Molecular characteristics of Staphylococcus aureus strains isolated from subclinical mastitis of water buffaloes in Guangdong Province, China.

Intramammary infections (IMI) in animals reared for milk production can result in large economic losses and distress to the animals. Staphylococcus aureus is an important causative agent of IMI in dairy cows, but its prevalence in water buffaloes has not been determined. Therefore, the current study was conducted to investigate the prevalence of subclinical mastitis in water buffaloes and the antimicrobial susceptibility, virulence genes and biofilm formation abilities of Staphylococcus aureus isolates recovered from water buffaloes in Guangdong, China. Staphylococcus aureus strains were isolated from milk samples of water buffaloes with subclinical mastitis, and twofold microdilution, PCR and crystal violet staining methods were used to determine antimicrobial susceptibility, distributions of virulence and antimicrobial resistance genes and biofilm formation ability, respectively. Our results indicated that 29.44% of water buffaloes were diagnosed with subclinical mastitis, and the most prevalent pathogens were Escherichia coli (96.17%), coagulase-negative staphylococci (CoNS) (67.60%) and S. aureus (28.57%). Most S. aureus isolates showed resistance to bacitracin, doxycycline, penicillin, florfenicol, and tetracycline but were susceptible to ciprofloxacin, ceftizoxime, cefoquinoxime, and ofloxacin. Moreover, 63.72% of S. aureus isolates were positive for tetM, and the prevalence of msrB, blaZ, mecA, fexA, and tetK ranged from 21.24 to 6.19%. All S. aureus isolates harbored clfB and icaA genes, and the virulence genes hla (93.8%), hld (91.15%), clfA (90.27%), fnbA (86.73%), and hlb (83.19%), and tsst, icaD, sec, see, fnbB, and sea showed a varied prevalence ranging from 3.5 to 65.49%. All S. aureus isolates possessed the ability to form biofilms, and 30.09% of isolates showed strong biofilm formation abilities, while 19.47% of isolates were weak biofilm producers. Our results indicated that subclinical mastitis is prevalent in water buffaloes in Guangdong, China, and S. aureus is prevalent in samples from water buffaloes with subclinical mastitis. Most S. aureus isolates were susceptible to cephalosporins and fluoroquinolones; thus, ceftizoxime and cefoquinoxime can be used to treat subclinical mastitis in water buffaloes.

In situ preparation of hydroxyapatite in lamellar liquid crystals for joint lubrication and drug delivery.

Arthritis is a disease that seriously affects the quality of human life, which is partly caused by the reduction of joint lubrication performance. Thus, for the treatment of arthritis, how to improve the lubrication performance of joints is important. The lamellar liquid crystals (LLCs) systems have the potential to be used as joint lubrication due to their double-layer structure and good biocompatibility, however, the LLCs system alone could not provide a satisfactory lubrication effect. Herein, this work synthesized hydroxyapatite (HAP) in situ inside Tween 85/Tween 80/H2O LLCs to construct a biocompatible HAP/Tween 85/Tween 80/H2O LLCs (HAP/LLCs) lubrication system with both sustained drug release properties and anti-wear properties. HAP is the main component of bone with good stability and bioactivity. The LLCs have good lubricating and drug-carrying properties. The impact of HAP on the structure and lubrication properties of LLCs, the mechanism of friction, and the anti-wear reduction of HAP/LLCs were investigated. Moreover, the drug release behavior of the ibuprofen-loaded HAP/LLCs during the friction process was also studied. The results indicated that the addition of HAP could improve the lubricity performance of LLCs. The cumulative drug releasing increased with the friction frequency and was less affected by the load. The related studies provided the theoretical basis for HAP/LLCs for joint lubrication and synergistic therapy.

End-to-End diagnosis of breast biopsy images with transformers.

Diagnostic disagreements among pathologists occur throughout the spectrum of benign to malignant lesions. A computer-aided diagnostic system capable of reducing uncertainties would have important clinical impact. To develop a computer-aided diagnosis method for classifying breast biopsy images into a range of diagnostic categories (benign, atypia, ductal carcinoma in situ, and invasive breast cancer), we introduce a transformer-based hollistic attention network called HATNet. Unlike state-of-the-art histopathological image classification systems that use a two pronged approach, i.e., they first learn local representations using a multi-instance learning framework and then combine these local representations to produce image-level decisions, HATNet streamlines the histopathological image classification pipeline and shows how to learn representations from gigapixel size images end-to-end. HATNet extends the bag-of-words approach and uses self-attention to encode global information, allowing it to learn representations from clinically relevant tissue structures without any explicit supervision. It outperforms the previous best network Y-Net, which uses supervision in the form of tissue-level segmentation masks, by 8%. Importantly, our analysis reveals that HATNet learns representations from clinically relevant structures, and it matches the classification accuracy of 87 U.S. pathologists for this challenging test set.

Lubrication and Dynamically Controlled Drug Release Properties of Tween 85/Tween 80/H2O Lamellar Liquid Crystals.

Lamellar liquid crystals have amazing lubricating and drug-solubilizing properties. Hence, the combination of drug molecules with lamellar liquid crystals is expected to be used in joint lubrication and treatment. In this study, the partial phase diagram of the Tween 85/Tween 80/H2O three-component system was determined. The phase structure of the system changed from a hexagonal liquid crystal to a lamellar liquid crystal with the increase of Tween 85 content. The lamellar liquid crystals showed superior lubricating properties due to their unique lamellar structure. Furthermore, the model of drug release during friction was established for the first time. It was found that the order of the lamellar liquid crystals increased with the increase of the mass ratio of Tween 85/Tween 80, leading to the decrease of the ibuprofen release rate. In addition, the release rate of ibuprofen increased progressively with the increase of the friction frequency, but the load had little effect on it. Therefore, the lamellar liquid crystals consisting of nonionic surfactants with good biocompatibility have potential application prospects for joint lubrication and treatment of arthritis.

Analysis of Regions of Interest and Distractor Regions in Breast Biopsy Images.

This paper studies why pathologists can misdiagnose diagnostically challenging breast biopsy cases, using a data set of 240 whole slide images (WSIs). Three experienced pathologists agreed on a consensus reference ground-truth diagnosis for each slide and also a consensus region of interest (ROI) from which the diagnosis could best be made. A study group of 87 other pathologists then diagnosed test sets (60 slides each) and marked their own regions of interest. Diagnoses and ROIs were categorized such that if on a given slide, their ROI differed from the consensus ROI and their diagnosis was incorrect, that ROI was called a distractor. We used the HATNet transformer-based deep learning classifier to evaluate the visual similarities and differences between the true (consensus) ROIs and the distractors. Results showed high accuracy for both the similarity and difference networks, showcasing the challenging nature of feature classification with breast biopsy images. This study is important in the potential use of its results for teaching pathologists how to diagnose breast biopsy slides.

One-Pot Synthesis of Fe/N-Doped Hollow Carbon Nanospheres with Multienzyme Mimic Activities against Inflammation.

Inflammation, including infectious and noninfectious inflammation, are a growing threat to public health worldwide. For different types of inflammation, more specific and intensified therapy is needed. Nanozymes are able to regulate levels of radical reactive oxygen species (ROS) to suppress inflammation, becoming potential anti-inflammatory agents. Herein, hollow porous carbon spheres codoped with nitrogen and iron (Fe/N-HCNs) are synthesized through a one-pot strategy, which exerted multienzyme mimicking activities, including peroxidase (POD)-, oxidase (OXD)-, catalase (CAT)-, and superoxide dismutase (SOD)-like activities. Moreover, these activities were promoted by the removal of iron oxides produced in the synthesis process. Based on the study of multienzyme activities, we designed two kinds of animal inflammatory models, bacteria-infected wound and inflammatory bowel disease, to evaluate the anti-inflammation ability of Fe/N-HCNs. The results indicated that Fe/N-HCNs could increase ROS levels through performing their POD-like activity in a weak acid environment to catalyze H2O2 against bacteria-infected wound healing, whereas Fe/N-HCNs with the capability of scavenging ROS in a neutral environment could also be unitized to treat noninfectious inflammatory bowel disease. Together, our study provided evidence that the prominent multienzyme activities of Fe/N-HCNs could be used as an anti-inflammatory alternative for both infectious and noninfectious inflammation.

Folding Dynamics of Parallel and Antiparallel G-Triplexes under the Influence of Proximal DNA.

The G-triplex is a kind of DNA structure formed by G-rich sequences. Previous studies have shown that it is an intermediate for the folding of G-quadruplex and has an antiparallel structure. The folding dynamics of this G-triplex structure, however, have not been well studied until now. In addition, whether a parallel G-triplex structure exists, remains unknown. In this study, by using single-molecule fluorescence resonance energy transfer and circular dichroism spectroscopy methods, we have studied the folding dynamics of the G-triplex and revealed at the single-molecule level, for the first time, that G-triplexes have both parallel and antiparallel structures. Moreover, we have investigated the effects of proximal DNA on G-triplex folding. We have found that both single-stranded TTA and double-stranded DNA at either end of a G-triplex sequence can reduce its folding speed. More interestingly, when located at the 5' end of a G-triplex sequence, the proximal DNA will favor the folding of parallel over antiparallel G-triplex structures. As G-triplex is an intermediate for G-quadruplex folding, the present results may also shed new light on the folding properties of G-quadruplex structures, in terms of dynamics, stability, and the effects of proximal DNA.

Effects of monovalent cations on folding kinetics of G-quadruplexes.

G-quadruplexes are special structures existing at the ends of human telomeres, the folding kinetics of which are essential for their functions, such as in the maintenance of genome stability and the protection of chromosome ends. In the present study, we investigated the folding kinetics of G-quadruplex in different monovalent cation environments and determined the detailed kinetic parameters for Na+- and K+-induced G-quadruplex folding, and for its structural transition from the basket-type Na+ form to the hybrid-type K+ form. More interestingly, although Li+ was often used in previous studies of G-quadruplex folding as a control ion supposed to have no effect, we have found that Li+ can actually influence the folding kinetics of both Na+- and K+-induced G-quadruplexes significantly and in different ways, by changing the folding fraction of Na+-induced G-quadruplexes and greatly increasing the folding rates of K+-induced G-quadruplexes. The present study may shed new light on the roles of monovalent cations in G-quadruplex folding and should be useful for further studies of the underlying folding mechanism.

Multimeric BLM is dissociated upon ATP hydrolysis and functions as monomers in resolving DNA structures.

Bloom (BLM) syndrome is an autosomal recessive disorder characterized by an increased risk for many types of cancers. Previous studies have shown that BLM protein forms a hexameric ring structure, but its oligomeric form in DNA unwinding is still not well clarified. In this work, we have used dynamic light scattering and various stopped-flow assays to study the active form and kinetic mechanism of BLM in DNA unwinding. It was found that BLM multimers were dissociated upon ATP hydrolysis. Steady-state and single-turnover kinetic studies revealed that BLM helicase always unwound duplex DNA in the monomeric form under conditions of varying enzyme and ATP concentrations as well as 3'-ssDNA tail lengths, with no sign of oligomerization being discerned. Measurements of ATPase activity further indicated that BLM helicase might still function as monomers in resolving highly structured DNAs such as Holliday junctions and D-loops. These results shed new light on the underlying mechanism of BLM-mediated DNA unwinding and on the molecular and functional basis for the phenotype of heterozygous carriers of BLM syndrome.

[Effect of Cornus officinalis fruit core extract on the cardiac hypertrophy induced by two kidney two clip].

OBJECTIVE: To observe the effect of Cornus officinalis fruit core extract on cardiac hypertrophy induced by two kidney two clip (2K2C) and its mechanism. METHODS: Male Sprague-Dawley rats were randomly divided into 4 groups: sham-operated group, model group and treatment groups (300, 600 mg/kg). Rats were intragastric administered medicine for 4 weeks from the fourth week after surgery. Sham-operated and 2K2C rats were given vehicle for 4 weeks. Blood pressure and hemodynamic parameters were measured. Left ventricular weight to body weight (LVM/BM) ratio was calculated. Paraffin-embedded hearts were cut into 5 microm slices, which were stained with hematoxylin-eosin (HE) and Masson for morphological analysis; Western-blot analysis was performed to investigate the effects of Cornus officinalis fruit core extract on the expression of P47phox, Nox4 in myocardium. RESULTS: Compared with sham-operated group, the blood pressure and LVM/BM ratios were markedly elevated in model groups. Meanwhile cardiomyocyte cross sectional areas was markedly increased and myocardial fibers showed disordered arrangement while these parameters were markedly reversed after treatment with Cornus officinalis fruit core extract for 4 weeks. At 8th weeks after operation, model rats developed obvious LV hypertrophy. Cornus officinalis fruit core extract, more significant in high dose, decreased the blood pressure and LVM/BM ratios and reversed the cardiomyocyte hypertrophy and myocardial fibrosis. Moreover, Cornus officinalis fruit core extract decreased the expression of P47phox and Nox4 which elevated in LV in model rats. CONCLUSION: Cornus officinalis fruit core extract could significantly decrease the blood pressure, reverse cardiac hypertrophy and improve the function of heart which is possibly associated with the down-regulation of P47phox and Nox4.