This study aimed to estimate the prevalence of, and factors associated with, accessing maternal healthcare services (MHC) by Indigenous women in the Chittagong Hill Tracts (CHT), Bangladesh.

Sporadic literature reports describe isolates of pathogenic bacteria that harbor an antibiotic resistance determinant but remain susceptible to the corresponding antibiotic as a consequence of a genetic defect. Such strains represent a source from which antibiotic resistance may reemerge to cause treatment failure in patients. Here, we report a systematic investigation into the prevalence and nature of this phenomenon, which we term silencing of antibiotic resistance by mutation (SARM). Instances of SARM were detected among 1,470 Staphylococcus aureus isolates through side-by-side comparison of antibiotic resistance genotype (as determined by whole-genome sequencing) versus phenotype (as assessed through susceptibility testing). Of the isolates analyzed, 152 (10.3%) harbored a silenced resistance gene, including 46 (3.1%) that exhibited SARM to currently deployed antistaphylococcal drugs. SARM resulted from diverse mutational events but most commonly through frameshift mutation of resistance determinants as a result of point deletion in poly(A) tracts. The majority (∼90%) of SARM strains reverted to antibiotic resistance at frequencies of ≥10-9; thus, while appearing antibiotic sensitive in the clinical microbiology laboratory, most S. aureus isolates exhibiting SARM will revert to antibiotic resistance at frequencies achievable in patients. In view of its prevalence in a major pathogen, SARM represents a significant potential threat to the therapeutic efficacy of antibiotics.IMPORTANCE Antibiotic resistance hinders the treatment of bacterial infection. To guide effective therapy, clinical microbiology laboratories routinely perform susceptibility testing to determine the antibiotic sensitivity of an infecting pathogen. This approach relies on the assumption that it can reliably distinguish bacteria capable of expressing antibiotic resistance in patients, an idea challenged by the present study. We report that the important human pathogen Staphylococcus aureus frequently carries antibiotic resistance genes that have become inactivated ("silenced") by mutation, leading strains to appear antibiotic sensitive. However, resistance can rapidly reemerge in most such cases, at frequencies readily achievable in infected patients. Silent antibiotic resistance is therefore prevalent, transient, and evades routine detection, rendering it a significant potential threat to antibacterial chemotherapy.

Tuberculosis, one of the world's most severe infectious diseases, is caused by Mycobacterium tuberculosis A major weapon of this pathogen is a unique cell wall that protects the pathogen from eradication by the immune system. Mycobacteria have specialized secretion systems, e.g., type VII secretion or ESX systems, to transport substrates across this cell wall. The largest group of proteins that are secreted by these ESX systems are the PE proteins. Previously, it was shown that the N-terminal PE domain of about 100 amino acids is required for secretion. Here, we describe the identification of an aspartic protease, designated PecA, that removes (part of) this PE domain at the cell surface. Nearly all of the observed PE_PGRS proteins are processed by PecA. Interestingly, the protease itself is also a secreted PE protein and subject to self-cleavage. Furthermore, a defect in surface processing has no effect on the activity of the PE lipase protein LipY but does seem to affect the functioning of other virulence factors, as a pecA mutant strain of Mycobacterium marinum shows moderate attenuation in zebrafish larvae. In conclusion, our results reveal the presence of a functional aspartic acid protease in M. marinum that cleaves LipY, itself as well as other members of the PE_PGRS family. Finally, mutants lacking PecA show growth attenuation in vivo, suggesting that PecA plays a role during infection.IMPORTANCE Aspartic proteases are common in eukaryotes and retroviruses but are relatively rare among bacteria (N. D. Rawlings and A. Bateman, BMC Genomics 10:437, 2009, https://doi.org/10.1186/1471-2164-10-437). In contrast to eukaryotic aspartic proteases, bacterial aspartic proteases are generally located in the cytoplasm. We have identified a surface-associated mycobacterial aspartic protease, PecA, which cleaves itself and many other type VII secretion substrates of the PE_PGRS family. PecA is present in most pathogenic mycobacterial species, including M. tuberculosis In addition, pathogenicity of M. marinum is reduced in the ΔpecA mutant, indicating that PecA contributes to virulence.

Bacteria express numerous efflux transporters that confer resistance to diverse toxicants present in their environment. Due to a high level of functional redundancy of these transporters, it is difficult to identify those that are of most importance in conferring resistance to specific compounds. The resistance-nodulation-division (RND) protein family is one such example of redundant transporters that are widespread among Gram-negative bacteria. Within this family, the MexAB-OprM protein complex is highly expressed and conserved among Pseudomonas species. We exposed barcoded transposon mutant libraries in isogenic wild-type and ΔmexB backgrounds in P. syringae B728a to diverse toxic compounds in vitro to identify mutants with increased susceptibility to these compounds. Mutants with mutations in genes encoding both known and novel redundant transporters but with partially overlapping substrate specificities were observed in a ΔmexB background. Psyr_0228, an uncharacterized member of the major facilitator superfamily of transporters, preferentially contributes to tolerance of acridine orange and acriflavine. Another transporter located in the inner membrane, Psyr_0541, contributes to tolerance of acriflavine and berberine. The presence of multiple redundant, genomically encoded efflux transporters appears to enable bacterial strains to tolerate a diversity of environmental toxins. This genome-wide screen performed in a hypersusceptible mutant strain revealed numerous transporters that would otherwise be dispensable under these conditions. Bacterial strains such as P. syringae that likely encounter diverse toxins in their environment, such as in association with many different plant species, probably benefit from possessing multiple redundant transporters that enable versatility with respect to toleration of novel toxicants.IMPORTANCE Bacteria use protein pumps to remove toxic compounds from the cell interior, enabling survival in diverse environments. These protein pumps can be highly redundant, making their targeted examination difficult. In this study, we exposed mutant populations of Pseudomonas syringae to diverse toxicants to identify pumps that contributed to survival in those conditions. In parallel, we examined pump redundancy by testing mutants of a population lacking the primary efflux transporter responsible for toxin tolerance. We identified partial substrate overlap for redundant transporters, as well as several pumps that appeared more substrate specific. For bacteria that are found in diverse environments, having multiple, partially redundant efflux pumps likely allows flexibility in habitat colonization.

Homologous sequence alignments contain important information about the constraints that shape protein family evolution. Correlated changes between different residues, for instance, can be highly predictive of physical contacts within three-dimensional structures. Detecting such co-evolutionary signals via direct coupling analysis is particularly challenging given the shared phylogenetic history and uneven sampling of different lineages from which protein sequences are derived. Current best practices for mitigating such effects include sequence-identity-based weighting of input sequences and post-hoc re-scaling of evolutionary coupling scores. However, numerous weighting schemes have been previously developed for other applications, and it is unknown whether any of these schemes may better account for phylogenetic artifacts in evolutionary coupling analyses. Here, we show across a dataset of 150 diverse protein families that the current best practices out-perform several alternative sequence- and tree-based weighting methods. Nevertheless, we find that sequence weighting in general provides only a minor benefit relative to post-hoc transformations that re-scale the derived evolutionary couplings. While our findings do not rule out the possibility that an as-yet-untested weighting method may show improved results, the similar predictive accuracies that we observe across distinct weighting methods suggests that there may be little room for further improvement on top of existing strategies.

A new species, Callistethus hamus Lu & Zorn, sp. nov., is described from China, Laos, and Vietnam. Additionally, we used synchrotron (Shanghai Synchrotron Radiation Facility) to scan the aedeagus. The virtual 3D model of the aedeagus is reconstructed and provided.

Roots of Solanum lycopersicum L. were collected in growing season of year 2015, on the island of Santiago in Cape Verde. Morphological, morphometric and molecular (18S rDNA and 28S rDNA) studies revealed the presence of Pratylenchus brachyurus and P. delattrei in root systems and root zones of tomato plants. To our knowledge, this is the first record of the occurrence of these nematode species in Cape Verde.

The trematode Postlepidapedon opisthobifurcatum (Zdzitowiecki, 1990) is a common intestinal parasite of the gadiform fishes of the Southern Ocean. In this work, we supplement the description of the species with the anatomy of the terminal part of the reproductive system and with molecular data. The male terminal genitalia are characterised by the presence of the external seminal vesicle and cirrus-sac. The external seminal vesicle is surrounded by aciniform groups of outer prostatic cells. Groups of outer prostatic cells and proximal parts of their ducts are associated with a thin-walled membrane that is connected to the proximal edge of the сirrus-sac. The cirrus-sac is claviform, with a long proximal part accommodating the tubular, thin-walled internal seminal vesicle and ducts of outer prostatic cells. The female terminal genitalia are represented by a thick-walled metraterm, which is surrounded by aciniform groups of glandular cells. Phylogenetic analysis based on 28S rDNA partial sequences data placed P. opisthobifurcatum into the monophyletic group Lepidapedidae, including the species Myzoxenus insolens (Crowcroft, 1945), Intusatrium robustum Durio et Manter, 1968, and Postlepidapedon uberis Bray, Cribb et Barker, 1997. However, we were unable to detect direct phylogenetic connections between P. opisthobifurcatum and P. uberis.

Dactylogyrid monogeneans of Labeo horie Heckel, 1847 and L. niloticus (Linnaeus, 1758) (Cyprinidae) were surveyed at two sites on the River Nile in Sudan. The present study reports the presence of ten species of Dactylogyrus Diesing, 1850 and three species of Dogielius Bychowsky, 1936 based on a morphometric evaluation of the sclerotised structures. The species found include: Dactylogyrus nathaliae Guégan, Lambert & Euzet, 1988; D. rastellus Guégan, Lambert & Euzet, 1988; D. retroversus Guégan, Lambert & Euzet, 1988; D. senegalensis Paperna, 1969, D. yassensis Musilová, Řehulková & Gelnar, 2009 and five other undescribed species of Dactylogyrus. The genus Dogielius was represented by Dogielius flosculus Guégan, Lambert & Euzet, 1989; the newly identified D. sennarensis n. sp., and one undescribed Dogielius species. While D. sennarensis n. sp. resembles D. intorquens, it differs from this species and other congeners by having a longer ventral bar and anchor points and nosclerotised vagina. In addition, this study redescribes D. flosculus based on the morphology of specimens collected from L. horie. All specimens studied had a single large vagina, in contrast to the original description which reported a vagina composed of two unconnected parts. All dactylogyrid species in this study represent new host and geographical records.

To study the effect of fertilization on soil nematode communities in a paddy-upland rotation system, an ongoing thirty-three years long-term fertilizer experiment is conducted which includes seven treatments; an unfertilized treatment (control), nitrogen (N), phosphorus (P), potassium (K) fertilizer treatments (N, NP, NPK) and organic manure (M) combined with chemical fertilizer treatments (MN, MNP, and MNPK). The soil nematode community structure and crop yields were determined in 2012 and 2013. Overall total nematode abundance was increased by an incremental nutrient input both in the rice and wheat fields. Total number of nematode was 1.25 - 2.37 times greater in the rice field and was 1.08 - 2.97 times greater in wheat field in the fertilization treatments than in the unfertilized treatment. Soil free-living nematode abundances was significantly (P < 0.001) increased in organic manure combined with chemical fertilizer treatments in rice field. Fungi-feeders and plant-feeding nematodes abundances were not significantly different among treatments in rice and wheat fields. Omnivorous and predatory nematodes were the most dominant groups in the present study. Omnivores, predators and Prodorylaimus abundances were significantly (P < 0.05) higher in organic manure combined with NPK fertilizer treatments than in chemical fertilizer alone and unfertilized treatments both in rice and wheat fields. Stepwise regressions revealed that soil free-living nematodes were significant predictors of rice grain yields (R 2 = 0.56, P < 0.001) and omnivorous and predatory nematodes were significant predictors of the wheat grain yield (R 2 = 0.89, P < 0.001). Therefore, long-term application of organic manure combined with chemical fertilizer could increase nematode abundances and crop yields. Organic manure combined with chemical fertilizer application was recommended in agricultural ecosystem.

We morphologically and molecularly characterized segmented filamentous bacteria (SFB) associated with Rhigonema sp. nematodes in millipede hindguts. Seventy-three Riukiaria sp. millipedes were collected from a broad-leaf forest in Japan, and nematodes were excised from the millipede's hindguts. The occurrence rate of SFB associated with nematodes was 24 % (10/41) for males, 47 % (14/30) for females, and 100 % (2/2) for juveniles. Genomic DNA was extracted from four SFB-rich nematode heads, and we obtained 40 bacterial clones via analysis of nearly full-length 16S rDNA gene sequences. At the phylum level, Firmicutes, Proteobacteria, and Verrucomicrobia accounted for 55 %, 40 %, and 5 % of SFB, respectively. In Firmicutes, Clostridiaceae (28 %) and Lachnospiraceae (15 %) were the dominant groups. Our sequences were divided into seven and three subclades between Firmicutes and Proteobacteria in the phylogenetic tree. In the Firmicutes clade, eight sequences were classified as Lachnospiraceae with a bootstrap value >83 %. A phylogenetic tree involving known uncultured Lachnospiraceae sequences characterized the phylogenetic position of SFB associated with nematodes. Our results suggest that the association of SFB with nematode bodies was probably incidental and that SFB are not always present in millipede hindguts. Our bacterial groups corresponded to those of arthropod hindgut, and SFB associated with nematodes were inferred to belong to Lachnospiraceae. Because the Lachnospiraceae sequences obtained in this study showed specific lineages that differed from all the known deposited sequence data, these groups may be unique to Riukiaria sp.

Lifestyle medicine (LM) is part of official educational goals in Swedish medical schools. We studied questions concerning 5 noncommunicable diseases: diabetes, hypertension, coronary heart disease (CHD), chronic obstructive pulmonary disease (COPD), and stroke from 124 written examinations conducted between 2012 and 2015. LM knowledge yielded between 2% and 10%, whereas pharmacology-related knowledge yielded between 24% and 50%, of total points. The multiples at which pharmacology-related knowledge was valued higher than LM knowledge were 2.4 for COPD (P < .056), 4.3 for diabetes (P < .0001), 4.8 for hypertension (P < .0001), 5.2 for CHD (P < .0001), and 31.5 for stroke (P < .0001). Our results indicate that lifestyle-related knowledge, though covered by official teaching goals, is currently underrated in Swedish medical education.

To assess the periodontal status among the leather factory workers in Dewas and to provide a baseline data for the oral health care and promotion programs.

Achilles tendon pathologies occur frequently and have a significant socioeconomic impact. Currently, there is no evidence on the best treatment for these pathologies. Cell therapy has been studied in several animal models, and encouraging results have been observed with respect to tissue regeneration. This study is aimed at evaluating the functional and histological effects of bone marrow stem cell or platelet-rich plasma implantation compared to eccentric training in the treatment of Achilles tendinopathy in rats.

The current study investigates the inhibitory effects of Pulsatilla pentacyclic triterpenoid saponins extract (PPTS) on epithelial-mesenchymal transition (EMT) triggered by the transforming growth factor-β1 (TGF-β1) in human colorectal cancer SW480 cell line, further illustrates the possible mechanism of PPTS inhibition of growth and invasion from the perspective of EMT, and provides new theoretical support for the treatment of tumor by Chinese medicine. The SW480 cells were treated in groups: blank control, TGF-β1 (10 ng/mL), and varying concentrations of PPTS cotreated with TGF-β1-induced (10 ng/mL) groups. CCK8 was used to detect cell viability; transwell was applied to detect invasion ability, cell migration ability was also determined, ELISA and RT-qPCR were utilized for the determination of CYP3A, CYP2C9, CYP2C19, N-cadherin, and MMP-9 expression. Flow cytometry detection was applied to detect cell cycle and apoptosis. The results obtained have shown that PPTS can significantly inhibit the invasion and migration of tumors in SW480 cells and can also block the S phase in the cell cycle but may produce cytotoxicity in higher doses. The present research work provides substantial evidence that PPTS has a significant inhibitory effect on TGF-β1-induced EMT in SW480 cells and it also promotes apoptosis.

Nowadays, an increasing number of patients are seriously affected by lung cancer. Si Jun Zi Tang (SJZ), a four-herb Chinese medicine formula first described approximately one thousand years ago, is often prescribed for cancer patients as a complementary therapy. But the research on the effective materials for treating cancer using SJZ was rarely reported. To solve this problem, we evaluate the inhibitory effect of 10 samples of SJZ from different origins on PC9 cells. Ultraperformance liquid chromatography (UPLC) and hierarchical cluster analysis (HCA) along with canonical correlation analysis (CCA) and bioactivity validation were used to investigate the underlying correlation between the chemical ingredients and the antiproliferative effect of SJZ on PC9 cells. The evaluation indicated that 10 batches of SJZ could inhibit proliferation of PC9 cells and there was a notable difference in pharmacological activity between the different SJZ samples. The results of CCA and multivariate statistical analysis indicated that ginsenoside Ro and ginsenoside Rg1 might be active constituents of the antiproliferative effect as determined by spectrum-effect relationships. The results showed that bioassay and spectrum-effect relationships are suitable to associate sample quality with the active ingredient associated with clinical efficacy. And our finding would provide foundation and further understanding of the quality evaluation of traditional Chinese medicine decoction.

To investigate how a back propagation neural network based on genetic algorithm (GA-BPNN) optimizes the low-intensity pulsed ultrasound (LIPUS) stimulation parameters to improve the bone marrow mesenchymal stem cells (BMSCs) viability further. The LIPUS parameters were set at various frequencies (0.6, 0.8, 1.0, and 1.2 MHz), voltages (5, 6, 7, and 8 V), and stimulation durations (3, 6, and 9 minutes). As only some discrete points can be set up in the experiments, the optimal LIPUS stimulation parameter may not be in the value of these settings. The GA-BPNN algorithm is used to optimize parameters of LIPUS to increase the BMSCs viability further. The BMSCs viability of the LIPUS-treated group was improved up to 19.57% (P < 0.01). With the optimization via the GA-BPNN algorithm, the viability of BMSCs was further improved by about 5.36% (P < 0.01) under the optimized condition of 6.92 V, 1.02 MHz, and 7.3 min. LIPUS is able to improve the BMSCs viability, which can be improved further by LIPUS with parameter optimization via GA-BPNN algorithm.

Based on previous observations that the nutraceutical CELLFOOD™ (CF), the 'physiological modulator' that aimed to make oxygen available 'on demand', inhibits the growth of cancer cells, this study was designed to investigate the role of CF in the regulation of hypoxia-inducible factor 1 alpha (HIF1α) and its correlated proteins, phosphoglycerate kinase 1 and vascular endothelial growth factor. Our idea was that CF, acting on HIF1α, in combination with current anticancer therapies could improve their effectiveness.

We previously demonstrated increased expression of programmed cell death 5 (PDCD5) in asthmatic patients and ovalbumin-induced allergic asthma. International guidelines (GINA 2019) have included the use of tiotropium bromide for chronic treatment of the most severe and frequently exacerbated asthma in patients ≥6 years old, who do not have good response to inhaled corticosteroids.

Ectopic expression of miRNAs promotes tumor development and progression. miRNA (miR)-320a is downregulated in many cancers, including gastric cancer (GC). However, the mechanism underlying its downregulation and the role of miR-320a in GC are unknown.