Dermatology residents' perspectives on virtual dermatopathology education.

BACKGROUND: Dermatopathology education accounts for 30% of U.S. dermatology residency training. The COVID-19 pandemic expedited the implementation of virtual dermatopathology in place of traditional microscopy for resident education. This study examined U.S. dermatology residents' perceptions of virtual dermatopathology, as research in this area is lacking. METHODS: An anonymous, confidential, institutional review board-approved survey was electronically distributed to U.S. dermatology residents consisting of 16 questions comparing attitudes towards virtual and traditional dermatopathology education. Responses were n = 59. Statistical analysis was performed using SAS software. RESULTS: Participants believe virtual imaging is superior to conventional microscopy in schedule flexibility (96.6% vs. 1.7%, p < 0.0001), lecture convenience (94.8% vs. 0.0%, p < 0.0001), personal review (96.6% vs. 0.0%, p < 0.0001), cost-effectiveness (64.4% vs. 6.8%, p < 0.0001), and board exam preparation (52.5% vs. 16.9%, p = 0.0005). Conventional microscopy was favored for image quality (50.8% vs. 25.4%, p = 0.0127) and overall utility (50.8% vs. 27.1%, p = 0.0195). CONCLUSIONS: Our study supports virtual dermatopathology utilization as a valuable tool in dermatology residency training. Also it is shown that conventional microscopy training continues to play a key role. Further studies should examine whether, if ever, virtual dermatopathology could gradually replace conventional microscopy with the advent of newer and more powerful digital and scanning technology.

Co-circulation and genetic characteristics of porcine circoviruses in postweaning multisystemic wasting syndrome cases in commercial swine farms.

This study aimed to investigate the co-infection and genetic characteristics of Porcine circoviruses in PMWS-affected pigs in five commercial farrow-to-finish swine farms in Vietnam. By the end of 2022, the percentage of PMWS-affected pigs in these farms has increased significantly compared to previous years. The lymph node samples from ten PMWS typical cases were randomly collected to test for the presence of PRRSV, PCV2, PCV3 and PCV4. While PRRSV and PCV4 were not found in these cases, 10 and 3 out of 10 samples were positive for PCV2 and PCV3, respectively. Three farms in the study showed the co-infection of PCV2 and PCV3 in affected pigs. Besides, all PCV-positive samples were sequenced to evaluate genetic characterization of PCVs in PMWS-affected cases. Phylogenetic analysis showed that all PCV3 strains in the study were clustered into PCV3b genotype. 8 out of 10 PCV2 strains belonged to PCV2d genotype while the remaining two strains belonged to PCV2b genotypes. Two farms had co-circulation of PCV2b and PCV2d genotypes in two different age groups of pigs, which is reported for the first time in Vietnam. Several amino acid substitutions were identified in important antigenic regions in the capsid protein of the PCV2 field strains compared to vaccine strains. Taken together, the results showed the high co-prevalence of PCV3 and PCV2, and the wide genetic diversity of PCV2 field and vaccine strains may be the cause of the increased PMWS situation in these pig farms. Supplementary Information: The online version contains supplementary material available at 10.1007/s13337-023-00849-4.

Development of optimized protocol for culturing African swine fever virus field isolates in MA104 cells.

The goal of this study was to identify a candidate commercial cell line for the replication of African swine fever virus (ASFV) by comparing several available cell lines with various medium factors. In the sensitivity test of cells, MA104 and MARC-145 had strong potential for ASFV replication. Next, MA104 cells were used to compare the adaptation of ASFV obtained from tissue homogenates and blood samples in various infectious media. At the 10th passage, the ASFV obtained from the blood sample had a significantly higher viral load than that obtained from the tissue sample (P = 0.000), exhibiting a mean cycle threshold (Ct) value = 20.39 ± 1.99 compared with 25.36 ± 2.11. For blood samples, ASFV grew on infectious medium B more robustly than on infectious medium A (P = 0.006), corresponding to a Ct value = 19.58 ± 2.10 versus 21.20 ± 1.47. African swine fever virus originating from blood specimens continued to multiply gradually and peaked in the 15th passage, exhibiting a Ct value = 14.36 ± 0.22 in infectious medium B and a Ct value = 15.42 ± 0.14 in infectious medium A. When ASFV was cultured from tissue homogenates, however, there was no difference (P = 0.062) in ASFV growth between infectious media A and B. A model was developed to enhance ASFV replication through adaptation to MA104 cells. The lack of mutation at the genetic segments encoding p72, p54, p30, and the central hypervariable region (CVR) in serial culture passages is important in increasing the probability of maintaining immunogenicity when developing a vaccine candidate.

L'objectif de cette étude était d'identifier une lignée cellulaire commerciale candidate pour la réplication du virus de la peste porcine africaine (ASFV) en comparant plusieurs lignées cellulaires disponibles et différents milieux. Lors du test de sensibilité des cellules, MA104 et MARC-145 présentaient un fort potentiel pour la réplication d'AFSV. Par la suite, les cellules MA104 ont été utilisées pour comparer l'adaptation d'ASFV obtenu d'homogénats de tissus et d'échantillons de sang dans différents milieux. Au dixième passage, l'ASFV obtenu de l'échantillon de sang avait une charge virale significativement plus élevée que celle obtenue de l'échantillon de tissu (P = 0,000), avec une valeur seuil moyenne de cycles (Ct) de 20,39 ± 1,99 comparativement à 25,36 ± 2,11. Pour les échantillons sanguins, l'ASFV a poussé sur le milieu B de manière plus robuste que sur le milieu A (P = 0,006), ce qui correspond à une valeur Ct de 19,58 ± 2,10 versus 21,20 ± 1,47. L'ASFV provenant des échantillons sanguins continua de se multiplier graduellement et atteignit un pic au 15e passage, avec une valeur Ct de 14,36 ± 0,22 dans le milieu B et une valeur Ct de 15,42 ± 0,14 dans le milieu A. Toutefois, lorsque l'ASFV fut cultivé à partir des homogénats de tissus, il n'y avait pas de différence (P = 0,062) dans la croissance d'ASFV entre les milieux A et B. Un modèle a été développé pour augmenter la réplication d'ASFV par adaptation aux cellules MA104. L'absence de mutation au segment génétique codant pour p72, p54, p30, et la région hypervariable centrale (CVR) dans des passages en série en culture est importante en augmentant la probabilité de maintenir une immunogénicité lors du développement d'un vaccin candidat.(Traduit par Docteur Serge Messier).

Genetic Characterization of African Swine Fever Virus in Various Outbreaks in Central and Southern Vietnam During 2019-2021.

This study aimed to identify potential genetic diversity among African swine fever virus (ASFV) strains circulating in central and southern Vietnam. Thirty ASFV strains were collected from domestic pigs and convalescent pigs with ASFV-infected clinical signs from 19 different provinces of central and southern Vietnam during 2019-2021. A portion of the B646L (p72) gene and the entire E183L (p54), CP204L (p30), and B602L (CVR) genes were amplified, purified, and sequenced. Web-based BLAST and MEGA X software were used for sequence analysis. Analysis of the partial B646L (p72) gene, the full-length E183L (p54) and CP204L (p30) genes, and the central hypervariable region (CVR) of the B602L gene sequence showed that all 30 ASFV isolates belonged to genotype II and were 100% identical to the previously identified strains in Vietnam and China. Analysis of the p72, p54, and p30 regions did not indicate any change in the nucleotide and amino acid sequences among these strains in 3 years of research. No novel variant was found in the CVR within the B602L gene. Analysis of the CVR showed that these ASFV strains belong to subgroup XXXII. The results of this study revealed that these ASFVs shared high similarity with ASFV isolates detected previously in northern Vietnam and China. Taken together, the results of this study and a previous study in Vietnam showed high stability and no genetic diversity in the ASFV genome.

Long-term follow-up of convalescent pigs and their offspring after an outbreak of acute African swine fever in Vietnam.

African swine fever (ASF) is a contagious haemorrhagic disease in pigs and has become endemic in several Vietnam provinces since the first outbreak in 2019. The presence of carriers and the recurrence of disease in the surviving swine herd after an ASF outbreak has not previously been properly evaluated. In this study, pigs naturally infected with an acute form of ASF were allowed to recover from the disease. A serological follow-up was conducted for more than 14 months with 14 convalescent gilts and their offspring. All convalescent animals had long lasting high serum antibody levels without persistent viremia. They also did not excrete virus via nasal discharge post-recovery. These convalescent pigs could partially perform as replacement gilts despite the fact that ASF affected reproductive performance. Here, we confirmed that there were neither the carriers of nor recurrence of disease in the convalescent pigs and their offspring following the outbreak of acute ASF. These findings may facilitate efforts to design a new farming model in ASF endemic provinces in Vietnam where there is a lack of a repopulation strategy due to the limited funding received from the local regulatory authorities.

Is genital tract infection related to tubal diseases in infertile Vietnamese women?

INTRODUCTION: The goal of this study was to identify the profile of genital tract infections and their relationship with clinical and demographic parameters as well as tubal diseases among infertile women in Vietnam. METHODOLOGY: In this cross-sectional descriptive study, we enrolled 597 women undergoing infertility treatment at the Center for Reproductive Endocrinology and Infertility, Hue University Hospital, Vietnam. All of the study participants were interviewed and examined by a gynecologist. Consecutive tests were then conducted including direct microscopy examination (wet mount and Gram stain), vaginal culture, polymerase chain reaction (PCR) for chlamydia diagnosis from a cervical canal swab, and a blood test for syphilis detection. A hysterosalpingogram (HSG) was carried out to examine the uterine cavity and Fallopian tubes. RESULTS: A gynecologic infection was diagnosed in 43.4% (259/597) of the infertile women. Bacterial vaginosis was the most common condition at 19.6%of the cases. Candida spp., Chlamydia trachomatis, and Trichomonas vaginalis infections accounted for 17.4%, 3.7%, and 0.3%, respectively. Normal HSG results accounted for 87.4% of the women while 5.5% had 2-sided tubal occlusions, 5.4% had 1-sided tubal occlusions, 1.0% had 1-sided hydrosalpinx, and 0.7% had 2-sided hydrosalpinx. There was no significant association between tubal diseases and current infections; however, aerobic vaginitis increased the risk of tubal diseases by 2.4 times. CONCLUSIONS: A marked proportion of infertile Vietnamese women have genital tract infections that can significantly influence their reproductive function and performance. These infections should be routinely screened and treated properly to prevent their consequences, such as infertility, which is especially important in developing countries.

Emergence of unusual vanA/vanB2 genotype in a highly mutated vanB2-vancomycin-resistant hospital-associated E. faecium background in Vietnam.

Enterococcus faecium has become a globally disseminated nosocomial pathogen mainly because of acquisition and diffusion of virulence factors and multidrug resistance determinants, including glycopeptides, which are some of the last resort antimicrobials used to treat more serious infections common in high-risk patients. In this study we investigated and characterized hospital-associated (HA) E. faecium isolates collected at Hue Central Hospital, Vietnam. Our results highlighted the spread among hospital wards of a surprisingly heterogeneous multidrug-resistant E. faecium population comprising five different CC17-related sequence types (STs), of which 46% VREf carry the vanB gene. Whole genome sequencing of selected E. faecium isolates showed that VREf from different STs carried the same chromosomal integrated Tn1549-like transposon, with a highly mutated vanB2-operon, showing an increased level of vancomycin resistance (VanB phenotype) and able, in one isolate, to confer resistance to teicoplanin (VanA incongruent phenotype). Two unusual vanA/vanB2-type strains were detected within the vanB2-type ST17 population, harbouring a Tn1546-vanA-like transposon in pJEG40-like plasmids. Wg-SNPs-based analysis showed the genetic relatedness of VSEf/VREf of the same STs and indicated lateral exchange of the Tn1549-like element among isolates followed by clonal expansion. Microevolution among ST17 isolates, including the vanA/vanB2-type strains, and inter-wards VREf transmission, were highlighted. The use of teicoplanin is strongly discouraged in the study hospital because of the spreading of Tn1549-vanB2 associated to teicoplanin resistance. A rational use of glycopeptides and effective surveillance measures are required to reduce nosocomial VSEF/VREf spread and to avoid the rise of unusual and misleading VREf genotypes.

Cell-Based High-Throughput Luciferase Reporter Gene Assays for Identifying and Profiling Chemical Modulators of Endoplasmic Reticulum Signaling Protein, IRE1.

Endoplasmic reticulum (ER) stress activates three distinct signal transducers on the ER membrane. Inositol-requiring protein 1 (IRE1), the most conserved signal transducer, plays a key role in ER stress-mediated signaling. During ER stress, IRE1 initiates two discrete signaling cascades: the "adaptive" signaling cascade mediated by the XBP1 pathway and the "alarm" signaling cascade mediated by stress-activated protein kinase pathways. Fine-tuning of the balance between the adaptive and alarm signals contributes significantly to cellular fate under ER stress. Thus, we propose that the design of high-throughput screening (HTS) assays to selectively monitor IRE1 mediated-signaling would be desirable for drug discovery. To this end, we report the generation of stable human neural cell lines and development of cell-based HTS luciferase (Luc) reporter gene assays for the identification of pathway-specific chemical modulators of IRE1. We implemented a cell-based Luc assay using a chimeric CHOP-Gal4 transcription factor in 384-well format for monitoring IRE1 kinase-mediated p38MAPK activation and an unfolded response pathway element (URPE)-Luc cell-based assay in 1536-well format for monitoring IRE1's RNase-mediated activation of XBP1. Chemical library screening was successfully conducted with both the CHOP/Gal4-Luc cells and UPRE-Luc engineered cells. The studies demonstrate the feasibility of using these HTS assays for discovery of pathway-selective modulators of IRE1.

TR-FRET-based high-throughput screening assay for identification of UBC13 inhibitors.

UBC13 is a noncanonical ubiquitin conjugating enzyme (E2) that has been implicated in a variety of cellular signaling processes due to its ability to catalyze formation of lysine 63-linked polyubiquitin chains on various substrates. In particular, UBC13 is required for signaling by a variety of receptors important in immune regulation, making it a candidate target for inflammatory diseases. UBC13 is also critical for double-strand DNA repair and thus a potential radiosensitizer and chemosensitizer target for oncology. The authors developed a high-throughput screening (HTS) assay for UBC13 based on the method of time-resolved fluorescence resonance energy transfer (TR-FRET). The TR-FRET assay combines fluorochrome (Fl)-conjugated ubiquitin (fluorescence acceptor) with terbium (Tb)-conjugated ubiquitin (fluorescence donor), such that the assembly of mixed chains of Fl- and Tb-ubiquitin creates a robust TR-FRET signal. The authors defined conditions for optimized performance of the TR-FRET assay in both 384- and 1536-well formats. Chemical library screens (total 456 865 compounds) were conducted in high-throughput mode using various compound collections, affording superb Z' scores (typically >0.7) and thus validating the performance of the assays. Altogether, the HTS assays described here are suitable for large-scale, automated screening of chemical libraries in search of compounds with inhibitory activity against UBC13.