCs; after AT-I treatment, 137 significant DEGs were observed, out of which 49 were up-regulated and 88 down-regulated. The DEGs revealed significant enrichment of 52 GO terms throughout the differentiation process (p < 0.05), as deciphered by Gene Ontology enrichment analysis. Kyoto Encyclopedia of Genes and Genomes analysis manifested that the DEGs were successfully annotated as members of 155 pathways, with 23 significantly enriched (p < 0.05). A relatively high number of genes were enriched for the cholesterol metabolism, ovarian steroidogenesis, and biosynthesis of unsaturated fatty acids. Furthermore, the contents of the total cholesterol and low-density lipoprotein cholesterol were decreased by AT-I treatment in the cell culture supernatant. The results indicated that AT-I could increase the ability of FOGCs to secrete E2 and P4, which might be achieved by activation of cholesterol metabolism. Full article
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<p>Chemical structure of Atractylenolide-I (AT-I).</p>
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<p>Morphological observation and growth curve of the primary FOGCs. (<b>A</b>) The primary FOGCs attach to the substrate around the oocytes and begin to display an elongated or fibroblastic property within the first 24 h of culture. (<b>B</b>) Growth curve of cultivated cells within 48 h. (<b>C</b>) The HE staining of cultivated cells on the substrate. (<b>D</b>) The Giemsa staining of cultivated cells on the substrate. (<b>E</b>) Immunofluorescence of cultivated cells on the substrate: FSHR (green) as the marker expressed in FOGCs; negative-control group and nuclei with DAPI staining (blue). Values are expressed as mean ± S.D. (n = 4). *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Effect of AT-I on the FOGCs. (<b>A</b>) Treatment with AT-I in 1, 3, 10, and 30 µmol/L could effectively promote FOGC proliferation from 12 h to 48 h; 100 µmol/L significantly increased FOGC proliferation after 36 h and the increased effect of 300 µmol/L happened at 48 h (<span class="html-italic">p</span> &lt; 0.05). (<b>B</b>) After 10 μmol/L was AT-I-treated for 36 h, the density of granulosa cells increased, and cell viability increased significantly. Values are expressed as mean ± S.D. (n = 4). *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01; ns, not statistically significant.</p>
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<p>The effect of 10 μmol/L AT-I on the synthesis of E<sub>2</sub> and P<sub>4</sub> in FOGCs. (<b>A</b>) AT-I could significantly promote the E<sub>2</sub> secretion in FOGCs at 24 h and 36 h, while the increased tendency weakened at 48 h. (<b>B</b>) The P<sub>4</sub> concentration was significantly increased by AT-I treatment from 12 h to 48 h. (<b>C</b>) The ratio of P<sub>4</sub>/E<sub>2</sub> indicated that the E<sub>2</sub> secretion was more obviously increased than P<sub>4</sub> secretion in FOGCs within 36 h after AT-I treatment. However, the secretion pattern of E<sub>2</sub> and P<sub>4</sub> changed at 48 h, in which the FOGCs secreted P<sub>4</sub> more obviously than they secreted E<sub>2</sub>. Values are expressed as mean ± S.D. (n = 3). **, <span class="html-italic">p</span> &lt; 0.01; ns, not statistically significant.</p>
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<p>Volcano graph of 137 differentially expressed genes between two groups. Red represents up-regulated DEGs; blue represents down-regulated DEGs.</p>
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<p>GO classifications of genes in FOGCs. The abscissa is the GO classification, and the ordinate is the number of genes in each category.</p>
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<p>KEGG pathway classification of genes in FOGCs. The abscissa is the name of the KEGG metabolic pathway, and the ordinate is the ratio of the number of genes annotated to the pathway and the number of genes in the annotated genes.</p>
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<p>The significantly enriched KEGG pathway of DEGs. Rich factor, number of differentially expressed genes/total number of genes in this KEGG pathway. The larger the value, the greater the enrichment.</p>
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<p>Verification of transcriptome results after 10 µmol/L AT-I treatment for 36 h. (<b>A</b>) Relative quantification of DEGs by RT-qPCR. The expression of each gene was normalized to the average expression of the endogenous reference gene GAPDH. (<b>B</b>) Biochemical verification. After AT-I treatment, the contents of total cholesterol and LDL-cholesterol both declined. The values are expressed as mean ± S.D. (n = 3). *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt;0.01; ns, not statistically significant.</p>
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20 pages, 9291 KiB  
Article
Development Using Bioluminescence Imaging of a Recombinant Anguillid Herpesvirus 1 Vaccine Candidate Associated with Normal Replication In Vitro but Abortive Infection In Vivo
by Haiyan Zhang, Arun Sridhar, Natacha Delrez, Bo He, Sophie Fourny, Yuan Gao, Owen Donohoe and Alain F. C. Vanderplasschen
Vaccines 2024, 12(12), 1423; https://doi.org/10.3390/vaccines12121423 - 17 Dec 2024
Viewed by 653
Abstract
Background/Objectives: Anguillid herpesvirus 1 (AngHV-1) (recently renamed Cyvirus anguillidallo 1) is the etiologic agent of a lethal disease that affects several eel species. It is thought to be one of the main infectious agents causing a population decline in wild eels and economic [...] Read more.
Background/Objectives: Anguillid herpesvirus 1 (AngHV-1) (recently renamed Cyvirus anguillidallo 1) is the etiologic agent of a lethal disease that affects several eel species. It is thought to be one of the main infectious agents causing a population decline in wild eels and economic loss within the eel aquaculture sector. To date, no vaccines are available against AngHV-1. Recently, we developed a safe and efficacious live attenuated recombinant vaccine against Cyprinid herpesvirus 3 (CyHV-3). This CyHV-3 recombinant vaccine encodes a deletion of ORF57. Orthologues of CyHV-3 ORF57 exist in Cyprinid herpesvirus 2 (CyHV-2, ORF57) and AngHV-1 (ORF35). Methods: In the present study, using recombinant strains and bioluminescent in vivo imaging, we investigated the effect of AngHV-1 ORF35 deletion on virus replication in vitro, virulence in vivo, and the potential of an AngHV-1 ORF35-deleted recombinant as a vaccine candidate for the mass vaccination of eels by immersion. With this goal in mind, we produced ORF35-deleted recombinants using two parental strains: a UK strain and a recombinant derived from the former strain by insertion of a Luciferase–GFP reporter cassette into a non-coding intergenic region. Results: Analyses of ORF35-deleted recombinants led to the following observations: (i) AngHV-1 ORF35 is not essential for viral growth in cell culture, and its deletion does not affect the production of extracellular virions despite reducing the size of viral plaque. (ii) In contrast to what has been observed for CyHV-3 ORF57 and CyHV-2 ORF57, in vivo bioluminescent analyses revealed that AngHV-1 ORF35 is an essential virulence factor and that its deletion led to abortive infection in vivo. (iii) Inoculation of the AngHV-1 ORF35-deleted recombinant by immersion induced a protective immune response against a wild-type challenge. This protection was shown to be dose-dependent and to rely on the infectivity of AngHV-1 ORF35-deleted virions. Conclusions: This study suggests that the AngHV-1 ORF35 protein has singular properties compared to its orthologues encoded by CyHV-2 and CyHV-3. It also supports the potential of AngHV-1 ORF35-deleted recombinants for the mass vaccination of eels by immersion. Full article
(This article belongs to the Special Issue Animal Herpesviruses)
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<p>Schematic representation of the strategy used to produce AngHV-1 recombinants by homologous directed recombination (HDR). (<b>A</b>) Flowchart of the production of the UK Luc and ORF35 Del recombinant strains by HDR in eukaryotic cells. (<b>B</b>) Genotype of the UK parental strain and derived recombinant strains for the ORF32–ORF33 intergenic region and the ORF35 locus. WT, wild-type; Luc, inserted LucGFP cassette; Del, deleted. (<b>C</b>) A schematic representation of the genome structure of UK Luc. The genome of AngHV-1 flanked by two terminal repeats (LTR and RTR) and the intergenic ORF32–ORF33 genome region are shown at the top. (<b>D</b>) Schematic representation of the genome structure of the UK ORF35 Del recombinant. The genome of AngHV-1 flanked by two terminal repeats (LTR and RTR) and the ORF35 genome region are shown at the top. In panels (<b>C</b>,<b>D</b>), SacI restriction sites and predicted restriction fragments (in kb) are shown. Coordinates are those of the AngHV-1 reference strain available in GenBank (accession number: MW580855.1).</p>
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<p>Characterization of AngHV-1 recombinant strains. (<b>A</b>) Transcriptional analysis of genes ORF32, ORF33, ORF34, ORF35, ORF36, and ORF55 expressed by the indicated strains of AngHV-1. ORF55 (AngHV-1 DNA polymerase) expression was used as a control. Marker sizes (MSs) in base pairs (bps) are indicated on the left. The left part and the right part of the figure represent the results of the PCR performed on cDNA and RNA, respectively. (<b>B</b>) Expression of reporter genes. EK-1 cells grown in 12-well plates were infected with the indicated strains, then overlaid with a medium containing CMC. At 4 dpi, infected cells were analyzed for the expression of bioluminescence and epifluorescence. The Luc signal was detected using the IVIS system (left frame). The reporters (copGFP and mCherry) and immunofluorescent staining (anti-AngHV-1) were detected by epifluorescent microscopy. Plaques of UK and UK ORF35 Del were revealed by indirect immunofluorescent staining (anti-AngHV-1) (right frame). (<b>C</b>) The replication kinetics and (<b>D</b>) viral plaque sizes of the ORF35-deleted strains were compared with those of the parental UK and UK Luc strains. (<b>E</b>) Luc expression of AngHV-1 recombinant strains (UK Luc and UK Luc ORF35 Del). The replication kinetic data represent the mean ± SEM of triplicate measurements. The data on the plaque area are the mean ± SEM of twenty measurements. The data on Luc expression are the mean ± SEM of triplicate measurements. The horizontal dashed line in panel <b>E</b> shows the mean + 3 SD of the data obtained for control non-infected cultures. Results of statistical comparisons between ORF35 Del strains and parental strains are indicated as follows: ns, no significant differences; *, <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of ORF35 deletion on AngHV-1 replication in vivo. (<b>A</b>) Flowchart of the experiment. At the time of inoculation, yellow eels (12.06 ± 2.72 g, mean ± SD) were mock-infected or infected with the indicated strains using different routes: IP injection of 200,000 pfu/eel or immersion in water containing 4000 pfu/mL or intradermal inoculation of 200,000 pfu/eel. At the indicated times post-infection, eels (<span class="html-italic">n</span> = 6, consisting of two eels from triplicate tanks) were imaged using an IVIS. (<b>B</b>) The effects of AngHV-1 infection routes: IP injection, immersion, and intradermal inoculation are presented in the left column, middle column, and right column, respectively. Average radiance (individual values, mean ± SEM) measured on the entire body surface of the fish, i.e., skin (individual values represent the mean values obtained for the left and right sides of each fish), gills (individual values represent the mean values obtained for the left and right gills), brain, heart, and gut-liver, were analyzed by IVIS (<span class="html-italic">n</span> = 6 per timepoint). The dashed line represents the threshold of positivity, which is the mean + 3 SD of the values obtained for the mock-infected fish (data not presented). The number of positive fish among the six analyzed fish is represented by bars (right axis). The average emitted radiances (<span class="html-italic">p</span>(<span class="html-italic">rad</span>)) of UK Luc were compared with UK Luc ORF35 Del using unpaired <span class="html-italic">t</span>-test (two-tail, Gaussian distribution) or Wilcoxon test (non-Gaussian distribution). The number of positive fish per group was compared between two strains using the Fisher–Pitman permutation test (<span class="html-italic">p</span>(<span class="html-italic">no</span>)). <span class="html-italic">p</span> values are represented as follows: ns, not significant; *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01; ***, <span class="html-italic">p</span> &lt; 0.001. (<b>C</b>) Representative images of IVIS data (skin, gills, and brain) are presented. Eels with the closest scores to the mean of each infection route were selected for image illustration.</p>
Full article ">Figure 3 Cont.
<p>Effect of ORF35 deletion on AngHV-1 replication in vivo. (<b>A</b>) Flowchart of the experiment. At the time of inoculation, yellow eels (12.06 ± 2.72 g, mean ± SD) were mock-infected or infected with the indicated strains using different routes: IP injection of 200,000 pfu/eel or immersion in water containing 4000 pfu/mL or intradermal inoculation of 200,000 pfu/eel. At the indicated times post-infection, eels (<span class="html-italic">n</span> = 6, consisting of two eels from triplicate tanks) were imaged using an IVIS. (<b>B</b>) The effects of AngHV-1 infection routes: IP injection, immersion, and intradermal inoculation are presented in the left column, middle column, and right column, respectively. Average radiance (individual values, mean ± SEM) measured on the entire body surface of the fish, i.e., skin (individual values represent the mean values obtained for the left and right sides of each fish), gills (individual values represent the mean values obtained for the left and right gills), brain, heart, and gut-liver, were analyzed by IVIS (<span class="html-italic">n</span> = 6 per timepoint). The dashed line represents the threshold of positivity, which is the mean + 3 SD of the values obtained for the mock-infected fish (data not presented). The number of positive fish among the six analyzed fish is represented by bars (right axis). The average emitted radiances (<span class="html-italic">p</span>(<span class="html-italic">rad</span>)) of UK Luc were compared with UK Luc ORF35 Del using unpaired <span class="html-italic">t</span>-test (two-tail, Gaussian distribution) or Wilcoxon test (non-Gaussian distribution). The number of positive fish per group was compared between two strains using the Fisher–Pitman permutation test (<span class="html-italic">p</span>(<span class="html-italic">no</span>)). <span class="html-italic">p</span> values are represented as follows: ns, not significant; *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01; ***, <span class="html-italic">p</span> &lt; 0.001. (<b>C</b>) Representative images of IVIS data (skin, gills, and brain) are presented. Eels with the closest scores to the mean of each infection route were selected for image illustration.</p>
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<p>Dose-protection effect conferred by UK ORF35 Del in vivo. (<b>A</b>) Flowchart of the experiment. At the time of primary inoculation, yellow eels (28.52 ± 7.30 g, mean ± SD) were mock-infected or infected for 2 h by immersion in water containing the indicated doses of UK ORF35 Del or 100,000 pfu/mL of UK ORF35 Del strain treated by psoralen/UV to inactivate virus infectivity. At 36 days post-primary inoculation, eels were infected for 2 h by immersion in water containing 4000 pfu/mL of UK Luc expressing luciferase as a reporter. At the indicated times post-secondary inoculation, eels (<span class="html-italic">n</span> = 6, consisting of two eels from triplicate tanks) were imaged using IVIS. (<b>B</b>) Average radiance (individual values, mean ± SEM) measured on the entire body surface of fish, i.e., skin (individual values represent the mean values obtained for the left and right sides of each fish), gills (individual values represent the mean values obtained for the left and right gills), brain, heart, and gut–liver, were analyzed by IVIS (<span class="html-italic">n</span> = 6 per timepoint). The average radiance (<span class="html-italic">p</span>(<span class="html-italic">rad</span>)) of each group was compared with the “primary mock-infected group” using a non-parametric Kruskal–Wallis test followed by multiple comparisons with the two-stage step-up method of Benjamini, Krieger, and Yekutieli. Throughout this panel, the data obtained for every individual eel (within each group) are represented by the same symbol to allow for a correlation of the data obtained for the different organs at a specific dpi. The dashed line represents the threshold of positivity, which was calculated by the mean + 3 SD of the values obtained for the mock fish. The number of positive fish among the six analyzed fish is represented by bars (right axis). The positive fish (<span class="html-italic">p</span>(<span class="html-italic">no</span>)) from each group was compared with the primary mock-infected group using the Fisher–Pitman permutation test. <span class="html-italic">p</span> values are represented as follows: ns, not significant; *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01. (<b>C</b>) Representative images of IVIS data (skin, gills, and brain) are presented in the lower panel. Eels with the closest scores to the mean of each group (mock; UK ORF35 Del, 100,000 pfu/mL; psoralen/UV-inactivated UK ORF35 Del, 100,000 pfu/mL; and mock-infected with UK Luc) were selected for image illustration.</p>
Full article ">Figure 4 Cont.
<p>Dose-protection effect conferred by UK ORF35 Del in vivo. (<b>A</b>) Flowchart of the experiment. At the time of primary inoculation, yellow eels (28.52 ± 7.30 g, mean ± SD) were mock-infected or infected for 2 h by immersion in water containing the indicated doses of UK ORF35 Del or 100,000 pfu/mL of UK ORF35 Del strain treated by psoralen/UV to inactivate virus infectivity. At 36 days post-primary inoculation, eels were infected for 2 h by immersion in water containing 4000 pfu/mL of UK Luc expressing luciferase as a reporter. At the indicated times post-secondary inoculation, eels (<span class="html-italic">n</span> = 6, consisting of two eels from triplicate tanks) were imaged using IVIS. (<b>B</b>) Average radiance (individual values, mean ± SEM) measured on the entire body surface of fish, i.e., skin (individual values represent the mean values obtained for the left and right sides of each fish), gills (individual values represent the mean values obtained for the left and right gills), brain, heart, and gut–liver, were analyzed by IVIS (<span class="html-italic">n</span> = 6 per timepoint). The average radiance (<span class="html-italic">p</span>(<span class="html-italic">rad</span>)) of each group was compared with the “primary mock-infected group” using a non-parametric Kruskal–Wallis test followed by multiple comparisons with the two-stage step-up method of Benjamini, Krieger, and Yekutieli. Throughout this panel, the data obtained for every individual eel (within each group) are represented by the same symbol to allow for a correlation of the data obtained for the different organs at a specific dpi. The dashed line represents the threshold of positivity, which was calculated by the mean + 3 SD of the values obtained for the mock fish. The number of positive fish among the six analyzed fish is represented by bars (right axis). The positive fish (<span class="html-italic">p</span>(<span class="html-italic">no</span>)) from each group was compared with the primary mock-infected group using the Fisher–Pitman permutation test. <span class="html-italic">p</span> values are represented as follows: ns, not significant; *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01. (<b>C</b>) Representative images of IVIS data (skin, gills, and brain) are presented in the lower panel. Eels with the closest scores to the mean of each group (mock; UK ORF35 Del, 100,000 pfu/mL; psoralen/UV-inactivated UK ORF35 Del, 100,000 pfu/mL; and mock-infected with UK Luc) were selected for image illustration.</p>
Full article ">Figure 4 Cont.
<p>Dose-protection effect conferred by UK ORF35 Del in vivo. (<b>A</b>) Flowchart of the experiment. At the time of primary inoculation, yellow eels (28.52 ± 7.30 g, mean ± SD) were mock-infected or infected for 2 h by immersion in water containing the indicated doses of UK ORF35 Del or 100,000 pfu/mL of UK ORF35 Del strain treated by psoralen/UV to inactivate virus infectivity. At 36 days post-primary inoculation, eels were infected for 2 h by immersion in water containing 4000 pfu/mL of UK Luc expressing luciferase as a reporter. At the indicated times post-secondary inoculation, eels (<span class="html-italic">n</span> = 6, consisting of two eels from triplicate tanks) were imaged using IVIS. (<b>B</b>) Average radiance (individual values, mean ± SEM) measured on the entire body surface of fish, i.e., skin (individual values represent the mean values obtained for the left and right sides of each fish), gills (individual values represent the mean values obtained for the left and right gills), brain, heart, and gut–liver, were analyzed by IVIS (<span class="html-italic">n</span> = 6 per timepoint). The average radiance (<span class="html-italic">p</span>(<span class="html-italic">rad</span>)) of each group was compared with the “primary mock-infected group” using a non-parametric Kruskal–Wallis test followed by multiple comparisons with the two-stage step-up method of Benjamini, Krieger, and Yekutieli. Throughout this panel, the data obtained for every individual eel (within each group) are represented by the same symbol to allow for a correlation of the data obtained for the different organs at a specific dpi. The dashed line represents the threshold of positivity, which was calculated by the mean + 3 SD of the values obtained for the mock fish. The number of positive fish among the six analyzed fish is represented by bars (right axis). The positive fish (<span class="html-italic">p</span>(<span class="html-italic">no</span>)) from each group was compared with the primary mock-infected group using the Fisher–Pitman permutation test. <span class="html-italic">p</span> values are represented as follows: ns, not significant; *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01. (<b>C</b>) Representative images of IVIS data (skin, gills, and brain) are presented in the lower panel. Eels with the closest scores to the mean of each group (mock; UK ORF35 Del, 100,000 pfu/mL; psoralen/UV-inactivated UK ORF35 Del, 100,000 pfu/mL; and mock-infected with UK Luc) were selected for image illustration.</p>
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5 pages, 7671 KiB  
Interesting Images
Intrauterine Adhesion-Induced Septated Amniotic Cavity: Ultrasonographic Findings in Second and Third Trimesters
by Jo-Ting Huang, Yu-Ming Chen, Ching-Chang Tsai, Hsin-Hsin Cheng, Yun-Ju Lai, Pei-Fang Lee, Te-Yao Hsu and Kun-Long Huang
Diagnostics 2024, 14(24), 2826; https://doi.org/10.3390/diagnostics14242826 - 16 Dec 2024
Viewed by 395
Abstract
A 40-year-old woman who had obstetric history of one vaginal delivery and two surgical abortions to terminate early pregnancy received regular prenatal care without any systemic maternal diseases. During the detailed second trimester ultrasound, a homogenous adhesion-induced pseudocystic lesion of 8.6 × 7.4 [...] Read more.
A 40-year-old woman who had obstetric history of one vaginal delivery and two surgical abortions to terminate early pregnancy received regular prenatal care without any systemic maternal diseases. During the detailed second trimester ultrasound, a homogenous adhesion-induced pseudocystic lesion of 8.6 × 7.4 cm was found between the inlet of the endocervix and the uterine cavity in the lower segment of the uterus. There was a clear septum with an inlet of about 2.6 cm near the right lower segment of the uterus. Transvaginal sonography showed a cervical length of 3.29 cm without dilatation. No gross fetal anomalies were found. Sometimes, the fetal head or limbs moved into this cystic space. At 36 3/7 weeks of gestation, a cesarean section was arranged for fetal breech presentation and pre-labor rupture of the membrane. After the delivery of the baby and its placenta, there was no obvious septum in the uterine cavity but only a very short fibrous tissue from the posterior wall of uterus, which could be destroyed when the baby was delivered. No adverse outcomes for the mother or the neonate were observed. Full article
(This article belongs to the Special Issue Imaging for the Diagnosis of Obstetric and Gynecological Diseases)
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<p>Transabdominal ultrasound revealed an adhesion-induced pseudocyst measuring 8.6 × 7.4 cm located between the endocervical inlet and uterine cavity in the lower uterine segment (As shown by the dotted line distance). The inlet was identified at the 10 o’clock position of the pseudocyst. The longest inlet of the adhesion-induced pseudocyst measured 2.6 cm (<a href="#diagnostics-14-02826-f002" class="html-fig">Figure 2</a>). Uterine septa were present on anterior and posterior walls, resulting from intrauterine adhesions (<a href="#diagnostics-14-02826-f003" class="html-fig">Figure 3</a> and <a href="#diagnostics-14-02826-f004" class="html-fig">Figure 4</a>). Transvaginal ultrasound showed cervical length of 3.29 cm without dilation, excluding cervical incompetence (CI) (<a href="#diagnostics-14-02826-f005" class="html-fig">Figure 5</a>). Detailed second-trimester ultrasound detected no gross fetal anomalies or bulging uterine diverticulum. Fetal limbs and head moved freely into and out of the adhesion-induced pseudocavity without hyperechoic band-like lesions, making amniotic band syndrome unlikely. At 36 3/7 weeks’ gestation, cesarean delivery was performed due to pre-labor rupture of membranes and fetal breech presentation. Post-placental delivery examination revealed only a short fibrous tissue extending from the posterior uterine wall (<a href="#diagnostics-14-02826-f006" class="html-fig">Figure 6</a>). The adhesion-induced septa may have been disrupted during fetal delivery. No adverse maternal or neonatal outcomes were observed.</p>
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<p>The maximal diameter of the pseudocyst’s opening measured 2.69 cm (As shown by the dotted line distance).</p>
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<p>Uterine septa were present on anterior and posterior walls, resulting from intrauterine adhesions (As shown by the white arrows).</p>
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<p>When the ultrasound probe was moved to the right side of the uterus, the adhesion-induced septa were visualized perpendicular to the anterior and posterior uterine walls, without inlet (As shown by the white arrows).</p>
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<p>Transvaginal sonographic assessment demonstrated a cervical length measurement of 3.29 cm (As shown by the dotted line distance), with no evidence of cervical dilation.</p>
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<p>Examination after placental delivery revealed only a small strand of fibrous tissue extending from the posterior uterine wall (As shown by the yellow arrow). Dilation and curettage (D&amp;C) is a common surgical procedure for managing pathogenic gestational tissue or elective pregnancy termination. In non-pregnant patients, it also serves as a therapeutic and diagnostic tool for abnormal uterine bleeding [<a href="#B1-diagnostics-14-02826" class="html-bibr">1</a>,<a href="#B2-diagnostics-14-02826" class="html-bibr">2</a>,<a href="#B3-diagnostics-14-02826" class="html-bibr">3</a>]. Although D&amp;C is generally considered a safe procedure, it carries potential complications including cervical or uterine bleeding, infection, uterine perforation, and intrauterine adhesions [<a href="#B4-diagnostics-14-02826" class="html-bibr">4</a>,<a href="#B5-diagnostics-14-02826" class="html-bibr">5</a>,<a href="#B6-diagnostics-14-02826" class="html-bibr">6</a>]. Notably, injury to the decidua basalis leading to endometrial fibrous scarring and fusion of opposing surfaces is believed to be a possible mechanism for intrauterine adhesion formation [<a href="#B7-diagnostics-14-02826" class="html-bibr">7</a>]. Studies have also shown that recurrent pregnancy loss is associated with intrauterine adhesions, with evidence suggesting that hysteroscopic surgery may improve pregnancy outcomes [<a href="#B8-diagnostics-14-02826" class="html-bibr">8</a>].This series of images warrants discussion regarding the distinction among intrauterine adhesions, uterine diverticulum (UD), and CI during pregnancy. UD is an uncommon condition that may be either secondary to intervention/trauma or primarily developed [<a href="#B9-diagnostics-14-02826" class="html-bibr">9</a>]. It is usually discovered incidentally on sonography, appearing as a cystic lesion adjacent to or arising from the uterus, with walls comprising myometrium and a cavity communicating with the uterine lumen [<a href="#B10-diagnostics-14-02826" class="html-bibr">10</a>]. Primary UD is an extremely rare anomaly resulting from the failed midline fusion of the Müllerian duct during final uterine development. Weak points in the uterine wall may dilate during pregnancy and labor, forming a diverticulum [<a href="#B11-diagnostics-14-02826" class="html-bibr">11</a>]. Primary UD typically presents with symptoms including abnormal uterine bleeding and dysmenorrhea. Secondary UD is an iatrogenic condition that develops after uterine intervention or trauma. It may lead to abnormal placental attachment disorders, such as placenta accreta spectrum or ectopic pregnancy [<a href="#B12-diagnostics-14-02826" class="html-bibr">12</a>]. Ultrasound screening for the thinning of the uterine segment is crucial; however, this sign was not present in our case. UD can be misdiagnosed as degenerating uterine myoma, adenomyotic cyst, uterine malformations (such as unicornuate uterus, bicornuate uterus with single cervix, and incomplete septate uterus), or adnexal cyst based on sonographic appearance [<a href="#B13-diagnostics-14-02826" class="html-bibr">13</a>,<a href="#B14-diagnostics-14-02826" class="html-bibr">14</a>,<a href="#B15-diagnostics-14-02826" class="html-bibr">15</a>,<a href="#B16-diagnostics-14-02826" class="html-bibr">16</a>]. Another differential diagnosis is CI, defined as cervical length &lt; 25 mm before 24 weeks’ gestation on transvaginal ultrasound, which provides an accurate measurement of the maximum closed cervical canal length [<a href="#B17-diagnostics-14-02826" class="html-bibr">17</a>]. Cervical funneling, characterized by a dilated endocervical canal with protruding fetal membranes, fetal parts, or umbilical cord, is a more reliable indicator [<a href="#B18-diagnostics-14-02826" class="html-bibr">18</a>]. Fundal pressure during transvaginal sonography may aid early CI detection in symptomatic women [<a href="#B19-diagnostics-14-02826" class="html-bibr">19</a>].</p>
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117 pages, 11215 KiB  
Conference Report
Abstracts of the 16th European Congress of Paediatric and Adolescent Gynaecology
by Panagiotis Christopoulos, Anastasia Vatopoulou, Lina Michala, Zuzana Nižňanská, Zoran Stankovic, Evelien Roos, Theodoros Theodoridis, Pandelis Tsimaris, Pallavi Lathe, Angelos Daniilidis, Nikos F. Vlahos and Žana Bumbulienė
J. Clin. Med. 2024, 13(24), 7574; https://doi.org/10.3390/jcm13247574 - 12 Dec 2024
Viewed by 283
Abstract
Objectives of Paediatric and Adolescent Gynaecology (PAG): PAG aims to foster a collaborative environment that bridges knowledge from various disciplines to ensure the highest quality of care for children and adolescents with gynaecological issues. The European Association of PAG and HellenicPAG, like all [...] Read more.
Objectives of Paediatric and Adolescent Gynaecology (PAG): PAG aims to foster a collaborative environment that bridges knowledge from various disciplines to ensure the highest quality of care for children and adolescents with gynaecological issues. The European Association of PAG and HellenicPAG, like all National PAG Societies, support research and education to advance new insights, improve health outcomes, enhance quality of life, and protect future fertility. Additionally, PAG promotes international cooperation by proposing guidelines for good clinical practice in terms of prevention, diagnosis, procedures, and treatment. Scientific Program and Highlights: The scientific committee curated a comprehensive program featuring renowned experts and researchers from across Europe. Over four days, participants explored a wide range of topics through lectures, panel discussions, case studies, debates, video presentations, and workshops. The conference attracted professionals from various disciplines, including gynaecology, paediatrics, paediatric endocrinology, psychology, plastic surgery, and paediatric surgery. The collaboration between the European Association of Paediatric and Adolescent Gynaecology and the Hellenic Society of Paediatric and Adolescent Gynaecology resulted in a cutting-edge scientific program. World-renowned experts (Appendix A) presented on all aspects of paediatric and adolescent gynaecology, with a particular focus on emerging topics such as abortion, global rights, transgender care, teenage pregnancy, and more. Theme—Individualized Care in an Evolving World: The congress theme aligned perfectly with the evolving landscape of PAG. By presenting high-quality, evidence-based topics, the program aimed to equip the next generation of specialists with the knowledge and skills needed to provide individualized care in a rapidly changing world. Full article
(This article belongs to the Collection Pediatric and Adolescent Gynecology)
Show Figures

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<p>Abdominal ultrasound.</p>
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<p>A nonlatex 22 French foley catheter placed in the vagina.</p>
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<p>Placement of the stent after 3 months.</p>
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<p>Summary of the study design.</p>
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<p>(<b>a</b>). External genitalia. (<b>b</b>) A normal clitoris after pulling the excess prepuce skin.</p>
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<p>The number of girls in each age group is shown.</p>
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<p>(<b>a</b>) Word clouds to the question “What is TS for you?” during the first meeting, (<b>b</b>) Word clouds to the question “What are the obstacles you are facing in TS” during the first meeting, (<b>c</b>) Word clouds to the question “What is TS for you?” during the last meeting, (<b>d</b>) Word clouds to the question “What did you take with you from the support group?” during the last meeting.</p>
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<p>Large polyp-like mass protruding from the vaginal introitus and otherwise normal external genitalia.</p>
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<p>Laparoscopic right salpingo-oophorectomy.</p>
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<p>Ultrasound and MRI of cases with OHVIRA.</p>
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<p>Septum resection for OHVIRA.</p>
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<p>Ultrasound examination.</p>
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<p>MRI.</p>
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<p>Breast Asymmetry.</p>
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<p>Excised fibroadenoma specimen.</p>
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<p><b>14-year-old teenage girl with primary amenorrhea, pelvic pain.</b> (<b>A</b>) Sagittal Τ2W MR image of the pelvis demonstrates absence of a normal uterus and cervix (arrow). In the expected location of the vaginal canal (v) a thin fatty band is observed. (<b>Β</b>) Axial oblique Τ2W MR image of the pelvis in the same patient shows a left uterine remnant (arrow). A distended with hemorrhagic fluid endometrial cavity (asterisk) and associated hematosalpinx (arrowheads) are observed, indicative of the presence of functional endometrium. Closely associated normal volume ipsilateral ovary (o). (<b>C</b>) Axial Τ2W MR image of the upper abdomen in the same patient shows absence of the right kidney. Normal renal location and hypertrophy is shown on the left (arrow). Findings are consistent with Müllerian agenesis with unilateral uterine remnant with functional endometrium.</p>
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<p><b>18-year-old teenage girl with pelvic pain.</b> (<b>A</b>) Axial oblique Τ2W MR image of the pelvis shows a left banana-shaped hemiuterus and single cervix (arrows). There is a closely associated right blind rudimentary horn (asterisk). (<b>Β</b>) Axial oblique Τ2W MR image of the pelvis at an upper level demonstrates the right rudimentary horn distended with menstrual fluid (asterisk). (<b>C</b>) Coronal oblique Τ2W MR image of the pelvis in the same patient shows associated right hematosalpinx (arrows) and free pelvic fluid (asterisk). Findings are consistent with left unicornuate uterus with right associated non-communicating uterine remnant with functional endometrium.</p>
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<p>A 20-years-old girl with painful and palpable mass in her vulva.</p>
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<p>Created with BioRender.com.</p>
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<p>QR code.</p>
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<p>Moos’s Menstrual distress Questionnair.</p>
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<p>Intraoperative image of the patient with serous cystadenoma.</p>
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<p>Intraoperative image of the patient with serous cystadenoma.</p>
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