Molecular epidemiology of SARS-CoV-2 isolated from COVID-19 family clusters.

BACKGROUND: Transmission within families and multiple spike protein mutations have been associated with the rapid transmission of SARS-CoV-2. We aimed to: (1) describe full genome characterization of SARS-CoV-2 and correlate the sequences with epidemiological data within family clusters, and (2) conduct phylogenetic analysis of all samples from Yogyakarta and Central Java, Indonesia and other countries. METHODS: The study involved 17 patients with COVID-19, including two family clusters. We determined the full-genome sequences of SARS-CoV-2 using the Illumina MiSeq next-generation sequencer. Phylogenetic analysis was performed using a dataset of 142 full-genomes of SARS-CoV-2 from different regions. RESULTS: Ninety-four SNPs were detected throughout the open reading frame (ORF) of SARS-CoV-2 samples with 58% (54/94) of the nucleic acid changes resulting in amino acid mutations. About 94% (16/17) of the virus samples showed D614G on spike protein and 56% of these (9/16) showed other various amino acid mutations on this protein, including L5F, V83L, V213A, W258R, Q677H, and N811I. The virus samples from family cluster-1 (n = 3) belong to the same clade GH, in which two were collected from deceased patients, and the other from the survived patient. All samples from this family cluster revealed a combination of spike protein mutations of D614G and V213A. Virus samples from family cluster-2 (n = 3) also belonged to the clade GH and showed other spike protein mutations of L5F alongside the D614G mutation. CONCLUSIONS: Our study is the first comprehensive report associating the full-genome sequences of SARS-CoV-2 with the epidemiological data within family clusters. Phylogenetic analysis revealed that the three viruses from family cluster-1 formed a monophyletic group, whereas viruses from family cluster-2 formed a polyphyletic group indicating there is the possibility of different sources of infection. This study highlights how the same spike protein mutations among members of the same family might show different disease outcomes.

Functional outcomes of patients with short-segment Hirschsprung disease after transanal endorectal pull-through.

BACKGROUND: Transanal endorectal pull-through (TEPT) is considered the most preferable treatment method for Hirschsprung disease (HSCR) since it is less invasive and has fewer morbidities than transabdominal pull-through. Here, functional outcomes in short-segment HSCR patients after TEPT were assessed and associated with the prognostic factors. METHODS: Krickenbeck classification was used to assess the functional outcomes in patients with HSCR after TEPT surgery at our institution from 2012 to 2020. RESULTS: Fifty patients were involved in this study. Voluntary bowel movement (VBM) was achieved in 82% of subjects. Nine (18%) subjects had soiling grade 1, while two (4%) and two (4%) patients suffered constipation that was manageable with diet and laxative agents, respectively. Patients who underwent TEPT at ≥ 4 years old tended to have soiling more than patients who underwent TEPT at < 4 years old (OR = 16.47 [95% CI 0.9-301.61]; p = 0.06), whereas patients with post-operative complications had 10.5-fold higher risk for constipation than patients without post-operative complications (p = 0.037; 95% CI 1.15-95.92). Multivariate analysis showed male sex was significantly associated with VBM (OR = 9.25 [95% CI 1.34-63.77]; p = 0.024), while post-operative complications were strongly correlated with constipation (OR = 10 [95% CI 1.09-91.44]; p = 0.04). CONCLUSIONS: The functional outcomes of HSCR patients after TEPT in our institution are considered relatively good. Moreover, the VBM, soiling, and constipation risk after TEPT might be affected by sex, age at TEPT performed, and post-operative complications, respectively, while the age at TEPT performed might not be associated with functional outcomes. Further multicenter studies with a larger sample size are necessary to clarify and confirm our findings.

Histopathological findings for prediction of liver cirrhosis and survival in biliary atresia patients after Kasai procedure.

BACKGROUND: Without early recognition and Kasai procedure, biliary atresia (BA) results in liver cirrhosis and leads to either transplantation or death at a young age. We aimed to characterize the liver histopathological findings for prediction of cirrhosis and survival in BA patients after Kasai surgery. METHODS: We retrospectively reviewed all histopathological results for BA patients who underwent liver biopsy during Kasai surgery from August 2012 to December 2018 in Dr. Sardjito Hospital, Yogyakarta, Indonesia. RESULTS: Fifty infants with BA were ascertained in our study, of whom 27 were males and 23 were females. The median age of Kasai procedure was 102.5 days (interquartile range (IQR), 75.75-142.25 days). There were 33 (66%) and 17 (34%) BA patients with and without liver cirrhosis, respectively, while the overall survival was 52%. The patients with a severe bile duct proliferation, severe cholestasis, and severe portal inflammation have a higher risk by 27-, 22-, and 19.3-fold, respectively, to develop liver cirrhosis compared with patients with a moderate/mild bile duct proliferation, moderate/mild/without cholestasis, and moderate/mild portal inflammation, respectively (p = 3.6 × 10- 6, 5.6 × 10- 4, and 1.6 × 10- 3, respectively), while the giant cell transformation was not associate with the development of liver cirrhosis (p = 0.77). The bile duct proliferation was strongly correlated with cholestasis and portal inflammation (p = 7.3 × 10- 5 and 2 × 10- 4, respectively), and cholestasis was also significantly correlated with portal inflammation (p = 0.016). Interestingly, the age at Kasai procedure was strongly associated with the development of liver cirrhosis (p = 0.02), but not with the patients' survival (p = 0.33), while the degree of fibrosis and cholestasis were significantly correlated with the patients' survival, with HR of 3.9 (95% CI = 1.7-9.0; p = 0.017) and 3.1 (95% CI = 1.4-7.0; p = 0.016), respectively. CONCLUSIONS: Histopathological findings of bile duct proliferation, cholestasis, and portal inflammation can predict the liver cirrhosis development in patients with BA. Furthermore, degree of fibrosis and cholestasis affect the patients' survival following the Kasai operation.

Anorectal malformation patients' outcomes after definitive surgery using Krickenbeck classification: A cross-sectional study.

BACKGROUND: The survival of anorectal malformation (ARM) patients has been improved in the last 10 years because of the improvement in management of neonatal care and surgical approaches for ARM patients. Thus, the current management of ARM patients are focusing on the functional outcomes after definitive surgery. Here, we defined the type of ARM and assessed the functional outcomes, including voluntary bowel movement (VBM), soiling, and constipation, in our patients following definitive surgery using Krickenbeck classification. METHODS: We conducted a cross-sectional study to retrospectively review medical records of ARM patients who underwent a definitive surgery at Dr. Sardjito Hospital, Indonesia, from 2011 to 2016. RESULTS: Forty-three ARM patients were ascertained in this study, of whom 30 males and 13 females. Most patients (83.7%) were normal birth weight. There were ARM without fistula (41.9%), followed by rectourethral fistula (25.5%), perineal fistula (18.6%), vestibular fistula (9.3%), and rectovesical fistula (4.7%). The VBM was achived in 53.5% patients, while the soiling and constipation rates were 11.6% and 9.3%, respectively. Interestingly, patients with normal birth weight showed higher frequency of VBM than those with low birth weight (OR = 9.4; 95% CI = 1.0-86.9; p = 0.04), while male patients also had better VBM than females (OR = 3.9; 95% CI = 1.0-15.6) which almost reached a significant level (p = 0.09). However, VBM was not affected by ARM type (p = 0.26). Furthermore, there were no significant associations between gender, birth weight, and ARM type with soiling and constipation, with p-values of 1.0, 1.0, and 0.87; and 0.57, 1.0, and 0.94, respectively. CONCLUSIONS: Functional outcomes of ARM patients in our hospital are considered relatively good with more than half of children showing VBM and only relatively few patients suffering from soiling and constipation. The frequency of VBM might be associated with birth weight and gender, but not ARM type, while the soiling and constipation did not appear to be correlated with birth weight, gender, nor ARM type. Further multicenter study is necessary to compare our findings with other centers.

Aberrant UBR4 expressions in Hirschsprung disease patients.

BACKGROUND: Recently, pathogenic alleles within ubiquitin N-recognin domain-containing E3 ligase 4 (UBR4) gene have been shown to be associated with Hirschsprung disease (HSCR). We determined the UBR4 expressions in Indonesian HSCR patients. METHODS: We analyzed the UBR4 expressions in the colons of HSCR patient and anorectal malformation (ARM) patient as control by real-time polymerase chain reaction (qPCR). RESULTS: Thirty-seven patients with non-syndromic HSCR and eighteen controls were involved in this study. qPCR revealed that the UBR4 expression was strongly decreased (0.77-fold) in the ganglionic group of patients with HSCR compared to the control group with ARM (ΔCT 2.43 ± 0.36 vs. 2.05 ± 0.69; p = 0.009), whereas the UBR4 expression was also significantly reduced (0.79-fold) in the aganglionic group of patients with HSCR compared to the control group with ARM (ΔCT 2.39 ± 0.46 vs. 2.05 ± 0.69; p = 0.044). However, the UBR4 expression change was not associated with gender (p = 0.35 and 0.80), nor with degree of aganglionosis both in ganglionic and aganglionic colons (p = 0.72 and 0.73), respectively. CONCLUSION: Our study demonstrates that expression of UBR4 is decreased in both aganglionic and ganglionic colon of HSCR patients.

Neglected orbitozygomaticomaxillary fractures with complications: A case report.

INTRODUCTION: Zygomatic complex fracture is one of the most frequently occuring injuries to facial structures and often involves damage to the surrounding structures, including the orbital floor and/or medial orbital wall. These complicated injuries can cause both serious opthalmic and aesthetic complcations that make reconstructions more difficult. CASE PRESENTATION: A 22 year old male presented with facial asymmetry. On examination, there were neglected fractures of left orbitozygomaticomaxillary complex with various compications. We performed lateral orbitotomy, bone graft, medial and lateral cantopexy with 3D reconstruction models as a guidance. There were some appearance improvements including improved orbital dystopia and corrected facial asymmetry. However, his vision could not be corrected because the reconstruction was too late. DISCUSSION: Several examinations before reconstruction of the injury has to be done, especially radiological examinations to decide proper diagnosis and reconstruction plans. Some complications may make difficulties in establishing precise time for surgery. Three-dimensional (3-D) reconstrucion model can help increase accuracy of the reconstruction but consume more time. CONCLUSION: Severe maxillofacial fracture should be treated immediately to avoid further complications. Multidisciplinary examinations can provide more accurate preliminary recommendations particularly when combine with properly calibrated CT scan imaging.

The effect of 5α-oleandrin on keloid fibroblast activities.

BACKGROUND: Keloids develop due to hyperactivity of keloid fibroblast (KF) in proliferation, migration, and collagen deposition along with low rates of collagen degradation. These are a result of the Wnt/ß catenin signaling pathways under stimulation of TGF-ß. 5α-oleandrin can suppress Wnt-targeted genes of osteosarcoma cells. We aimed to evaluate the anti-fibrotic effects of 5α-oleandrin on KF activities. METHODS: We collected the core of keloid materials from six patients who underwent keloid debulking surgery. Passage 4 of KF cells were then treated with mitomycin-C, 5α-oleandrin, and dilution medium as the negative control. To determine the effective dose of 5α-oleandrin, we diluted 5α-oleandrin into various concentrations. The incubation periods were 24 h, 48 h, and 72 h. The anti-proliferation and anti-fibrotic properties were measured using standard assay. RESULTS: Both the mitomycin-C and 5α-oleandrin treated groups indicated decrease in proliferation index (86.16 ± 4.20% and 73.76 ± 4.94%, respectively), collagen deposition index (90.26 ± 1.72% and 71.35 ± 4.26%, respectively), and migration capacity (33.51 ± 1.50% and 28.57 ± 1.58%, respectively). These were significant changes (p ≤ 0.05) compared to the non-treated group. Antifibrotic activities of 5α-oleandrin in cellular proliferation and collagen deposition were better than mitomycin-C. CONCLUSIONS: The 5α-oleandrin has good antifibrotic effect in keloid fibroblast activities.