Serotonin as a biomarker of toxin-induced Parkinsonism.

BACKGROUND: Loss of dopaminergic neurons underlies the motor symptoms of Parkinson's disease (PD). However stereotypical PD symptoms only manifest after approximately 80% of dopamine neurons have died making dopamine-related motor phenotypes unreliable markers of the earlier stages of the disease. There are other non-motor symptoms, such as depression, that may present decades before motor symptoms. METHODS: Because serotonin is implicated in depression, here we use niche, fast electrochemistry paired with mathematical modelling and machine learning to, for the first time, robustly evaluate serotonin neurochemistry in vivo in real time in a toxicological model of Parkinsonism, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). RESULTS: Mice treated with acute MPTP had lower concentrations of in vivo, evoked and ambient serotonin in the hippocampus, consistent with the clinical comorbidity of depression with PD. These mice did not chemically respond to SSRI, as strongly as control animals did, following the clinical literature showing that antidepressant success during PD is highly variable. Following L-DOPA administration, using a novel machine learning analysis tool, we observed a dynamic shift from evoked serotonin release in the hippocampus to dopamine release. We hypothesize that this finding shows, in real time, that serotonergic neurons uptake L-DOPA and produce dopamine at the expense of serotonin, supporting the significant clinical correlation between L-DOPA and depression. Finally, we found that this post L-DOPA dopamine release was less regulated, staying in the synapse for longer. This finding is perhaps due to lack of autoreceptor control and may provide a ground from which to study L-DOPA induced dyskinesia. CONCLUSIONS: These results validate key prior hypotheses about the roles of serotonin during PD and open an avenue to study to potentially improve therapeutics for levodopa-induced dyskinesia and depression.

Biofiltration of n-butyl acetate with three packing material mixtures, with and without biochar.

Two cost-effective packing materials were used for n-butyl acetate removal in lab-scale biofilters, namely waste spruce root wood chips and biochar obtained as a byproduct from a wood gasifier. Three biofilters packed with spruce root wood chips: without biochar (SRWC), a similar one with 10% of biochar (SRWC-B) and that with 10% of biochar impregnated with a nitrogen fertilizer (SRWC-IB) showed similar yet differing maximum elimination capacities of 206 ± 27, 275 ± 21 and 294 ± 20 g m-3 h-1, respectively, enabling high pollutant removal efficiency (>95% at moderate loads) and stable performance. The original biochar adsorption capacity was high (208 ± 6 mgtoluene g-1), but near 70% of it was lost after a 300-day biofilter operation. By contrast, the exposed impregnated biochar drastically increased its adsorption capacity in 300 days (149 ± 7 vs. 17 ± 5 mgtoluene g-1). Colony forming unit (CFU) and microscopic analyses revealed significant packing material colonization by microorganisms and grazing fauna in all three biofilters with an acceptable pressure drop, up to 1020 Pa m-1, at the end of biofilter operation. Despite a higher price (14 vs. 123 €m-3), the application of the best performing SRWC-IB packing can reduce the total investment costs by 9% due to biofilter volume reduction.

Interrater Reliability for Classifying Craniofacial Microsomia Severity: A Call for Objective Evaluation.

OBJECTIVE: The severity of craniofacial microsomia (CFM) is generally classified using the Orbit, Mandible, Ear, Soft tissue, Nerve (OMENS) classification score. The global assessment of the Phenotypic Assessment Tool for Craniofacial Microsomia (PAT-CFM), is a pictorial modification of the OMENS classification. The aim of this study was to assess the interrater reliability of the PAT-CFM global assessment score. DESIGN: In this prospective cohort study, three clinicians completed the global assessment form of the PAT-CFM. The mandible was classified based on orthopantomogram- and/or computed tomography images. PARTICIPANTS: Consecutive patients with CFM or microtia.Interrater agreement was calculated using the weighted Krippendorff alpha (α), with 95% confidence intervals (CI). RESULTS: In total, 53 patients were included (106 hemifaces). The reliabilities of the main classification components ranged from high for the mandible (α = 0.904 [95% CI 0.860-0.948]) and ear (α = 0.958 [95% CI 0.934-0.983]) subscales, to tentative for the orbital summary score (α = 0.682 [0.542-0.821]), and nerve summary score (α = 0.782 [0.666-0.900]) subscales. CONCLUSIONS: The reliability of the ear and radiographic mandible scales of the PAT-CFM global classification were high, while the orbit, facial nerve and soft tissue subscales may have limited reliability. Research focusing on radiographic severity scores for hypoplasia of the orbits and soft tissues, as well as objective measures for overall facial hypoplasia using non-ionizing forms of imaging for early classification, are warranted.

aEEG vs cEEG's sensivity for seizure detection in the setting of neonatal intensive care units: A systematic review and meta-analysis.

AIM: Amplitude-integrated electroencephalography (aEEG)'s accuracy compared to conventional electroencephalography (cEEG) has not been fully established. The aim of our study was to conduct a systematic review on the sensitivity of the aEEG for neonatal seizure detection. METHODS: Studies from PubMed and Google Scholar databases comparing recordings of cEEG and aEEG in newborns were included according to the PRISMA method. A quality assessment using the QUADAS-2 tool was provided. A random-effect model was used to account for different sources of variations among studies. Publication biases were represented by a funnel plot, and funnel plot symmetry was assessed. RESULTS: Fourteen studies were reported; sensitivity of each diagnostic tool used (single-channel aEEG, two-channel aEEG, two-channel aEEG plus raw trace EEG) was compared to that of the gold-standard cEEG and to those of the other methods used. Overall sensitivity of the aEEG ranged from 31.25% to 90%. CONCLUSION: Our study provides evidence that sensitivity of aEEG varies significantly and that seizure detection rate is lower than that of cEEG. The two-channel aEEG with raw trace EEG shows a high sensitivity and might represent a valid alternative to the cEEG in the setting of neonatal intensive care units (NICUs).

COVID 19 vaccine distribution solution to the last mile challenge: Experimental and simulation studies of ultra-low temperature refrigeration system.

Most COVID-19 vaccines require ambient temperature control for transportation and storage. Both Pfizer and Moderna vaccines are based on mRNA and lipid nanoparticles requiring low temperature storage. The Pfizer vaccine requires ultra-low temperature storage (between -80 °C and -60 °C), while the Moderna vaccine requires -30 °C storage. Pfizer has designed a reusable package for transportation and storage that can keep the vaccine at the target temperature for 10 days. However, the last stage of distribution is quite challenging, especially for rural or suburban areas, where local towns, pharmacy chains and hospitals may not have the infrastructure required to store the vaccine. Also, the need for a large amount of ultra-low temperature refrigeration equipment in a short time period creates tremendous pressure on the equipment suppliers. In addition, there is limited data available to address ancillary challenges of the distribution framework for both transportation and storage stages. As such, there is a need for a quick, effective, secure, and safe solution to mitigate the challenges faced by vaccine distribution logistics. The study proposes an effective, secure, and safe ultra-low temperature refrigeration solution to resolve the vaccine distribution last mile challenge. The approach is to utilize commercially available products, such as refrigeration container units, and retrofit them to meet the vaccine storage temperature requirement. Both experimental and simulation studies are conducted to evaluate the technical merits of this solution with the ability to control temperature at -30 °C or -70 °C as part of the last mile supply chain for vaccine candidates.

In silico MS/MS spectra for identifying unknowns: a critical examination using CFM-ID algorithms and ENTACT mixture samples.

High-resolution mass spectrometry (HRMS) enables rapid chemical annotation via accurate mass measurements and matching of experimentally derived spectra with reference spectra. Reference libraries are generated from chemical standards and are therefore limited in size relative to known chemical space. To address this limitation, in silico spectra (i.e., MS/MS or MS2 spectra), predicted via Competitive Fragmentation Modeling-ID (CFM-ID) algorithms, were generated for compounds within the U.S. Environmental Protection Agency's (EPA) Distributed Structure-Searchable Toxicity (DSSTox) database (totaling, at the time of analysis, ~ 765,000 substances). Experimental spectra from EPA's Non-Targeted Analysis Collaborative Trial (ENTACT) mixtures (n = 10) were then used to evaluate the performance of the in silico spectra. Overall, MS2 spectra were acquired for 377 unique compounds from the ENTACT mixtures. Approximately 53% of these compounds were correctly identified using a commercial reference library, whereas up to 50% were correctly identified as the top hit using the in silico library. Together, the reference and in silico libraries were able to correctly identify 73% of the 377 ENTACT substances. When using the in silico spectra for candidate filtering, an examination of binary classifiers showed a true positive rate (TPR) of 0.90 associated with false positive rates (FPRs) of 0.10 to 0.85, depending on the sample and method of candidate filtering. Taken together, these findings show the abilities of in silico spectra to correctly identify true positives in complex samples (at rates comparable to those observed with reference spectra), and efficiently filter large numbers of potential false positives from further consideration. Graphical abstract.

CFM9, a Mitochondrial CRM Protein, Is Crucial for Mitochondrial Intron Splicing, Mitochondria Function and Arabidopsis Growth and Stress Responses.

Although the importance of chloroplast RNA splicing and ribosome maturation (CRM) domain-containing proteins has been established for chloroplast RNA metabolism and plant development, the functional role of CRM proteins in mitochondria remains largely unknown. Here, we investigated the role of a mitochondria-targeted CRM protein (At3g27550), named CFM9, in Arabidopsis thaliana. Confocal analysis revealed that CFM9 is localized in mitochondria. The cfm9 mutant exhibited delayed seed germination, retarded growth and shorter height compared with the wild type under normal conditions. The growth-defect phenotypes were more manifested upon high salinity, dehydration or ABA application. Complementation lines expressing CFM9 in the mutant background fully recovered the wild-type phenotypes. Notably, the mutant had abnormal mitochondria, increased hydrogen peroxide and reduced respiration activity, implying that CFM9 is indispensable for normal mitochondrial function. More important, the splicing of many intron-containing genes in mitochondria was defective in the mutant, suggesting that CFM9 plays a crucial role in the splicing of mitochondrial introns. Collectively, our results provide clear evidence emphasizing that CFM9 is an essential factor in the splicing of mitochondrial introns, which is crucial for mitochondrial biogenesis and function and the growth and development of Arabidopsis.

Correlative microscopy methods that maximize specimen fidelity and data completeness, and improve molecular localization capabilities.

Correlative microscopy techniques interrogate biological systems more thoroughly than is possible using a single modality. This is particularly true if disparate data types can be acquired from the same specimen. Recently, there has been significant progress towards combining the structural information obtained from soft X-ray tomography (SXT) with molecular localization data. Here we will compare methods for determining the position of molecules in a cell viewed by SXT, including direct visualization using electron dense labels, and by indirect methods, such as fluorescence microscopy and high numerical aperture cryo-light microscopy. We will also discuss available options for preserving the in vivo structure and organization of the specimen during multi-modal data collection, and how some simple specimen mounting concepts can ensure maximal data completeness in correlative imaging experiments.

Novel analytical techniques for HIV-1 infection of CD4 + T cells on fractional order in mathematical biology.

In this research, we present an analytical analysis of HIV-1 infection of CD4 + T cells with a conformable derivative model (CDM) in biology. An improved Υ'/Υ-expansion method is used to investigate this model analytically to construct a new exact traveling wave solution, namely, exponential function, trigonometric function, and the hyperbolic function, which can be further studied for more (FNEE) fractional nonlinear evolution equations in biology. Also, we provide some graphs in 2D plots that demonstrate how accurate the results will be produced using analytical approaches.

A Family Systems Perspective of Elder Family Financial Exploitation: Examining Family Context Profiles.

OBJECTIVE: Informed by Family Systems Theory, this study examines the interwoven nature of intergenerational family structures and roles in which elder family financial exploitation (EFFE) takes place. METHOD: Data from a mixed-methods study were utilized to examine patterns of family structures and roles within and across 23 family systems. Participants were non-perpetrator/non-victim concerned family members (CFMs) who had experienced EFFE. RESULTS: Four family context profiles were identified, organized by the number and roles of victim(s) and perpetrator(s) in each family system, including: Single victim, Single perpetrator; Single victim, Multiple perpetrators; Two victims, Single perpetrator; and Two victims, Multiple perpetrators. It is possible for multiple perpetrators to be involved, with up to five perpetrators across three generations in our sample. DISCUSSION: Findings offer insight into the range of intergenerational family structures and roles affected by EFFE. Practitioners are encouraged to understand and address family contexts to improve EFFE intervention outcomes.

Can commonly-used fan-driven air cleaning technologies improve indoor air quality? A literature review.

Air cleaning techniques have been applied worldwide with the goal of improving indoor air quality. The effectiveness of applying these techniques varies widely, and pollutant removal efficiency is usually determined in controlled laboratory environments which may not be realized in practice. Some air cleaners are largely ineffective, and some produce harmful by-products. To summarize what is known regarding the effectiveness of fan-driven air cleaning technologies, a state-of-the-art review of the scientific literature was undertaken by a multidisciplinary panel of experts from Europe, North America, and Asia with expertise in air cleaning, aerosol science, medicine, chemistry and ventilation. The effects on health were not examined. Over 26,000 articles were identified in major literature databases; 400 were selected as being relevant based on their titles and abstracts by the first two authors, who further reduced the number of articles to 160 based on the full texts. These articles were reviewed by the panel using predefined inclusion criteria during their first meeting. Additions were also made by the panel. Of these, 133 articles were finally selected for detailed review. Each article was assessed independently by two members of the panel and then judged by the entire panel during a consensus meeting. During this process 59 articles were deemed conclusive and their results were used for final reporting at their second meeting. The conclusions are that: (1) None of the reviewed technologies was able to effectively remove all indoor pollutants and many were found to generate undesirable by-products during operation. (2) Particle filtration and sorption of gaseous pollutants were among the most effective air cleaning technologies, but there is insufficient information regarding long-term performance and proper maintenance. (3) The existing data make it difficult to extract information such as Clean Air Delivery Rate (CADR), which represents a common benchmark for comparing the performance of different air cleaning technologies. (4) To compare and select suitable indoor air cleaning devices, a labeling system accounting for characteristics such as CADR, energy consumption, volume, harmful by-products, and life span is necessary. For that purpose, a standard test room and condition should be built and studied. (5) Although there is evidence that some air cleaning technologies improve indoor air quality, further research is needed before any of them can be confidently recommended for use in indoor environments.

Ginkwanghols A and B, osteogenic coumaric acid-aliphatic alcohol hybrids from the leaves of Ginkgo biloba.

Ginkgo biloba (Ginkgoaceae), commonly known as "ginkgo", is called a living fossil, and it has been cultivated early in human history for various uses in traditional medicine and as a source of food. As part of ongoing research to explore the chemical diversity and biologically active compounds from natural resources, two new coumaric acid-aliphatic alcohol hybrids, ginkwanghols A (1) and B (2) were isolated from the leaves of G. biloba. The coumaric acid-aliphatic alcohol hybrids of natural products have rarely been reported. The structures of the new compounds were determined by extensive NMR spectroscopic analysis, HRESI-MS, and quantum chemical ECD calculations, and by comparing the experimental HRESI-MS/MS spectrum of chemically transformed compound 1a with the predicted HRESI-MS/MS spectra proposed from CFM-ID 3.0, a software tool for MS/MS spectral prediction and MS-based compound identification. Ginkwanghols A (1) and B (2) increased alkaline phosphatase (ALP) production in C3H10T1/2, a mouse mesenchymal stem cell line, in a dose-dependent manner. In addition, ginkwanghols A and B mediated the promotion of osteogenic differentiation as indicated by the induction of the mRNA expression of the osteogenic markers ALP and osteopontin (OPN).

Telmisartan Facilitates the Anticancer Effects of CARP-1 Functional Mimetic and Sorafenib in Rociletinib Resistant Non-small Cell Lung Cancer.

BACKGROUND/AIM: Tyrosine kinase inhibitors (TKIs) are used for the treatment of both wild type and mutant non-small cell lung cancer (NSCLC); however, acquired resistance is a major clinical challenge. Herein, we aimed to investigate the effects of telmisartan (Tel), CFM 4.16 and sorafenib combination in rociletinib resistant NSCLC tumors. MATERIALS AND METHODS: 3D spheroid cultures and western blotting were used for evaluating cytotoxic effects and protein expression. An in vivo rociletinib resistant H1975 xenograft model of NSCLC was developed by subcutaneous injection of rociletinib resistant H1975 cells into nude mice. RESULTS: Tel, CFM 4.16 and sorafenib combination displayed superior anti-cancer effects in 3D spheroid cultures and a rociletinib resistant H1975 xenograft model of NSCLC by decreasing the protein expression of oncogenic and cancer stem cell markers (Nanog, Sox2 and Oct4). CONCLUSION: Tel facilitates effective penetration of CFM 4.16 and sorafenib in rociletinib resistant H1975 models of NSCLC.

Measuring child functioning: Assessing correlation and agreement between caregiver and child responses at the Iganga-Mayuge health and demographic surveillance site in Uganda.

INTRODUCTION: Empirical data is scare on assessment of concordance between caregiver-child responses on child functioning. OBJECTIVE: To assess correlation and agreement between children (11-17 years old) and their caregivers' responses to the UNICEF/Washington Group Child Functioning Module (CFM) at the Iganga-Mayuge Health and Demographic Surveillance Site (IM-HDSS) in Uganda. METHODS: CFM with 24-questions corresponding to 13 domains of functioning was administered to children between 11 and 17 years of age and their caregivers. Descriptive analyses of the child/caregiver responses were conducted. Correlation and agreement between caregiver and child responses were assessed. RESULTS: Of the 217 caregiver/child pairs eligible for this study, 181 pairs agreed to participate (83.4%). The mean age of children was 13.9 ± 1.9 years, and 56.4% were males. Cronbach's alpha was 0.892 and 0.886 for the caregiver and child versions of CFM respectively, showing good internal consistency in both. There was a significant overall agreement between mean score of caregiver (5.36 ± 5.63 out of 39) and child (5.45 ± 5.34) pairs. Spearman's rank correlation between the pairs was 0.806 (strong positive correlation). Bland-Altman plots for CFM scores showed greater agreement between caregiver and child at lower scores. Percentage agreement between the pairs for overall disability was greater for mild (83.53%) and moderate (79.37%) categories as compared to the severe (66.67%) category. There was substantial agreement (kappa 0.623) for overall disability between the pairs. CONCLUSION: This study indicates that there is significant correlation and agreement between self-reported caregiver-child pair responses, opening the way for considering children as CFM respondents, when possible.

The Space-Time Coupled Fractional Cattaneo-Friedrich Maxwell Model with Caputo Derivatives.

In the current article, we have thoroughly investigated the collective impact of mixed convection with thermal radiation and chemical reaction on MHD flow of viscous and electrically conducting fluid (Cattaneo-Friedrich Maxwell-CFM model) over a permeable surface embedded in a porous medium. Here we have utilized the Caputo time-fractional derivatives and mechanical laws (generalized shear stress constitutive equation and generalized Fourier's and Fick's laws) are being used to fractionalize the presented model. The effects of radiative heat flux, Ohmic dissipation, and internal absorption are presented through generalized Fourier's law while Fick's law or mass transfer equation offers the effects of first order chemically reactive species. The finite element method and finite difference method are being utilized to numerically solve the nonlinear coupled differential equations. It is established, through compression of numerical and analytical solutions, that the presented model is convergent. Further, error analysis of the subject model is also carried out. Moreover, for better illustration of results, we have also offered a graphical and tabular presentation of impacts of the parameters of interest on velocity, temperature, concentration profile, local skin friction coefficient, and heat and mass transfer. It is evident from the obtained results that velocity near and away from the surface increases with the enhancement of fractional derivative parameter whereas an opposite trend is observed in the case of temperature. Furthermore, it is noticed that temperature shows a decreasing behavior for the value Λ Î¸ < 2 and Λ Ï• < 2 , on the other hand entirely opposite trend is witnessed for Λ Î¸ ≥ 3 and Λ Ï• ≥ 3 . From an engineering perspective, we have acquired comprehensive outcomes such that the heat transfer offers an increasing trend in the case of T R and thermal fractional parameter ß 1 . Additionally, the chemical reaction parameter and Sc significantly contribute towards the mass transfer rate. Since, in literature, one cannot refer to such results with non-integer Caputo fractional derivatives thus the results obtained through the current assessment hold significance for future research avenues. Moreover, the numerical inferences of the subject study may contribute to an advanced thermal processing method in the food industry to swiftly increase the temperature for cooking or sterilization drives.

Characterization and Fabrication of the CFM-JTE for 4H-SiC Power Device with High-Efficiency Protection and Increased JTE Dose Tolerance Window.

A 13.5 kV 4H-SiC PiN rectifier with a considerable active area of 0.1 cm2 is fabricated in this paper. Charge-field-modulated junction termination extension (CFM-JTE) has been proposed for satisfying the requirement of ultra-high reverse voltage, which enlarges the JTE dose tolerance window, making it approximately 2.8 times that of the conventional two-zone JTE. Besides, the CFM-JTE can be implemented through the conventional two-zone JTE process. The measured forward current is up to 100 A @ VF = 5.2 V in the absence of carrier lifetime enhancement technology. The CFM-JTE structure accomplishes 96% of the theoretical breakdown voltage of the parallel plane junction with a relatively small terminal area of 400 µm, which contributes to achieving the Baliga's figure of merit of 58.8 GW/cm2.

Clinical investigation of the utility of a pair of coagulation-fibrinolysis markers for definite diagnosis of sepsis-induced disseminated intravascular coagulation: A single-center, diagnostic, prospective, observational study.

BACKGROUND: Disseminated intravascular coagulation (DIC) often occurs with sepsis. A scoring system has been used for the diagnosis of DIC, but the system included at least 4 parameters. The purpose of this study was to propose a simple set of DIC criteria with coagulation-fibrinolysis markers (CFMs). METHODS: Patients with ≥2 signs of systemic inflammatory response syndrome and a quick Sequential Organ Failure Assessment score ≥ 2 points were investigated. All blood samples were collected on Days 0, 1, 3, and 7. Cutoff values of CFMs were calculated by receiver operating curve analysis. Positive predictive values (PPVs) and negative predictive values (NPVs) for the Japanese Association for Acute Medicine (JAAM) DIC criteria were evaluated by pairing the markers. Differences were analyzed by the Mann-Whitney U test, Kruskal-Wallis test, and the log-rank test. RESULTS: A total of 107 patients were enrolled. The cutoff values of soluble fibrin (SF), protein C (PC), and plasminogen activator inhibitor (PAI)-1 were 48 µg/mL, 42%, and 71 ng/mL according to the International Society of Thrombosis and Hemostasis DIC criteria. The PPV of the severe SFxPC group was 100% for the JAAM DIC criteria, excluding Day 0. CONCLUSION: Cutoff values of SF over 48 µg/mL and PC <42% could almost definitely identify JAAM DIC.

The adsorption of chlorofluoromethane on pristine, and Al- and Ga-doped boron nitride nanosheets: a DFT, NBO, and QTAIM study.

In the present investigation, the feasibility of detecting the chlorofluoromethane (CFM) gas molecule onto the outer surface of pristine single layer boron nitride nanosheet (BNNS), as well as its aluminum (Al)- and gallium (Ga)-doped structures, was carefully evaluated. For achieving this goal, a density functional theory level of study using the Perdew, Burke, and Ernzerhof exchange-correlation (PBEPBE) functional together with a 6-311G(d) basis set has been used. Subsequently, the B3LYP, CAM-B3LYP, wB97XD, and M062X functionals with a 6-311G(d) basis set were also employed to consider the single-point energies. Natural bond orbital (NBO) and quantum theory of atoms in molecules (QTAIM) were implemented by using the B3LYP-D3/6-311G(d) method, and the results were compatible with the electronic properties. In this regard, the total density of states (TDOSs), the Wiberg bond index (WBI), natural charge, natural electron configuration, donor-acceptor natural bond orbital interactions, and the second-order perturbation energies are performed to explore the nature of the intermolecular interactions. All of the energy calculations and population analyses denote that by adsorbing of the gas molecule onto the surface of the considered nanostructures, the intermolecular interactions are of the type of strong chemical adsorption. Among the doped nanosheets, Ga-doped nanosheet has very high adsorption energy compared with other elements (i.e., Ga-doped > Al-doped > pristine). Generally, it was revealed that the sensitivity of the adsorption will be increased when the gas molecule interacts with decorated nanosheets and decrease the HOMO-LUMO band gap; therefore, the change of electronic properties can be used to design suitable nanosensors to detect CFM gas. Graphical abstract.

The bRPS6-Family Protein RFC3 Prevents Interference by the Splicing Factor CFM3b during Plastid rRNA Biogenesis in Arabidopsis thaliana.

Plastid ribosome biogenesis is important for plant growth and development. REGULATOR OF FATTY ACID COMPOSITION3 (RFC3) is a member of the bacterial ribosomal protein S6 family and is important for lateral root development. rfc3-2 dramatically reduces the plastid rRNA level and produces lateral roots that lack stem cells. In this study, we isolated a suppressor of rfc three2 (sprt2) mutant that enabled recovery of most rfc3 mutant phenotypes, including abnormal primary and lateral root development and reduced plastid rRNA level. Northern blotting showed that immature and mature plastid rRNA levels were reduced, with the exception of an early 23S rRNA intermediate, in rfc3-2 mutants. These changes were recovered in rfc3-2 sprt2-1 mutants, but a second defect in the processing of 16S rRNA appeared in this line. The results suggest that rfc3 mutants may be defective in at least two steps of plastid rRNA processing, one of which is specifically affected by the sprt2-1 mutation. sprt2-1 mutants had a mutation in CRM FAMILY MEMBER 3b (CFM3b), which encodes a plastid-localized splicing factor. A bimolecular fluorescence complementation (BiFC) assay suggested that RFC3 and SPRT2/CFM3b interact with each other in plastids. These results suggest that RFC3 suppresses the nonspecific action of SPRT2/CFM3b and improves the accuracy of plastid rRNA processing.

Novel strategies to target chemoresistant triple-negative breast cancer.

Previous studies from our group and others have shown that current drug treatment(s) strategies eliminate bulk of tumor cells (non-CSCs) but it had a minimal effect on cancer stem cells (CSCs) leading to resistance and tumor recurrence. We studied the effects of CFM-4.16 (CARP-1 functional mimetic) and/or cisplatin on four Triple-negative breast cancer (TNBC) MDA-MB-468, MDA-MB-231, CRL-2335 and BR-1126, two cisplatin resistant CisR/MDA-231 and CisR/MDA-468 and cancer stem cells (CSCs) from resistant cell lines. TNBC cells treated with CFM-4.16 plus cisplatin inhibited the expression of FZD8, LRP6 and c-Myc and significantly enhanced cell death in all the cell lines by ~70%-80% compared with the control(s). When Cisplatin resistant CisR/MDA-231 and CisR/MDA-468 were treated with CFM-4.16 plus cisplatin, they also showed a reduction in FZD8 and LRP6 and increased apoptosis compared to control group. Similarly, CFM-4.16 plus cisplatin treatment reduced mammospheres formation abilities of CSCs by 80-90% compared to control group, increased PARP cleavage and apoptosis. Data shows CFM-4.16 plus cisplatin treatment significantly increased apoptosis/cell death in parental, cisplatin resistant and CSCs. Taken together the data suggests that FZD8-mediated Wnt-signaling plays a major role in mediating CSCs growth and resistance to chemotherapy and its inhibition enhances the chemotherapeutic response in TNBC.