The microbiological safety of reconstituted infant formula (RIF) has focused on infectious pathogens, whereas the risk of spore-forming bacteria (SFB) has been limited to spoilage and toxin production. This study suggests an underrecognized niche of SFB as nitrite producers during the handling of RIF. The production of nitrite along with the bacterial growth of 133 nitrite-producing SFB isolated from infant formula processing environments and end-products (70 mesophiles and 63 thermophiles) under RIF handling conditions were analysed. Most mesophiles (68 out of 70) and two thermophiles showed nitrite production during growth at 30 degrees C or 40 degrees C. Vigorous producers of nitrite [Bacillus sp. strains (FHS-PPBM449, 481, 236, 237)] showed a rapid onset of nitrite production (within 4 h). In particular, FHS-PPBM449 (2-3 log CFU/mL) exhibited the shortest onset time (210 min) and a nitrite production level up to 521 mu M in RIF with 100 ppm nitrate at 40 degrees C. Overall, the results of the maximum level of nitrite produced by vigorous nitrite producers indicate that infants can consume more than seven times the acceptable daily intake of nitrite (0.74 mg for 12-month-old infants with an average body weight), even via a single feeding of RIF. An analysis of the relationship of the onset time of nitrite production with the bacterial concentration based on predictive models suggests that the growth of SFB up to 5-6 log CFU/ mL is regarded as a prerequisite for nitrite production. This study revealed an underreported source of nitrite from RIF handling conditions, and the rapid onset of a high level of nitrite production from SFB should be the major target in the establishment of intervention strategies against nitrite as a microbial risk.

Salted napa cabbage is the most important ingredient of kimchi. Currently, people have started to prepare ready-to-use salted napa cabbage at home. This study focused on this trend by investigating consumers' beliefs, opinions, and actual use of the products by conducting a telephone survey (895 female consumers) and face-to-face interviews (n 1/4 514) in 2016 and a telephone survey (n 1/4 200) in 2021. Most respondents (93 and 91% in 2016 and 2021, respectively) answered that convenience was the main reason for using salted napa cabbage. Regarding consumption behavior, 22 and 16% of the respondents in each year used salted napa cabbage after storing it for more than 24 h. In particular, 85 and 91% of consumers stored the product at room temperature, and 60 and 58% used it without washing, which could affect the quality of the food, as microorganisms could multiply during the storage. Inappropriate handling increased by age group, especially in 2021 (P, 0.05). In the query on satisfaction after using the products, 85 and 80% of respondents were satisfied because the product was convenient (54%) and hygienic (17%). Conversely, the respondents who were not satisfied with the products did not like the degree of salting and unhygienic status of the products. The majority (93 and 80%) of consumers preferred buying salted napa cabbage again because of its convenience. Although consumers thought that hygiene and quality were important factors, many respondents (83 and 72%) were not aware of foodborne illnesses associated with kimchi. Nevertheless, consumers intended to pay more for safe, salted napa cabbage (72 and 76%). The results of this study provide useful and credible data for understanding the factors affecting consumers' consumption and general beliefs and opinions on the use of salted napa cabbage, especially for food safety management.

Fibrodysplasia ossificans progressiva (FOP) is a rare and devastating genetic disease, in which soft connective tissue is converted into heterotopic bone through an endochondral ossification process. Patients succumb early as they gradually become trapped in a second skeleton of heterotopic bone. Although the underlying genetic defect is long known, the inherent complexity of the disease has hindered the discovery of effective preventions and treatments. New developments in the gene therapy field have motivated its consideration as an attractive therapeutic option for FOP. However, the immune system's role in FOP activation and the as-yet unknown primary causative cell, are crucial issues which must be taken into account in the therapy design. While gene therapy offers a potential therapeutic solution, more knowledge about FOP is needed to enable its optimal and safe application.

An 11-year-old Korean boy presented with short stature, hip dysplasia, radial head dislocation, carpal coalition, genu valgum, and fixed patellar dislocation and was clinically diagnosed with Steel syndrome. Scrutinizing the trio whole-exome sequencing data revealed novel compound heterozygous mutations of COL27A1 (c.[4229_4233dup]; [3718_5436del], p.[Gly1412Argfs*157];[Gly1240_Lys1812del]) in the proband, which were inherited from heterozygous parents. The maternal mutation was a large deletion encompassing exons 38-60, which was challenging to detect.

Purpose The purpose of this study was to identify imaging risk factors on contrast-enhanced hip MRI after closed reduction of developmental dysplasia of the hip (DDH) that could predict future development of avascular necrosis (AVN) of the femoral head. Materials and methods Fifty-eight infants (F: M = 53: 5, aged 3-18 months) who underwent immediate postoperative contrast-enhanced hip MRI after closed reduction of DDH were included in this study. Quality of reduction (concentric vs eccentric reduction with or without obstacles), abduction angle of the hip, presence of ossific nucleus, and pattern of contrast enhancement of the femoral head were retrospectively evaluated on MRI. Interobserver agreement of contrast enhancement pattern on MRI were evaluated by two radiologists. Development of AVN was determined through radiographic findings at 1 year after reduction. Results AVN of the femoral head developed in 13 (22%) of 58 patients. Excessive abduction of the hip joint (OR 4.65, [95% CI 1.20, 18.06] and global decreased enhancement of the femoral head (OR 71.66, [95% CI 10.54, 487.31]) exhibited statistically significant differences between the AVN and non-AVN groups (P < 0.05). Eccentric reduction (P = 0.320) did not show statistically significant difference between two groups and invisible ossific nucleus (P = 0.05) showed borderline significance. Multi-variable logistic regression indicated that global decreased enhancement of the femoral head was a significant risk factor of AVN (OR 27.92, 95% CI [4.17, 350.18]) (P = 0.0031). Interobserver agreement of contrast enhancement pattern analysis and diagnosis of AVN were good (0.66, 95% CI [0.52, 0.80]). Conclusion Contrast-enhanced hip MRI provides accurate anatomical assessment of the hip after closed reduction of DDH. Global decreased enhancement of the femoral head could be used as a good predictor for future development of AVN after closed reduction of DDH.

Spondyloepimetaphyseal dysplasias (SEMDs) are a heterogeneous group of disorders with variable growth failure and skeletal impairments affecting the spine and long bone epiphyses and metaphyses. Here we report on four unrelated families with SEMD in which we identified two monoallelic missense variants and one monoallelic splice site variant inRPL13, encoding the ribosomal protein eL13. In two out of four families, we observed autosomal dominant inheritance with incomplete penetrance and variable clinical expressivity; the phenotypes of the mutation-positive subjects ranged from normal height with or without hip dysplasia to severe SEMD with severe short stature and marked skeletal dysplasia.In vitrostudies on patient-derived dermal fibroblasts harboringRPL13missense mutations demonstrated normal eL13 expression, with proper subcellular localization but reduced colocalization with eL28 (p< 0.001). Cellular functional defects in fibroblasts from mutation-positive subjects indicated a significant increase in the ratio of 60S subunits to 80S ribosomes (p= 0.007) and attenuated global translation (p= 0.017). In line with the human phenotype, ourrpl13mutant zebrafish model, generated by CRISPR-Cas9 editing, showed cartilage deformities at embryonic and juvenile stages. These findings extend the genetic spectrum ofRPL13mutations causing this novel human ribosomopathy with variable skeletal features. Our study underscores for the first time incomplete penetrance and broad phenotypic variability in SEMD-RPL13 type and confirms impaired ribosomal function. Furthermore, the newly generatedrpl13mutant zebrafish model corroborates the role of eL13 in skeletogenesis. (c) 2020 The Authors.Journal of Bone and Mineral Researchpublished by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR)..

BACKGROUND:The GNAS gene on chromosome 20q13.3 is a complex, imprinted locus regulated in a tissue-specific manner. GNAS inactivation disorders are a heterogeneous group of rare disorders caused by mutations and methylation defects. These are divided into pseudohypoparathyroidism (PHP) types 1A and 1B, pseudo-pseudohypoparathyroidism (PPHP), and progressive osseous heteroplasia (POH), depending on the presence or absence of hormone resistance, Albright's hereditary osteodystrophy (AHO), and ectopic ossification.METHODS:This study analyzed the clinical characteristics and molecular genetic backgrounds of 18 Korean patients from 16 families with a genetically confirmed GNAS defect. Auxological parameters, AHO phenotypes, types of hormonal resistance, family history, and molecular genetic disturbances were reviewed retrospectively.RESULTS:Nine (90%) patients with PHP1A showed resistance to parathyroid hormone (PTH) and all patients showed elevated thyroid-stimulating hormone (TSH) levels at diagnosis. Eight (80%) patients were managed with levothyroxine supplementation. Three of six patients with PHP1B had elevated TSH levels, but none of whom needed levothyroxine medication. AHO features were absent in PHP1B. Patients with PPHP and POH did not show any hormone resistance, and both of them were born as small for gestational age. Among the 11 families with PHP1A, PPHP, and POH, eight different (three novel) mutations in the GNAS gene were identified. Among the six patients with PHP1B, two were sporadic cases and four showed isolated loss of methylation at GNAS A/B:TSS-DMR.CONCLUSIONS:Clinical and molecular characteristics of Korean patients with GNAS inactivation disorders were described in this study. Also, we reaffirmed heterogeneity of PHP, contributing to further accumulation and expansion of current knowledge of this complex disease.

Background A high fibular location on plain radiographs has been considered suggestive of a discoid lateral meniscus (DLM). We sought to determine whether the high location of the fibular head observed in DLM patients represents a true pathologic finding. Methods Eighty-two patients with symptomatic DLM who underwent arthroscopic treatment constituted the study group (DLM group). The same number of age-matched patients without DLM (control group) were recruited. The DLM group was further divided according to the presence or absence of a knee extension block (EB) of more than 10 degrees into EB(+) and EB(-) subgroups. We defined the compensated distance to the fibular head (CDF) as the distance from the lateral joint line to the tip of the fibular head divided by the femoral condylar width. CDF values measured on anteroposterior (AP) plain radiographs and sagittal MR images were compared between the DLM and control groups and between the EB(+) and EB(-) subgroups. Results There was no significant difference in the mean preoperative CDF value on either AP plain radiographs or sagittal MR images between the DLM and control groups. However, the mean CDF value on AP plain radiographs was significantly lower in the EB(+) subgroup than in the EB(-) subgroup ( p =0.011). The knee EB disappeared in all patients after arthroscopic treatment when the CDF on AP plain radiographs showed no significant difference between subgroups ( p >0.05). Linear regression analysis revealed that the CDF on AP plain radiographs decreased significantly when knee flexion was increased from 0 to 30 degrees ( p =0.003). Conclusions The high fibular location observed on AP plain radiography in DLM patients was associated with limited knee extension. Altered X-ray beam projection to the flexed knee creates the false appearance of a high fibular location on AP plain radiography.

Background and purpose — There are few studies on overgrowth of the affected limb after treatment of developmental dysplasia of the hip (DDH). We investigated the incidence of overgrowth and its risk factors in DDH patients. Patients and methods — 101 patients were included in this study. Overgrowth was defined by 2 criteria: when the height of the femoral head of the affected side was higher than that of the contralateral side by more than 10 mm, or by more than 15 mm. The potential risk factors of distinct overgrowth were retrospectively examined using multivariable analysis. Results — When overgrowth was defined as femoral head height difference (FHHD) > 10 mm, its incidence was 44%, and only femoral osteotomy was identified as a significant risk factor with a relative risk (RR) of 1.6 (95% confidence interval [CI] 1.0–2.5). When overgrowth was defined as FHHD > 15 mm, its incidence was 23%, and femoral osteotomy was identified as the only significant risk factor with an RR of 2.3 (CI 1.2–4.5). Overgrowth developed more frequently in patients who underwent femoral osteotomy at the age of 2 to 4 years (87%) than in the others (46%) (p = 0.04). Interpretation — Overgrowth of the affected limb is common in DDH patients. Patients who underwent femoral osteotomy, especially at the age of 2 to 4 years, may require careful follow-up because of the substantial risk for overgrowth.

Liang, Zhen Chang MBBS, MRCS, DipSpMed, PhD, MBA; Chong, Mark Seng Ye BMBS, BMedSc, FRCS; Liu, Gabriel Ka Po MBBS, MSc, FRCS; Valle, Alejandro Gonzalez Della MD; Wang, Dahui MD, PhD; Lyu, Xuemin MD, PhD; Chang, Chih-Hung MD, PhD, FICS; Cho, Tae-Joon MD; Haas, Steven B. MD, MPH; Fisher, Dale MBBS, FRACP; Murphy, Diarmuid MBBS, FRCS; Hui, James Hoi Po MBBS, MD, FRCS Author Information

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Distal renal tubular acidosis (dRTA) is a rare renal tubular disorder characterized by normal anion gap metabolic acidosis, hypokalemia, and high urine pH. It can be inherited or acquired. In untreated pediatric patients with dRTA, rickets and growth retardation are common. We report the case of a 12-year-old Lao girl who presented with typical clinical features of dRTA with severe bone deformities that developed after a bed-ridden state due to a bicycle accident at the age of 8 years. Initial laboratory tests revealed metabolic acidosis with a normal anion gap, hypokalemia, and alkali urine. Renal ultrasonography revealed bilateral medullary nephrocalcinosis. Whole exome sequencing revealed no pathogenic mutations. After treatment with oral alkali, potassium, and vitamin D, she could walk and run. Later, she underwent corrective orthopedic surgeries for bony deformities. Thus, in pediatric dRTA patients, despite severe symptoms remaining untreated, accurate diagnosis and proper management can improve quality of life.

PURPOSE To examine the efficacy, safety, and clinical outcomes of distraction osteogenesis through the physis (PDO) or through subphyseal osteotomy (SPDO) in patients with atrophic-type congenital pseudarthrosis of tibia with proximal tibial dysplasia. METHODS To validate the efficacy and safety of PDO and SPDO, radiographic and clinical parameters were compared between 5 patients who underwent proximal tibial metaphyseal or metadiaphyseal lengthening as a control (group 1) and 7 patients who underwent PDO or SPDO (group 2). Postoperative complication was also compared between the groups. RESULTS A significant difference in terms of healing index (group 1, 83.3±24.7 d/cm; group 2, 35.0±11.1 d/cm; P=0.001) and percentage increase (11.0%±3.7% vs. 23.1%±10.5%, P=0.034) was observed between the 2 groups. According to the Paley classification, group 1 included 1 "problems" case and 3 "obstacles" cases, whereas group 2 included 2 "problems" cases and 1 "obstacles" case. According to the Lascombes classification, group 1 included 2 grade IIIb cases and 3 grade IV cases, whereas group 2 included 6 grade I cases and 1 grade IIa case. Severe complications were significantly higher in group 1 compared with the group 2 (P=0.007). CONCLUSIONS This study demonstrated that PDO or SPDO can be effectively and safely performed for tibial lengthening in atrophic-type congenital pseudarthrosis of tibia patients with proximal tibial dysplasia. LEVEL OF EVIDENCE Level III.

Because Mg-Ca-Zn alloys are biodegradable and obviate secondary implant removal, they are especially beneficial for pediatric patients. We examined the degradation performance of Mg-Ca-Zn alloys depending on the surface modification and investigated the in vivo effects on the growth plate in a skeletally immature rabbit model. Either plasma electrolyte oxidation (PEO)-coated (n = 18) or non-coated (n = 18) Mg-Ca-Zn alloy was inserted at the distal femoral physis. We measured the degradation performance and femoral segment lengths using micro-CT. In addition, we analyzed the histomorphometric and histopathologic characteristics of the growth plate. Although there were no acute, chronic inflammatory reactions in either group, they differed significantly in the tissue reactions to their degradation performance and physeal responses. Compared to non-coated alloys, PEO-coated alloys degraded significantly slowly with diminished hydrogen gas formation. Depending on the degradation rate, large bone bridge formation and premature physeal arrest occurred primarily in the non-coated group, whereas only a small-sized bone bridge formed in the PEO-coated group. This difference ultimately led to significant shortening of the femoral segment in the non-coated group. This study suggests that optimal degradation could be achieved with PEO-coated Mg-Ca-Zn alloys, making them promising and safe biodegradable materials with no growth plate damage.

PURPOSE:Skeletal dysplasias comprise a heterogeneous group of inherited disorders of development, growth, and maintenance of the human skeleton. Because of their relative rarity and wide phenotypic variability, patients should be accurately identified, uniformly assessed, and managed by clinicians who are aware of their potential complications and possess the knowledge and resources to treat them effectively. This study presents expert guidelines developed to improve the diagnosis and management of patients with type II collagen skeletal disorders to optimize clinical outcomes.METHODS:A panel of 11 multidisciplinary international experts in the field of skeletal dysplasia participated in a Delphi process, which comprised analysis of a thorough literature review with subsequent generation of 26 diagnosis and care recommendations, followed by two rounds of anonymous voting with an intervening face-to-face meeting. Those recommendations with more than 80% agreement were considered as consensual.RESULTS:After the first voting round, consensus was reached to support 12 of 26 (46%) statements. After the panel discussion, the group reached consensus on 22 of 24 revised statements (92%).CONCLUSIONS:Consensus-based, expert best practice guidelines developed as a standard of care to assist accurate diagnosis, minimize associated health risks, and improve clinical outcomes for patients with type II collagen skeletal dysplasias.

SPONASTRIME dysplasia is a rare recessive skeletal dysplasia characterized by short 50 stature, facial dysmorphism, and aberrant radiographic findings of the spine and long bone 51 metaphysis. No causative genetic alterations for SPONASTRIME dysplasia have yet been 52 determined. Using whole exome sequencing, we identified biallelic TONSL mutations in 10 of 13 53 individuals with SPONASTRIME dysplasia. TONSL is a multi-domain scaffold protein that interacts 54 with DNA replication and repair factors, and plays critical roles in resistance to replication stress and 55 maintenance of genome integrity. We show here that cellular defects in dermal fibroblasts from 56 individuals are complemented by expressing wild-type TONSL . In addition, in vitro cell-based assays 57 and in silico analyses of TONSL structure support the pathogenicity of those TONSL variants. 58 Intriguingly, a knock-in Tonsl mouse model leads to embryonic lethality, implying the physiological 59 importance of TONSL. Overall, these findings indicate that genetic variants resulting in reduced 60 function of TONSL cause SPONASTRIME dysplasia and highlight the importance of the TONSL in 61 embryonic development and postnatal growth. recessive hypomorphic and loss-of- function (LoF) mutations in the TONSL (MIM: 604546) gene of individuals with SPONASTRIME dysplasia. In further studies, using the dermal fibroblasts from individuals, in vitro cell-based assays, in silico struture simulation, and in vivo knock-in mouse model, we demonstrated the pathogenicity of TONSL variants, suggesting that defects in replication-associated DNA damage repair and the resultant inefficient cell proliferation due to TONSL mutations might be the underlying pathogenic mechanism for dysplasia. cloning. investigate the splicing changes caused by the splicing site and deep intronic mutations in P11, reverse transcription-PCR and cloning of the amplicon was performed. The mRNA was harvested from the circulating leukocytes of proband and parents using QIAamp RNA Blood Mini Kit (Qiagen, The cDNA were transcribed using Transcriptor First Strand cDNA Synthesis Kit and then PCR amplification was carried out using the primers and TONSL11R 5’- which encompass both paternal and maternal mutations. After PCR based cloning using an All in One™ PCR Cloning Kit (Biofact, Korea), 30 colonies were picked for PCR and sequencing analyses using the same primers.