ABSTRACT
BACKGROUND: We report changes in the natural history of hip instability with nusinersen treatment among patients with spinal muscular atrophy (SMA) type II after onset of weakness, historically wheelchair-bound but now potentially ambulatory in the era of disease-modifying therapy. METHODS: Patients with genetically confirmed diagnoses of SMA type II who received intrathecal nusinersen from January 1, 2018, to June 30, 2022, were screened for inclusion. Patients with less than 6 months of follow-up, or prior hip surgeries were excluded. Primary clinical outcome measures included scores from Hammersmith motor functional scale expanded (HMFSE), revised upper limb module (RULM), 6-minute walk test (6MWT), and ambulatory status. Radiographic outcomes, including Reimer migration index, the presence of scoliosis, and pelvic obliquity, were also assessed. Secondary outcomes involved comparisons with a historical cohort of SMA type II patients treated at our institution who never received nusinersen. RESULTS: Twenty hips from 5 boys and 5 girls were included in the analysis, with a mean follow-up of 3 years and 8 months. The median age at time of nusinersen initiation was 6.8 years old, ranging between 2.5 and 10.3 years. All patients developed lower limb motor weakness before nusinersen initiation. After treatment with nusinersen, 1 previously stable hip (5%) developed subluxation, 15 hips (75%) remain subluxated, 3 hips (15%) remain dislocated, and 1 hip (5%) remained stable, with a statistically significant difference between the pretreatment and posttreatment groups ( P <0.01). Six patients (60%) were ambulatory at latest follow-up. Six patients (60%) had improved ambulatory ability; 2 had static ambulatory ability (20%); and 2 had deterioration in their walking ability. The median HFMSE score improved from 18.5 (range 0 to 46) to 22 (range 0 to 49) ( P =0.813), whereas the median RULM score improved from 17 (range 2 to 28) to 21.5 (range 5 to 37), which was statistically significant ( P =0.007). CONCLUSIONS: Hip instability persists despite treatment with nusinersen among patients with SMA type II who received nusinersen after onset of lower limb weakness. LEVEL OF EVIDENCE: Therapeutic Level IV.
Subject(s)
Disease Progression , Joint Instability , Muscle Weakness , Oligonucleotides , Spinal Muscular Atrophies of Childhood , Humans , Male , Female , Child , Oligonucleotides/administration & dosage , Oligonucleotides/therapeutic use , Child, Preschool , Muscle Weakness/etiology , Spinal Muscular Atrophies of Childhood/drug therapy , Spinal Muscular Atrophies of Childhood/physiopathology , Joint Instability/drug therapy , Retrospective Studies , Hip Joint/physiopathology , Follow-Up StudiesABSTRACT
PURPOSE: Although it is evident that some patients with adolescent idiopathic scoliosis (AIS) have proprioceptive deficit in peripheral joints, knowledge on the proprioceptive function of the deformed spine is limited. Nonetheless, spinal proprioception in AIS may be affected three-dimensionally, prior studies only focussed on evaluating peripheral proprioception in single plane. Therefore, this study aimed to develop a novel spinal proprioception assessment using three-dimensional motion analysis in patients with AIS. METHODS: Participants were included if they had a primary diagnosis of AIS who did not receive or failed conservative treatments. Three trunk repositioning tests involving flexion-extension, lateral-flexion, and axial-rotation were conducted. A three-dimensional kinematics of the trunk was used as the outcome measures. The proprioceptive acuity was quantified by the repositioning error. The intra-examiner and test-retest reliability were analysed by the intraclass correlation coefficient (ICC). RESULTS: Fifty-nine patients with AIS were recruited. Regarding the trunk flexion-extension test, the single measure ICC showed moderate reliability (0.46) and the average measures ICC demonstrated good reliability (0.72). As for the trunk lateral-flexion test, the reliability of single measure and average measures ICC was moderate (0.44) and good (0.70) reliability, respectively. For the trunk axial-rotation test, the single measure ICC indicated fair reliability (0.32), while the average measures ICC showed moderate reliability (0.59). CONCLUSION: This is the first study to evaluate the reliability of novel three-dimensional spinal proprioception assessments in patients with AIS. The trunk flexion-extension repositioning test may be preferable clinical test given its highest reliability.
Subject(s)
Kyphosis , Scoliosis , Humans , Adolescent , Scoliosis/diagnosis , Reproducibility of Results , Spine , ProprioceptionABSTRACT
BACKGROUND: Cerebral palsy patients are at risk of hip instability, to which various soft tissue and bony surgeries are performed should conservative management fail. We aim to identify factors associated with treatment failure to guide surgical management. METHODS: Cerebral palsy patients treated at 2 university-affiliated tertiary pediatric orthopaedic referral centers with hip stabilization surgery performed for subluxation in 1998 to 2015 with minimum of 5 years follow-up were reviewed. Failure was defined as reoperation to the same hip because of recurrent subluxation. Age, sex, Gross Motor Function Classification System level, tone abnormality, operation type, Reimer's migration index (RMI), and acetabular index (AI) were assessed. Cut-off values were identified through Youden index on receiver operating characteristic curve. RESULTS: Eighty-nine hips from 55 patients with mean follow-up of 12.4 years were analyzed. Revision surgery was performed in 14 hips. Postoperative hip subluxation (P<0.001) and acetabular dysplasia (P=0.001) were predictive of failure, with postoperative RMI conferring an adjusted hazard ratio of 1.13 (95% confidence interval: 1.08-1.19, P<0.001) on multivariable survival analysis. Achieving a postoperative RMI of <27.5% predicts success with 92.9% sensitivity and 72% specificity with area under curve of 0.916 (P<0.001), while postoperative AI of <23.1 degrees predicts success with 92.3% sensitivity and 62.2% specificity with area under curve of 0.796 (P=0.001). In subgroup analysis of soft-tissue-only procedures, RMI >44% preoperative and >32% postoperative were associated with reoperation. In femur-only osteotomies, preoperative RMI >48% and postoperative RMI >28% were associated with failure. In pelvic and combined osteotomies, postoperative RMI >32% and AI >30 degrees were associated with failure. Other factors analyzed were not associated with reoperation. CONCLUSIONS: Patient selection and quality of surgery in terms of residual postoperative hip subluxation and acetabular dysplasia are associated with need for remedial surgery. Soft-tissue-only procedures should aim to correct RMI to <32%. Bony surgery should be considered when preoperative RMI >44%, and pelvic osteotomies if RMI >48%. Pelvic osteotomies should target postoperative RMI <32% and AI <30 degrees. LEVEL OF EVIDENCE: Level II-prognostic study.
Subject(s)
Cerebral Palsy , Hip Dislocation , Acetabulum/diagnostic imaging , Acetabulum/surgery , Cerebral Palsy/complications , Child , Hip Dislocation/surgery , Hip Joint , Humans , Patient Selection , Retrospective Studies , Treatment OutcomeABSTRACT
We determined the susceptibilities of 57 Talaromyces marneffei strains to anidulafungin, itraconazole, voriconazole, and posaconazole with MICs of 2 to 8, 0.002 to 0.004, 0.016 to 0.063, and 0.001 to 0.002 µg/ml by broth microdilution and >32, ≤0.002 to 0.008, ≤0.002 to 0.008, and ≤0.002 µg/ml by Etest, respectively, at yeast phase; MICs at mycelial phase for anidulafungin and posaconazole were 1 to 2 and 0.004 to 0.063 µg/ml, respectively. The results suggest promising activities of posaconazole. Etest can be used for testing of azoles against T. marneffei.
Subject(s)
Antifungal Agents/pharmacology , Echinocandins/pharmacology , Itraconazole/pharmacology , Talaromyces/drug effects , Triazoles/pharmacology , Voriconazole/pharmacology , Anidulafungin , Disk Diffusion Antimicrobial Tests , Humans , Penicillium/drug effects , Penicillium/growth & development , Reagent Strips , Talaromyces/growth & development , Talaromyces/isolation & purificationABSTRACT
Systemic dimorphic fungi cause more than one million new infections each year, ranking them among the significant public health challenges currently encountered. Penicillium marneffei is a systemic dimorphic fungus endemic to Southeast Asia. The temperature-dependent dimorphic phase transition between mycelium and yeast is considered crucial for the pathogenicity and transmission of P. marneffei, but the underlying mechanisms are still poorly understood. Here, we re-sequenced P. marneffei strain PM1 using multiple sequencing platforms and assembled the genome using hybrid genome assembly. We determined gene expression levels using RNA sequencing at the mycelial and yeast phases of P. marneffei, as well as during phase transition. We classified 2,718 genes with variable expression across conditions into 14 distinct groups, each marked by a signature expression pattern implicated at a certain stage in the dimorphic life cycle. Genes with the same expression patterns tend to be clustered together on the genome, suggesting orchestrated regulations of the transcriptional activities of neighboring genes. Using qRT-PCR, we validated expression levels of all genes in one of clusters highly expressed during the yeast-to-mycelium transition. These included madsA, a gene encoding MADS-box transcription factor whose gene family is exclusively expanded in P. marneffei. Over-expression of madsA drove P. marneffei to undergo mycelial growth at 37°C, a condition that restricts the wild-type in the yeast phase. Furthermore, analyses of signature expression patterns suggested diverse roles of secreted proteins at different developmental stages and the potential importance of non-coding RNAs in mycelium-to-yeast transition. We also showed that RNA structural transition in response to temperature changes may be related to the control of thermal dimorphism. Together, our findings have revealed multiple molecular mechanisms that may underlie the dimorphic transition in P. marneffei, providing a powerful foundation for identifying molecular targets for mechanism-based interventions.
Subject(s)
Gene Expression Regulation, Fungal , Penicillium/genetics , Fungal Proteins/genetics , Fungal Proteins/metabolism , Genome, Fungal , MADS Domain Proteins/genetics , Multigene Family , Mycelium/genetics , Penicillium/growth & development , Penicillium/pathogenicity , RNA, Fungal/chemistry , Temperature , Transcription Factors/genetics , TranscriptomeABSTRACT
UNLABELLED: Despite the identification of horseshoe bats as the reservoir of severe acute respiratory syndrome (SARS)-related coronaviruses (SARSr-CoVs), the origin of SARS-CoV ORF8, which contains the 29-nucleotide signature deletion among human strains, remains obscure. Although two SARS-related Rhinolophus sinicus bat CoVs (SARSr-Rs-BatCoVs) previously detected in Chinese horseshoe bats (Rhinolophus sinicus) in Yunnan, RsSHC014 and Rs3367, possessed 95% genome identities to human and civet SARSr-CoVs, their ORF8 protein exhibited only 32.2 to 33% amino acid identities to that of human/civet SARSr-CoVs. To elucidate the origin of SARS-CoV ORF8, we sampled 348 bats of various species in Yunnan, among which diverse alphacoronaviruses and betacoronaviruses, including potentially novel CoVs, were identified, with some showing potential interspecies transmission. The genomes of two betacoronaviruses, SARSr-Rf-BatCoV YNLF_31C and YNLF_34C, from greater horseshoe bats (Rhinolophus ferrumequinum), possessed 93% nucleotide identities to human/civet SARSr-CoV genomes. Although these two betacoronaviruses displayed lower similarities than SARSr-Rs-BatCoV RsSHC014 and Rs3367 in S protein to civet SARSr-CoVs, their ORF8 proteins demonstrated exceptionally high (80.4 to 81.3%) amino acid identities to that of human/civet SARSr-CoVs, compared to SARSr-BatCoVs from other horseshoe bats (23.2 to 37.3%). Potential recombination events were identified around ORF8 between SARSr-Rf-BatCoVs and SARSr-Rs-BatCoVs, leading to the generation of civet SARSr-CoVs. The expression of ORF8 subgenomic mRNA suggested that the ORF8 protein may be functional in SARSr-Rf-BatCoVs. The high Ka/Ks ratio among human SARS-CoVs compared to that among SARSr-BatCoVs supported that ORF8 is under strong positive selection during animal-to-human transmission. Molecular clock analysis using ORF1ab showed that SARSr-Rf-BatCoV YNLF_31C and YNLF_34C diverged from civet/human SARSr-CoVs in approximately 1990. SARS-CoV ORF8 originated from SARSr-CoVs of greater horseshoe bats through recombination, which may be important for animal-to-human transmission. IMPORTANCE: Although horseshoe bats are the primary reservoir of SARS-related coronaviruses (SARSr-CoVs), it is still unclear how these bat viruses have evolved to cross the species barrier to infect civets and humans. Most human SARS-CoV epidemic strains contain a signature 29-nucleotide deletion in ORF8, compared to civet SARSr-CoVs, suggesting that ORF8 may be important for interspecies transmission. However, the origin of SARS-CoV ORF8 remains obscure. In particular, SARSr-Rs-BatCoVs from Chinese horseshoe bats (Rhinolophus sinicus) exhibited <40% amino acid identities to human/civet SARS-CoV in the ORF8 protein. We detected diverse alphacoronaviruses and betacoronaviruses among various bat species in Yunnan, China, including two SARSr-Rf-BatCoVs from greater horseshoe bats that possessed ORF8 proteins with exceptionally high amino acid identities to that of human/civet SARSr-CoVs. We demonstrated recombination events around ORF8 between SARSr-Rf-BatCoVs and SARSr-Rs-BatCoVs, leading to the generation of civet SARSr-CoVs. Our findings offer insight into the evolutionary origin of SARS-CoV ORF8 protein, which was likely acquired from SARSr-CoVs of greater horseshoe bats through recombination.
Subject(s)
Coronavirus Infections/veterinary , Genome, Viral , RNA, Viral/genetics , Recombination, Genetic , Severe acute respiratory syndrome-related coronavirus/genetics , Viral Matrix Proteins/genetics , Amino Acid Sequence , Animals , Base Sequence , China , Chiroptera/virology , Coronavirus Infections/genetics , Coronavirus Infections/transmission , Coronavirus Infections/virology , Evolution, Molecular , Gene Expression , Humans , Molecular Sequence Data , Phylogeny , Phylogeography , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Viral/metabolism , Severe acute respiratory syndrome-related coronavirus/classification , Severe acute respiratory syndrome-related coronavirus/metabolism , Sequence Alignment , Sequence Homology, Nucleic Acid , Severe Acute Respiratory Syndrome/genetics , Severe Acute Respiratory Syndrome/metabolism , Severe Acute Respiratory Syndrome/transmission , Severe Acute Respiratory Syndrome/virology , Viral Matrix Proteins/metabolism , Viverridae/virologyABSTRACT
BACKGROUND: Penicillium marneffei is the most important thermal dimorphic fungus causing systemic mycosis in HIV-infected and other immunocompromised patients in Southeast Asia. However, laboratory diagnosis of penicilliosis, which relies on microscopic morphology and mycelial-to-yeast conversion, is time-consuming and expertise-dependent, thus delaying diagnosis and treatment. Although matrix -assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is useful for identification of various medically important fungi, its performance for identification of P. marneffei is less clear. RESULTS: We evaluated the performance of the Bruker MALDI-TOF MS system for identification of mold and yeast cultures of 59 clinical strains and the type strain of P. marneffei using the direct transfer method, with results compared to four phylogenetically closely related species, P. brevi-compactum, P. chrysogenum, Talaromyces aurantiacus and T. stipitatus. Using the Bruker original database combined with BDAL v4.0.0.1 and Filamentous Fungi Library 1.0, MALDI-TOF MS failed to identify the 60 P. marneffei strains grown in mold and yeast phase (identified as P. funiculosum and P. purpurogenum with scores <1.7 respectively). However, when the combined database was expanded with inclusion of spectra from 21 P. marneffei strains in mold and/or yeast phase, all the remaining 39 P. marneffei strains grown in mold or phase were correctly identified to the species level with score >2.0. The MS spectra of P. marneffei exhibited significant difference to those of P. brevi-compactum, P. chrysogenum, T. aurantiacus and T. stipitatus. However, MALDI-TOF MS failed to identify these four fungi to the species level using the combined database with or without spectra from P. marneffei. CONCLUSIONS: MALDI-TOF MS is useful for rapid identification of both yeast and mold cultures of P. marneffei and differentiation from related species. However, accurate identification to the species level requires database expansion using P. marneffei strains.
Subject(s)
Mycological Typing Techniques/methods , Mycoses/microbiology , Penicillium/chemistry , Penicillium/isolation & purification , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Fungi/chemistry , Fungi/classification , Fungi/isolation & purification , HumansABSTRACT
To identify potential biomarkers for improving diagnosis of melioidosis, we compared plasma metabolome profiles of melioidosis patients compared to patients with other bacteremia and controls without active infection, using ultra-high-performance liquid chromatography-electrospray ionization-quadruple time-of-flight mass spectrometry. Principal component analysis (PCA) showed that the metabolomic profiles of melioidosis patients are distinguishable from bacteremia patients and controls. Using multivariate and univariate analysis, 12 significant metabolites from four lipid classes, acylcarnitine (n = 6), lysophosphatidylethanolamine (LysoPE) (n = 3), sphingomyelins (SM) (n = 2) and phosphatidylcholine (PC) (n = 1), with significantly higher levels in melioidosis patients than bacteremia patients and controls, were identified. Ten of the 12 metabolites showed area-under-receiver operating characteristic curve (AUC) >0.80 when compared both between melioidosis and bacteremia patients, and between melioidosis patients and controls. SM(d18:2/16:0) possessed the largest AUC when compared, both between melioidosis and bacteremia patients (AUC 0.998, sensitivity 100% and specificity 91.7%), and between melioidosis patients and controls (AUC 1.000, sensitivity 96.7% and specificity 100%). Our results indicate that metabolome profiling might serve as a promising approach for diagnosis of melioidosis using patient plasma, with SM(d18:2/16:0) representing a potential biomarker. Since the 12 metabolites were related to various pathways for energy and lipid metabolism, further studies may reveal their possible role in the pathogenesis and host response in melioidosis.
Subject(s)
Melioidosis/blood , Metabolome , Sphingomyelins/blood , Bacteremia/blood , Biomarkers/blood , Carnitine/analogs & derivatives , Carnitine/blood , Case-Control Studies , Humans , Phosphatidylcholines/bloodABSTRACT
Although tuberculosis (TB) is a reemerging disease that affects people in developing countries and immunocompromised populations in developed countries, the current diagnostic methods are far from optimal. Metabolomics is increasingly being used for studies on infectious diseases. We performed metabolome profiling of plasma samples to identify potential biomarkers for diagnosing TB. We compared the plasma metabolome profiles of TB patients (n = 46) with those of community-acquired pneumonia (CAP) patients (n = 30) and controls without active infection (n = 30) using ultrahigh-performance liquid chromatography-electrospray ionization-quadrupole time of flight mass spectrometry (UHPLC-ESI-QTOFMS). Using multivariate and univariate analyses, four metabolites, 12R-hydroxy-5Z,8Z,10E,14Z-eicosatetraenoic acid [12(R)-HETE], ceramide (d18:1/16:0), cholesterol sulfate, and 4α-formyl-4ß-methyl-5α-cholesta-8-en-3ß-ol, were identified and found to have significantly higher levels in TB patients than those in CAP patients and controls. In a comparison of TB patients and controls, the four metabolites demonstrated area under the receiver operating characteristic curve (AUC) values of 0.914, 0.912, 0.905, and 0.856, sensitivities of 84.8%, 84.8%, 87.0%, and 89.1%, specificities of 90.0%, 86.7%, 86.7%, and 80.0%, and fold changes of 4.19, 26.15, 6.09, and 1.83, respectively. In a comparison of TB and CAP patients, the four metabolites demonstrated AUC values of 0.793, 0.717, 0.802, and 0.894, sensitivities of 89.1%, 71.7%, 80.4%, and 84.8%, specificities of 63.3%, 66.7%, 70.0%, and 83.3%, and fold changes of 4.69, 3.82, 3.75, and 2.16, respectively. 4α-Formyl-4ß-methyl-5α-cholesta-8-en-3ß-ol combined with 12(R)-HETE or cholesterol sulfate offered ≥70% sensitivity and ≥90% specificity for differentiating TB patients from controls or CAP patients. These novel plasma biomarkers, especially 12(R)-HETE and 4α-formyl-4ß-methyl-5α-cholesta-8-en-3ß-ol, alone or in combination, are potentially useful for rapid and noninvasive diagnosis of TB. The present findings may offer insights into the pathogenesis and host response in TB.
Subject(s)
Biomarkers/blood , Metabolome , Plasma/chemistry , Tuberculosis/diagnosis , Tuberculosis/pathology , Adolescent , Adult , Aged , Aged, 80 and over , Chromatography, Liquid , Female , Humans , Male , Mass Spectrometry/methods , Middle Aged , ROC Curve , Sensitivity and Specificity , Young AdultABSTRACT
Penicillium marneffei is an opportunistic fungal pathogen endemic in Southeast Asia, causing lethal systemic infections in immunocompromised patients. P. marneffei grows in a mycelial form at the ambient temperature of 25°C and transitions to a yeast form at 37°C. The ability to alternate between the mycelial and yeast forms at different temperatures, namely, thermal dimorphism, has long been considered critical for the pathogenicity of P. marneffei, yet the underlying genetic mechanisms remain elusive. Here we employed high-throughput sequencing to unravel global transcriptional profiles of P. marneffei PM1 grown at 25 and 37°C. Among â¼11,000 protein-coding genes, 1,447 were overexpressed and 1,414 were underexpressed at 37°C. Counterintuitively, heat-responsive genes, predicted in P. marneffei through sequence comparison, did not tend to be overexpressed at 37°C. These results suggest that P. marneffei may take a distinct strategy of genetic regulation at the elevated temperature; the current knowledge concerning fungal heat response, based on studies of model fungal organisms, may not be applicable to P. marneffei. Our results further showed that the tandem repeat sequences (TRSs) are overrepresented in coding regions of P. marneffei genes, and TRS-containing genes tend to be overexpressed at 37°C. Furthermore, genomic sequences and expression data were integrated to characterize gene clusters, multigene families, and species-specific genes of P. marneffei. In sum, we present an integrated analysis and a comprehensive resource toward a better understanding of temperature-dependent genetic regulation in P. marneffei.
Subject(s)
Gene Expression Regulation, Fungal , Heat-Shock Response/genetics , Penicillium/metabolism , Transcription, Genetic , Genes, Fungal , Heat-Shock Proteins/genetics , Hot Temperature , Penicillium/genetics , Tandem Repeat Sequences , TranscriptomeABSTRACT
BACKGROUND: Patients with developmental dysplasia of the hip (DDH) are at risk for residual acetabular dysplasia even after successful closed reduction. The aim of this study was to identify predictors of long-term outcomes in order to guide prognostication and management. METHODS: Patients who were treated for DDH at 2 institutions between 1970 and 2010 and had follow-up until skeletal maturity were screened for inclusion. Hips that underwent open reduction were excluded to reduce iatrogenic confounding. Syndromal (including neuromuscular and arthrogrypotic) hip instability with collagenopathies were excluded. Demographic characteristics, Tönnis grade, age at the time of the reduction, surgical treatment, acetabular index, lateral center-edge angle, residual dysplasia graded using the Severin classification, and the presence and type of proximal femoral growth disturbance categorized using the Bucholz and Ogden classification were assessed. In addition, the the acetabular angle was recorded at the latest follow-up before secondary procedures, and the Oxford Hip Score and 5-level EuroQoL (EQ)-5 Dimension score were recorded at the latest follow-up. To account for repeated measures, generalized estimating equations (GEE) logistic regression was utilized for the multivariable analysis. A support vector machine model and a receiver operating characteristic curve analysis were utilized to identify prognostication thresholds. RESULTS: One hundred and seven hips (96 female, 11 male) that were followed to skeletal maturity, with a mean follow-up of 20 years (range, 10 to 54 years), were included in the analysis. Eighty-nine hips (83%) demonstrated a good outcome at skeletal maturity, with a Severin grade of I or II. Major growth disturbances of Bucholz and Ogden types II, III, or IV were present in 13 hips (12%). At the latest follow-up after skeletal maturity (before any secondary procedures), the mean acetabular angle was 45° ± 4° and the mean lateral center-edge angle was 26° ± 8°. The mean Oxford Hip Score and EQ visual analog scale values were 47 and 86, respectively. A GEE logistic regression analysis of 1,135 observations revealed that the acetabular index (odds ratio [OR], 1.16 per degree; p < 0.001) and age (OR, 1.20 per year; p = 0.003) were significant predictors of a poor outcome (i.e., Severin grade III, IV, or V). Significant differences in acetabular indices across all age groups were found between hips with a good outcome and those with a poor outcome. Age-specific acetabular index prognostication cutoff values are presented. CONCLUSIONS: This long-term follow-up study demonstrated that the age-specific acetabular index remains an important predictor of residual dysplasia at skeletal maturity. The proposed prognostication chart and thresholds herein can help to guide orthopaedic surgeons and parents when contemplating the use of an intervention versus surveillance to optimize long-term outcomes. LEVEL OF EVIDENCE: Prognostic Level III. See Instructions for Authors for a complete description of levels of evidence.
ABSTRACT
Penicillium marneffei is the most important thermal dimorphic, pathogenic fungus endemic in China and Southeast Asia and is particularly important in HIV-positive patients. We report the 28,887,485-bp draft genome sequence of P. marneffei, which contains its complete mitochondrial genome, sexual cycle genes, a high diversity of Mp1p homologues, and polyketide synthase genes.
Subject(s)
Genome, Fungal , Penicillium/genetics , Base Sequence , Molecular Sequence Data , Sequence Analysis, DNAABSTRACT
Human oviductal cells produce complement-3 (C3) and its derivative, iC3b. These molecules are important in immune responses. Our recent study suggested that iC3b also possessed embryotrophic activity and it stimulates the blastulation and hatching rates of in vitro cultured mouse embryos. The objective is to study the impact of C3 deficiency on early pregnancy in vivo using homozygous C3-deficient (C3KO) and wild-type (C3WT) mice. C3 protein was undetectable in the reproductive tissues of C3KO mice. Deficiency in C3 is associated with significantly longer estrous cycle (P = 0.037). No significant difference was found in the ovulation rate, total cell count in blastocysts and implantation rate between the wild-type and the C3KO mice, though C3KO mice tended to have lower values in the latter two parameters. On day 15 of pregnancy, C3KO mice had fewer conceptus (P < 0.001) and higher resorption rate (P < 0.001) than that of C3WT mice. The fetal and placental weights (P < 0.001) were lower in the C3KO mice. The placenta of C3KO mice had smaller spongiotrophoblast (P = 0.001) and labyrinth (P = 0.037). Deficiency in C3 is associated with mild impairment in early pregnancy including longer estrous cycle and higher resorption rates after implantation. The impairment may be related to compromised placental development leading to under-developed fetuses.
Subject(s)
Complement C3/deficiency , Embryo Implantation , Embryo Loss , Animals , Blastocyst/cytology , Complement C3/genetics , Complement C3/metabolism , Estrous Cycle , Female , Fetal Viability , Fetal Weight , Gene Expression , Genitalia, Female/metabolism , Male , Mice , Mice, Knockout , Ovulation , Placenta/anatomy & histology , Placenta/physiology , Polymerase Chain Reaction , PregnancyABSTRACT
The human oviduct derived embryotrophic factor-3 (ETF-3) contains complement protein-3 (C3) and its derivates. Although C3 is not embryotrophic, it is converted into the embryotrophic derivative, iC3b in the presence of embryos and oviductal cells. The regulation of C3 production in the oviduct is not known. The objectives of this study were to investigate the effects of presence of preimplantation embryos and hormones on C3 expression in the oviducts in vitro and in vivo. The expression of C3 in the oviduct of pregnant mice was compared to that of pseudo-pregnant mice. The hormonal action on C3 expression was studied in the ovariectomized mouse oviducts and human oviductal epithelial (OE) cells. The results showed that the level of C3 mRNA in the mouse oviduct was high on Day 1 and Day 2, but decreased to a minimum on Day 4 of pregnancy, whereas that of pseudo-pregnancy remained relatively stable within the same period. The protein levels of C3 and iC3b specific fragments, alpha-115 and alpha-40, respectively in the mouse oviductal luminal fluid were highest on Day 3 of pregnancy, when the embryos were expected to be most sensitive to the embryotrophic activity of ETF-3. Estrogen elevated C3 expression in the ovariectomized mouse oviduct and the OE cells. Progesterone suppressed estrogen-induced C3 expression in the mouse oviduct, but had no effect on OE cells. In conclusion, the presence of embryo and steroid hormones regulate the synthesis and secretion of oviductal C3.
Subject(s)
Complement C3/metabolism , Fallopian Tubes/metabolism , Gene Expression Regulation/physiology , Analysis of Variance , Animals , Cell Line , Complement C3/genetics , Complement C3b/metabolism , Estrogens/blood , Estrogens/metabolism , Estrogens/pharmacology , Fallopian Tubes/cytology , Female , Gene Expression Regulation/drug effects , Humans , Male , Mice , Ovariectomy , Pregnancy , Progesterone/blood , Progesterone/metabolism , Progesterone/pharmacology , Pseudopregnancy/metabolismABSTRACT
BACKGROUND: Atypical triplane fractures are defined as triplane fractures that are intra-articular but affect the non-weight-bearing area of the tibia plafond or extra-articular triplane fractures where the epiphyseal fracture line exits outside the articulating cortex of the medial malleolus. These fractures are scarcely reported in the literature. Here, we study the fracture pattern, mechanisms, and recommendations for management. METHODS: This is a retrospective study of all triplane fractures identified from 2012 to 2016 in a tertiary referral center. There were 10 atypical triplane fracture patterns identified in this cohort. All patients were followed up with an average of 19 months. A modified atypical triplane fracture classification was devised and compared with previously reported classification systems. Clinical outcomes measured included treatment complications, ankle range of motion, and time needed to return to sports. RESULTS: We identified a new extra-articular triplane fracture variant with an anteromedial epiphyseal sleeve fragment (fracture variant). There were no long-term complications from operative closed reduction and percutaneous screw fixation. Operative cases had earlier ankle mobilization and regained full range of motion (12.8 weeks vs 13.3 weeks) earlier. The average time to return to sports was 5.2 months. CONCLUSIONS: We propose a modified classification for atypical triplane fractures and recommend closed reduction and percutaneous screw fixation for displaced atypical triplane fractures. LEVEL OF EVIDENCE: IV (Case Series).
Subject(s)
Intra-Articular Fractures/classification , Tibial Fractures/classification , Adolescent , Ankle Joint , Child , Early Ambulation , Epiphyses , Female , Fracture Fixation, Internal , Humans , Intra-Articular Fractures/diagnostic imaging , Intra-Articular Fractures/surgery , Male , Range of Motion, Articular , Retrospective Studies , Tibial Fractures/diagnostic imaging , Tibial Fractures/surgery , Tomography, X-Ray ComputedABSTRACT
OBJECTIVE: The aim of the study is to investigate the long-term outcome of patients who received Lambrinudi arthrodesis for severe equinovarus deformities. METHODS: This is a single-center, retrospective study of patients who received Lambrinudi triple arthrodesis of the foot. Both clinical and radiological information were analyzed. RESULTs: We were able to review 10 patients suffering from severe equinus deformities mostly as a result of or in association with poliomyelitis (8 of 10) who received Lambrinudi arthrodesis. The majority (7 of 10) of our patients had fair to good outcome at an average follow-up of 37 years. Specifically, six of eight polio patients had fair to good outcome. Of the X-rays available for assessment, the majority of patients showed radiological signs of adjacent joint arthritis; however, the presence of such did not invariably lead to poor clinical outcome. CONCLUSION: Lambrinudi arthrodesis is a treatment option with favorable long-term outcome for patients with severe, fixed equinus deformities.
Subject(s)
Arthrodesis , Clubfoot/surgery , Adolescent , Adult , Arthritis , Clubfoot/diagnostic imaging , Clubfoot/etiology , Female , Follow-Up Studies , Humans , Male , Middle Aged , Poliomyelitis/complications , Radiography , Retrospective Studies , Time Factors , Treatment Outcome , Young AdultABSTRACT
Human oviductal epithelial (OE) cells produce complement protein 3 (C3) and its derivatives, C3b and inactivated complement-3b (iC3b). Among them, iC3b is the most potent embryotrophic molecule. We studied the production of iC3b in the oviductal cell/embryo culture system. In the immune system, C3 convertase converts C3 into C3b, and the conversion of C3b to iC3b requires factor I (fI) and its cofactors, such as factor H or membrane cofactor protein. Human oviductal epithelium and OE cells expressed mRNA and protein of the components of C3 convertase, including C2, C4, factor B, and factor D. The OE cell-conditioned medium contained active C3 convertase activity that was suppressed by C3 convertase inhibitor, H17 in a dose and time-dependent manner. Although the oviductal epithelium and OE cells produced fI, the production of its cofactor, factor H required for the conversion of C3b to iC3b, was weak. Thus, OE cell-conditioned medium was inefficient in producing iC3b from exogenous C3b. On the contrary, mouse embryos facilitated such conversion to iC3b, which was taken up by the embryos, resulting in the formation of more blastocysts of larger size. The facilitatory activity was mediated by complement receptor 1-related gene/protein Y (Crry) with known membrane cofactor protein activity on the trophectoderm of the embryos as anti-Crry antibody inhibited the conversion and embryotrophic activity of C3b in the presence of fI. In conclusion, human oviduct possesses C3 convertase activity converting C3 to C3b, and Crry of the preimplantation embryos may be involved in the production of embryotrophic iC3b on the surface of the embryos.
Subject(s)
Blastocyst/metabolism , Cell Communication/physiology , Complement C3b/metabolism , Fallopian Tubes/metabolism , Fallopian Tubes/pathology , Animals , Cells, Cultured , Complement C3-C5 Convertases/metabolism , Complement Factor D/metabolism , Complement Factor H/metabolism , Complement Factor I/metabolism , Female , Humans , Mice , Mice, Inbred ICR , Receptors, Complement/metabolism , Receptors, Complement 3bABSTRACT
Paediatric pelvic and hip radiographs are a common investigation used when assessing a child for suspected developmental dysplasia of the hip. This report describes an attempt to establish normal values of medial joint space, acetabular index and centre edge angle according to specific age groups and sex in a Chinese population. Patients who had undergone a pelvic radiograph as part of their assessment, but were subsequently found to have normal hips were recruited retrospectively. These patients were grouped according to sex and age; medial joint space, acetabular index and centre edge angle were measured in all radiographs. A mean±SD was calculated for each group, and then each age group was tested for statistical significance between the male and the female groups. A total of, 98 patients were recruited, who underwent 188 pelvic radiographs, resulting in images of 376 'normal' hips. The results for medial joint space, acetabular index and centre edge angle for each age and sex group are described. Only the acetabular index requires different reference ranges for male and female patients because of consistent statistical significance between the two groups. It was found that medial joint space remained fairly constant throughout the age groups, whereas the acetabular index decreased and the centre edge angle increased slightly. The reference ranges for the parameters described here are quite different from those established previously in a population of Northern-European descent, which could be because of a variety of reasons including genetics, body habitus and measurement technique. We believe that it would be prudent to implement these different ranges when assessing patients of Chinese heritage to optimize care of patients who may suffer as a consequence of not receiving treatment for missed hip dysplasia. LEVEL OF EVIDENCE: Diagnostic Study Level III - Study of nonconsecutive patients (without consistently applying the reference 'gold' standard).
Subject(s)
Acetabulum/diagnostic imaging , Femur Head/diagnostic imaging , Hip Joint/diagnostic imaging , Acetabulum/anatomy & histology , Asian People , Biomechanical Phenomena , Child , Child, Preschool , China , Female , Femur Head/anatomy & histology , Hip Joint/anatomy & histology , Humans , Infant , Infant, Newborn , Male , Radiography , Reference Values , Retrospective StudiesABSTRACT
Despite the recent emergence of enterovirus D68 (EV-D68), its clinical impact on adult population is less well defined. To better define the epidemiology of EV-D68, 6,800 nasopharyngeal aspirates (NPAs) from 2010-2014 were subject to EV-D68 detection by RT-PCR and sequencing of 5'UTR and partial VP1. EV-D68 was detected in 30 (0.44%) NPAs from 22 children and 8 adults/elderlies. Sixteen patients (including five elderly) (53%) had pneumonia and 13 (43%) patients were complicated by small airway disease exacerbation. Phylogenetic analysis of VP1, 2C and 3D regions showed four distinct lineages of EV-D68, clade A1, A2, B1 and B3, with adults/elderlies exclusively infected by clade A2. The potentially new clade, B3, has emerged in 2014, while strains closely related to recently emerged B1 strains in the United States were also detected as early as 2011 in Hong Kong. The four lineages possessed distinct aa sequence patterns in BC and DE loops. Amino acid residues 97 and 140, within BC and DE-surface loops of VP1 respectively, were under potential positive selection. EV-D68 infections in Hong Kong usually peak in spring/summer, though with a delayed autumn/winter peak in 2011. This report suggests that EV-D68 may cause severe respiratory illness in adults/elderlies with underlying co-morbidities.
Subject(s)
Enterovirus D, Human/classification , Enterovirus Infections/virology , Respiratory Tract Infections/virology , Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Enterovirus D, Human/genetics , Enterovirus Infections/epidemiology , Female , Hong Kong/epidemiology , Humans , Infant , Male , Middle Aged , Phylogeny , Respiratory Tract Infections/epidemiology , Seasons , Sequence Analysis, RNA , United StatesABSTRACT
Unlike Elizabethkingia meningoseptica, the clinical importance of E. anophelis is poorly understood. We determined the clinical and molecular epidemiology of bacteremia caused by Elizabethkingia-like species from five regional hospitals in Hong Kong. Among 45 episodes of Elizabethkingia-like bacteremia, 21 were caused by Elizabethkingia, including 17 E. anophelis, three E. meningoseptica and one E. miricola; while 24 were caused by other diverse genera/species, as determined by 16S rRNA gene sequencing. Of the 17 cases of E. anophelis bacteremia, 15 (88%) were clinically significant. The most common diagnosis was pneumonia (n = 5), followed by catheter-related bacteremia (n = 4), neonatal meningitis (n = 3), nosocomial bacteremia (n = 2) and neutropenic fever (n = 1). E. anophelis bacteremia was commonly associated with complications and carried 23.5% mortality. In contrast, of the 24 episodes of bacteremia due to non-Elizabethkingia species, 16 (67%) were clinically insignificant. Compared to non-Elizabethkingia bacteremia, Elizabethkingia bacteremia was associated with more clinically significant infections (P < 0.01) and positive cultures from other sites (P < 0.01), less polymicrobial bacteremia (P < 0.01), and higher complication (P < 0.05) and mortality (P < 0.05) rates. Elizabethkingia bacteremia is predominantly caused by E. anophelis instead of E. meningoseptica. Elizabethkingia bacteremia, especially due to E. anophelis, carries significant morbidity and mortality, and should be considered clinically significant unless proven otherwise.