Development and Validation of a Clinical Nomogram for Predicting Complications From Pediatric Multiple Magnet Ingestion: A Large Retrospective Study.

INTRODUCTION: This study aimed to develop and validate a reliable nomogram based on clinical factors to predict complications associated with pediatric multiple magnet ingestion, addressing the urgency and controversy surrounding its management. METHODS: Patients aged 0-18 years with multiple magnet ingestion diagnosed at the Shenzhen Children's Hospital between January 2017 and December 2023 were enrolled. Clinical data were analyzed using least absolute shrinkage and selection operator regression and multifactor logistic regression analyses to screen for risk factors. A model was constructed, and a nomogram was plotted. Model performance was evaluated and internally validated using the area under the curve (AUC), Hosmer-Lemeshow test, calibration curve, decision curve analysis, and 1,000 bootstraps. We calculated the optimal cutoff value, sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the prediction model. RESULTS: Of the 146 patients, 57 (39.0%) experienced complications. The nomogram included age, multiple ingestions, vomiting, abdominal pain, and abdominal tenderness. The AUC was 0.941, and the internally validated AUC was 0.930. The optimal cutoff value selected as a predictive value was 0.534, with a sensitivity of 82.5%, specificity of 93.3%, positive predictive value of 88.7%, negative predictive value of 89.3%, and accuracy of 89.0%. The Hosmer-Lemeshow test yielded a P value of 0.750. The calibration plot exhibited high consistency in prediction, and decision curve analysis showed excellent net benefits. DISCUSSION: Our nomogram demonstrates excellent discrimination, calibration, and clinical utility and may thus help clinicians accurately assess the risk of complications from pediatric multiple magnet ingestion.

Mechanical and thermal properties of graphyne-coated carbon nanotubes: a molecular dynamics simulation on one-dimensional all-carbon van der Waals heterostructures.

The mechanical and thermal properties of a hybrid nanotube consisting of a coaxial carbon nanotube (CNT) inside a graphyne nanotube (GNT), i.e., CNT@GNT, are investigated in this paper by using molecular dynamics simulations. The results show that the mechanical properties of CNT@GNT under uniaxial tension depend on the nanotube chirality of its components. Specifically, the Young's modulus of the CNT@GNT structure with an inner zigzag CNT is larger than that of its counterpart with an armchair CNT, while CNT@GNT with an armchair CNT and a zigzag GNT is found to possess the largest tensile strength and fracture strain. In addition, a unique fracture behavior of the successive rupture of its two components is observed in CNT@GNT. The thermal conductivity of CNT@GNT is found to be almost independent of the nanotube chirality of its components but increases as the length and diameter of the CNT@GNT increase. Moreover, strain engineering is shown as an effective avenue to modulate the thermal conductivity of CNT@GNT, which can be enhanced by tension but reduced by compression. The analysis of the phonon spectrum and spectral energy density demonstrates that this strain effect originates from changes of the phonon group velocity and phonon scattering in the strained CNT@GNT.

Mechanical force regulates root resorption in rats through RANKL and OPG.

BACKGROUND: External root resorption is one of common complications of orthodontic treatment, while internal root resorption is rarely observed, and the difference between pulp and periodontal tissues during orthodontic treatment is still unknown. The purpose of this study was to evaluate the effects of orthodontic forces on histological and cellular changes of the dental pulp and periodontal tissues. METHODS: Orthodontic tooth movement model was established in Forty-eight adult male Wistar rats. The distance of orthodontic tooth movement was quantitatively analyzed. The histological changes of pulp and periodontal tissues were performed by hematoxylin-eosin staining, tartrate-resistant acid phosphate staining was used to analyze the changes of osteoclast number, immunohistochemistry analysis and reverse transcription polymerase chain reaction were used to examine the receptor of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG) expression. The width of tertiary dentine was quantitatively analyzed. Tartrate-resistant acid phosphate staining and the erosion area of osteo assay surface plate was used to evaluate osteoclast activity. RESULTS: The orthodontic tooth movement distance increased in a force dependent manner, and reached the peak value when orthodontic force is 60 g. Heavy orthodontic force increased the RANKL expression of periodontal ligament srem cells (PDLSCs) which further activated osteoclasts and resulted in external root resorption, while the RANKL expression of dental pulp stem cells (DPSCs) was relatively low to activate osteoclasts and result in internal root resorption, and the dental pulp tend to form tertiary dentine under orthodontic force stimulation. CONCLUSIONS: Heavy orthodontic forces activated osteoclasts and triggered external root resorption by upregulating RANKL expression in rat periodontal tissues, while there was no significant change of RANKL expression in dental pulp tissue under heavy orthodontic forces, which prevented osteoclast activation and internal root resorption.

An rfuABCD-Like Operon and Its Relationship to Riboflavin Utilization and Mammalian Infectivity by Borrelia burgdorferi.

Riboflavin is an essential micronutrient, but its transport and utilization have remained largely understudied among pathogenic spirochetes. Here, we show that Borrelia burgdorferi, the zoonotic spirochete that causes Lyme disease, is able to import riboflavin via products of its rfuABCD-like operon as well as synthesize flavin mononucleotide and flavin adenine dinucleotide despite lacking canonical genes for their synthesis. Additionally, a mutant deficient in the rfuABCD-like operon is resistant to the antimicrobial effect of roseoflavin, a natural riboflavin analog, and is attenuated in a murine model of Lyme borreliosis. Our combined results indicate not only that are riboflavin and the maintenance of flavin pools essential for B. burgdorferi growth but also that flavin utilization and its downstream products (e.g., flavoproteins) may play a more prominent role in B. burgdorferi pathogenesis than previously appreciated.

Knockdown of Long Noncoding RNAs of Maternally Expressed 3 Alleviates Hyperoxia-Induced Lung Injury via Inhibiting Thioredoxin-Interacting Protein-Mediated Pyroptosis by Binding to miR-18a.

Long-term hyperoxia exposure may cause lung damage with characteristic inflammation. Long noncoding RNA of maternally expressed 3 (MEG3) is up-regulated in lung tissues exposed to hyperoxia; however, the underlying mechanism is unclear. Hyperoxia-induced cells and mouse models were used to study these mechanisms. Molecular assays were used to detect cell viability, cytotoxicity, and expression of miR-18a, MEG3, and inflammatory cytokines. The interaction among MEG3, miR-18a, and thioredoxin-interacting protein (TXNIP) was verified; and pyroptosis-related proteins were analyzed. The in vivo model was established by exposing MEG3 knockdown mice to hyperoxia. Hematoxylin and eosin staining was used to assess pathologic alterations of lung tissues. Hyperoxia suppressed cell viability, induced cell damage, and exacerbated the secretion of IL-1ß and IL-18. Hyperoxia inhibited miR-18a, with increased expression of MEG3, TXNIP, and nonobese diabetic-like receptor family pyrin domain containing 3 (NLRP3). MEG3 aggravated TXNIP expression by binding to miR-18a. Knockdown of MEG3 rescued hyperoxia-induced pyroptosis by up-regulating miR-18a. Furthermore, knockdown of MEG3 inhibited NLRP3 inflammasome activity and caspase-1 signaling by miR-18a. In vivo knockdown of MEG3 and overexpression of miR-18a relieved hyperoxia-induced lung injury via restraining NLRP3 inflammasome-mediated pyroptosis, whereas miR-18a inhibition reversed these effects. In conclusion, knockdown of MEG3 inhibits pyroptosis to alleviate hyperoxia lung injury by suppressing NLRP3 inflammasome and caspase-1 signaling via regulating miR-18a-TXNIP axis.

Higher Levels of Triglyceride, Fatty Acid Translocase, and Toll-Like Receptor 4 and Lower Level of HDL-C in Pregnant Women with GDM and Their Close Correlation with Neonatal Weight.

OBJECTIVES: In this study, we aimed to compare the levels of maternal blood lipids, placental and venous blood lipid transporters, and inflammatory factor receptors in pregnant women with and without gestational diabetes mellitus (GDM). We also aimed to figure out the relationship between these values and neonatal weight. METHODS: Fifty pregnant women with GDM under blood glucose control belong to the case group, and 50 pregnant women with normal glucose tolerance in concurrent delivery belong to the control group. Fasting venous blood of these pregnant women was taken 2 weeks before delivery, and umbilical cord blood was collected after delivery. The levels of triglyceride (TG), serum total cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol (HDL-C) in maternal blood and umbilical cord blood were tested in the laboratory department of our hospital. The level of toll-like receptor 4 (TLR4) in serum of umbilical veins was detected by the double-antibody sandwich ELISA. Western blot and RT-PCR were used to detect the protein and mRNA expressions of TLR4, LPL, and FAT/CD36 in the placenta. RESULTS: The level of TG in maternal blood in the case group was remarkably higher than that in the control group, which was opposite to the level of HDL-C. In the umbilical cord blood of women with GDM, the expression of TLR4 increased and was closely correlated with neonatal weight. In the placenta of women with GDM, the expressions of FAT/CD36 and TLR4 increased, and both of them were closely correlated with neonatal weight. Besides, TLR4 in umbilical cord blood increased and was closely correlated with neonatal weight. Although the expression of LPL in the placenta decreased, it had no obvious correlation with neonatal weight. CONCLUSIONS: TG in maternal blood, TLR4 in the placenta and umbilical cord blood, and FAT/CD36 in the placenta were positively correlated with neonatal weight. However, HDL-C in maternal blood was negatively correlated with neonatal weight. Although the expression of LPL in the placenta reduced due to GDM, it had no correlation with neonatal weight.

Integrated eco-strategies towards sustainable carbon and nitrogen cycling in agriculture.

To meet the ever-growing human demands for food, fuel, and fiber, agricultural activities have dramatically altered the global carbon (C) and nitrogen (N) cycles. These biogeochemical cycles along with water, phosphorus, and sulfur cycles are fundamental features of life on Earth. Human alteration of the global N cycle has had both positive and negative outcomes. To efficiently feed a growing population, crop-livestock production systems have been developed, however, these systems also contribute significantly to environmental pollution and global climate change. Management of agricultural waste (AW) and the application of N fertilizers are central to the issues of greenhouse gas (GHG) emissions and nutrient runoff that contributes to the eutrophication of water bodies. If managed properly, AW can provide nutrients for plants and contribute to the conservation of soil health. In order to achieve the long-term conservation of agricultural production systems, it is important to promote the proper recycling of AW in agroecosystems and to minimize the reliance on chemical N fertilizers. Composting is one of the sustainable and effective approaches for recycling AW in agriculture. However, the conventional composting process is dilatory and produces compost with low N content compared to chemical N fertilizers. For this reason, comprehensive research is required to improve the composting process and the N content of the soil organic amendments. This work aims to explore the beneficial effects of the integrated application of biochar and specific C and N cycling microorganisms to the composting process and the quality of the composted products. In pursuit of replacing chemical N fertilizers with bio/organic fertilizers, we further discussed the power of the combined application of compost, biochar, and N-fixing bacteria in agricultural production systems. The knowledge of smart integration of AW and microorganisms in agriculture could solve the main agricultural and environmental problems associated with human-induced flows of C and N. Building upon the knowledge disseminated in review to further extensive research will pave the way for better management of agricultural production systems and sustainable C and N cycling in agriculture.

Effects of interlayer interactions on the nanoindentation response of freely suspended multilayer gallium telluride.

Freestanding indentation is a widely used method to characterise the elastic properties of two-dimensional (2D) materials. However, many controversies and confusion remain in this field due to the lack of appropriate theoretical models in describing the indentation responses of 2D materials. Taking the multilayer gallium telluride (GaTe) as an example, in this paper we conduct a series of experiments and simulations to achieve a comprehensive understanding of its freestanding indentation behaviours. Specifically, the freestanding indentation experiments show that the elastic properties of the present multilayer GaTe with a relatively large thickness can only be extracted from the bending stage in the indentation process rather than the stretching stage widely utilised in the previous studies on thin 2D materials, since the stretching stage of thick 2D materials is inevitably accompanied with severe plastic deformations. In combination with existing continuum mechanical models and finite element simulations, an extremely small Young's modulus of multilayer GaTe is obtained from the nanoindentation experiments, which is two orders of magnitude smaller than the value obtained from first principles calculations. Our molecular dynamics (MD) simulations reveal that this small Young's modulus can be attributed to the significant elastic softening in the multilayer GaTe with increasing thickness and decreasing length. It is further revealed in MD simulations that this size-induced elastic softening originates from the synergistic effects of interlayer compression and interlayer shearing in the multilayer GaTe, both of which, however, are ignored in the existing indentation models. To consider these effects of interlayer interactions in the theoretical modelling of the freestanding indentation of multilayer GaTe, we propose here novel multiple-beam and multiple-plate models, which are found to agree well with MD results without any additional parameters fitting and thus can be treated as more precise theoretical models in characterising the freestanding indentation behaviours of 2D materials.

Altered Expressions of AQP3 and ADP Are Closely Related with the Risk of Preeclampsia Occurrence.

To investigate the expression of aquaporin 3 (AQP3) in placenta and fetal membranes and the level of adiponectin (ADP) in the umbilical cord blood of severe preeclampsia and to analyze the relationship between the 2 proteins and severe preeclampsia, 60 pregnant women with severe preeclampsia were recruited as the case group and another group of 60 normal pregnant women in the same gestation period were selected as the control. After parturition, the transcriptional levels of AQP3 mRNA in placenta and fetal membranes were evaluated with RT-PCR. The expressions of AQP3 protein in the placenta and fetal membranes were determined by immunohistochemistry and Western blot. Meanwhile, the expression of ADP in umbilical artery blood was detected by ELISA. The content of triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and LDL-C/high-density lipoprotein cholesterol in the case group was significantly higher than that in the control group. Multivariate logistic regression analysis showed that the level of TG was related to the onset of severe preeclampsia (OR 2.589). The relative expression of AQP3 mRNA and expression of AQP3 protein in placenta tissue of the case group were significantly lower than those of the control group, whereas in fetal membranes, they were significantly higher. The results of ELISA showed that the level of ADP in umbilical cord blood of neonates in the case group increased remarkably compared with that of the control group.

Biochar-bacteria-plant partnerships: Eco-solutions for tackling heavy metal pollution.

Over the past 30 years, the ever-rising demands of the modern and growing population have led to the rapid development of agricultural and industrial sectors worldwide. However, this expansion has exposed the environment to various pollutants including heavy metal (HM)s. Almost all HMs are serious toxicants and can pose serious health risks to living organisms in addition to their bioaccumulative and non-biodegradable nature. Different techniques have been developed to restore the ecological functions of the HM-contaminated soil (HMCS). However, the major downfalls of the commonly used remediation technologies are the generation of secondary wastes, high operating costs, and high energy consumption. Phytoremediation is a prominent approach that is more innocuous than the existing remediation approaches. Some microbes-plant interactions enhance the bioremediation process, with heavy metal resistant-plant growth promoting bacteria (HMRPGPB) being widely used to assist phytoremediation of HMs. However, the most common of all major microbial assisted-phytoremediation disturbances is that the HM-contaminated soil is generally deficient in nutrients and cannot sustain the rapid growth of the applied HMRPGPB. In this case, biochar has recently been approved as a potential carrier of microbial agents. The biochar-HMRPGPB-plant association could provide a promising green approach to remediate HM-polluted sites. Therefore, this review addresses the mechanisms through which biochar and HMRPGPB can enhance phytoremediation. This knowledge of biochar-HMRPGPB-plant interactions is significant with respect to sustainable management of the HM-polluted environment in terms of both ecology and economy, and it offers the possibility of further development of new green technologies.

Increasing soil moisture faciliates the outcomes of exogenous sulfate rather than element sulfur in reducing cadmium accumulation in rice (Oryza sativa L.).

Cadmium (Cd) contamination in paddy soils and the related pollution risk of rice grain have received increasing attention. Agronomic measures, such as the application of sulfur and changes in water regimes, were reported to mitigate the accumulation of Cd in rice. However, there is limited information on the combined effects of sulfur application and water regimes. Therefore, a pot experiment was conducted to investigate the effects of two sulfur forms, three water regimes and multiple sulfur application rates on Cd accumulation in rice. The sulfur was applied as SO42- (SVI, replacing the traditional fertilizers by SO42--containing fertilizers), and element S (S0) was applied at 0, 10, 20, 30 and 40 mg S kg-1 soil. The water regimes were continuous flooding (F), flooding-moist alternation (FM), and moist irrigation (M), for a total of 30 treatments. The results indicated that application of SVI exceeding 30 mg S kg-1 significantly reduced the Cd concentrations in brown rice by 31.1-56.3%, and the Cd concentrations decreased with increasing amount of irrigation water. Similar reductions in Cd concentrations in rice shoots and rice straw collected at tillering and maturity stages were observed after application of SVI. However, the effect of S0 application on Cd accumulation in grain was not significant under different water regimes. Furthermore, this study found that application of both SVI and S0 inhibited the transfer of Cd from rice roots to shoots in most cases. These findings indicate that replacing traditional fertilizers with SO42--containing fertilizers, especially combined with increased irrigation, could be a potential approach to mitigate Cd accumulation in rice growing in Cd-contaminated acidic paddy soils.

The E3 ubiquitin ligase RNF114 and TAB1 degradation are required for maternal-to-zygotic transition.

The functional role of the ubiquitin-proteasome pathway during maternal-to-zygotic transition (MZT) remains to be elucidated. Here we show that the E3 ubiquitin ligase, Rnf114, is highly expressed in mouse oocytes and that knockdown of Rnf114 inhibits development beyond the two-cell stage. To study the underlying mechanism, we identify its candidate substrates using a 9,000-protein microarray and validate them using an in vitro ubiquitination system. We show that five substrates could be degraded by RNF114-mediated ubiquitination, including TAB1. Furthermore, the degradation of TAB1 in mouse early embryos is required for MZT, most likely because it activates the NF-κB pathway. Taken together, our study uncovers that RNF114-mediated ubiquitination and degradation of TAB1 activate the NF-κB pathway during MZT, and thus directly link maternal clearance to early embryo development.

The putative Walker A and Walker B motifs of Rrp2 are required for the growth of Borrelia burgdorferi.

Rrp2 encodes a putative bacterial enhancer binding protein (bEBP) in Borrelia burgdorferi. Point mutation (G239C) of Rrp2 abolishes the transcriptional activation of σ54 -dependent rpoS. In contrast to canonical bEBPs that are dispensable for bacterial growth, Rrp2 is essential for borrelial growth in BSK medium. It has been believed that Rrp2's ATPase activity is not required for cell growth, but experimental evidence supporting this notion has been lacking. In particular, it has remained unclear whether the residue G239 is involved in Rrp2's presumptive ATPase activity. To address these information gaps, we examined the roles of Rrp2's potential strategic signatures including the G239 residue and the putative Walker A and Walker B motifs. Herein it was showed that Rrp2 has ATP binding and hydrolysis activities engendered by the Walker A and B motifs respectively. However, these activities were not significantly impaired by a G239C mutation. Further mutagenesis analyses indicated that Rrp2's Walker A and B motifs are required for borrelial growth; mutations of key residues in these two motifs were lethal to B. burgdorferi. The combined data suggest that the Walker A and Walker B motifs of Rrp2 are involved in the control of another unknown RpoS-independent gene product(s) associated with borrelial replication.

Piezoelectric effects on the resonance frequencies of boron nitride nanosheets.

By using molecular dynamics (MD) simulations, we find in this work that due to the piezoelectric characteristic of boron nitride (BN) nanosheets their resonance frequencies can be efficiently tuned by applying an external electric field. This finding suggests that BN nanosheet can be treated as a good building block for designing novel piezoelectrically tunable two-dimensional nanoresonators. As BN nanosheets possess an inversely stacked structure, the applied electric field has different effects on the resonance frequency of BN nanosheets with odd and even layers. The influence of piezoelectric effect on the vibration behaviours observed in MD simulations is found to significantly deviate from the prediction of the conventional Euler-Bernoulli beam model (EBM), since the EBM cannot account for the weak van der Waals interaction between neighbouring layers in BN nanosheets. To take into account the interlayer interaction in the mathematical modelling of the piezoelectric effect on the vibration of BN nanosheets, we propose here a novel multiple beam model (MBM), which can account for both interlayer stretching and shearing deformations. The MBM result is found to be in a good agreement with the MD result without any additional parameters fitting, which indicates that the present MBM can be treated as a more precise theoretical model in the future study of the vibration properties of BN nanosheets.

Spontaneous curling of freestanding Janus monolayer transition-metal dichalcogenides.

In this paper, using molecular dynamics simulations we report spontaneous curling behaviors of freestanding Janus monolayer S-Mo-Se (MoSeS) structures. Density functional theory calculations are performed to obtain the phonon dispersion and phonon spectra of the Janus monolayer MoSeS for analyzing its structural stability. The results show that the Janus monolayer MoSeS is structurally stable. Due to the lattice mismatch between MoS and MoSe domains, the Janus monolayer MoSeS at the freestanding state always spontaneously rolls up in a constant temperature and pressure system. The direction of curling is preferred along the armchair orientation. Specifically, as for the Janus monolayer MoSeS whose size is larger than ∼30 nm, it can spontaneously roll up into a nanotube structure. The underlying physical mechanisms of these phenomena are well uncovered by using classical Timoshenko plate theory and the minimum energy principle.

[A case of congenital bile acid synthesis disorder type 2 and literature review].

OBJECTIVE: To summarize the clinical features, biochemical change and genetic mutations of a neonate with congenital bile acid synthesis disorder type 2. METHODS: Clinical features, blood biochemical index, gene analysis and treatment of the patient were reviewed. RESULTS: The patient presented with the symptoms of jaundice 3 days after birth but without skin itching. Pale stool was noted. Subsequently, he presented with hepatomegaly, blood coagulation disorders, left cochlear nerve damage, liver cirrhosis and remarkable growth retardation. Serum biochemistries showed that bilirubin and transaminase were elevated, while γ -GT and total bile acid was normal. Abdominal ultrasonography indicated decline of gallbladder contraction. Cholangiography showed normal extra- and intrahepatic bile ducts and patent biliary tract. Liver biopsy showed intrahepatic cholestasis. Gene testing has identified a homozygous mutation in AKR1D1 gene. CONCLUSION: Congenital bile acid synthesis disorder should be suspected when a neonate has presented with jaundice, elevated bilirubin and transaminase, normal or reduced TBA and γ -GT. Genetic testing and urine mass spectrometry analysis can diagnose congenital bile acid synthesis disorder. Early therapy is crucial to patients with congenital bile acid synthesis disorder.

Nepetaefolins A-J, Cytotoxic Chinane and Abietane Diterpenoids from Caryopteris nepetaefolia.

Nepetaefolins A-J (1-10) and seven known compounds were isolated from the whole plant of Caryopteris nepetaefolia. The absolute configurations of 1-3 were determined from single-crystal X-ray diffraction and spectroscopic data. Compounds 6 and 7, with IC50 values of 6.3-9.0 µM, showed higher cytotoxicity than paclitaxel in one non-small-cell lung cancer, patient-derived xenograft (PDX) model when tested using PDX models and the adenosine triphosphate-tumor chemosensitivity assay (ATP-TCA).

Identification of four cornua by ultra-performance liquid chromatography with time-of-flight mass spectrometry coupled with principal component analysis.

An ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry method coupled with principal component analysis was developed and applied to the identification of Cornu Antelopis, Cornu Bubali, Cornu Naemorhedi, and Cornu Bovis. The data obtained from the trypsin-digested samples were subjected to principal component analysis to classify these four cornua. Additionally, marker peptides of the cornua were determined by orthogonal partial least-squares discriminant analysis, and fragmentation tandem mass spectra of these marker peptides were evaluated. The results from this study indicate that the proposed method is reliable, and it has been successfully applied to the identification of variants of cornua commonly used in traditional Chinese medicine.

Evidence that BosR (BB0647) Is a Positive Autoregulator in Borrelia burgdorferi.

Borrelia burgdorferi survives in nature through a complex tick-mammalian life cycle. During its transit between ticks and mammalian hosts, B. burgdorferi must dramatically alter its outer surface profile in order to interact with and adapt to these two diverse niches. It has been established that the regulator BosR (BB0647) in B. burgdorferi plays important roles in modulating borrelial host adaptation. However, to date, how bosR expression itself is controlled in B. burgdorferi remains largely unknown. Previously, it has been shown that DNA sequences upstream of BosR harbor multiple sites for the binding of recombinant BosR, suggesting that BosR may influence its own expression in B. burgdorferi However, direct experimental evidence supporting this putative autoregulation of BosR has been lacking. Here, we investigated the expression of bosR throughout the tick-mammal life cycle of B. burgdorferi via quantitative reverse transcription (RT)-PCR analyses. Our data indicated that bosR is expressed not only during mouse infection, but also during the tick acquisition, intermolt, and transmission phases. Further investigation revealed that bosR expression in B. burgdorferi is influenced by environmental stimuli, such as temperature shift and pH change. By employing luciferase reporter assays, we also identified two promoters potentially driving bosR transcription. Our study offers strong support for the long-postulated function of BosR as an autoregulator in B. burgdorferi.

BadR (BB0693) controls growth phase-dependent induction of rpoS and bosR in Borrelia burgdorferi via recognizing TAAAATAT motifs.

In Borrelia burgdorferi (Bb), the alternative sigma factor RpoS plays a central role during Bb's adaptation to ticks and mammals. Previous studies have demonstrated that RpoS is not expressed during the early stages of spirochetal growth or when Bb resides in ticks during the intermolt phase, but the molecular details of these events remain unknown. In the current study, biomagnetic bead separation of rpoS promoter-binding proteins, coupled with genetic inactivation, was employed to identify BadR (BB0693) as a negative regulator that controls growth phase-dependent induction of rpoS and bosR in Bb. When badR was inactivated, the expression of rpoS and bosR was induced only during the early stages of bacterial growth, but not during the stationary growth phase. Recombinant BadR bound to the promoter DNA of rpoS and the regulatory region upstream of bosR via AT-rich TAAAATAT motifs. Mutations in this motif markedly inhibited or abolished rBadR binding. These results suggest that BadR directly influences the expression of both rpoS and bosR in Bb. This newly recognized role for BadR to fine-tune the activation of the RpoN-RpoS pathway at strategic times in Bb's life cycle potentially represents another layer of gene control over σ(54)-dependent gene regulation.