Search

The central tenet of scientific research is the rigorous application of the scientific method to experimental design, analysis, interpretation, and reporting of results. In order to confer validity to a hypothesis, experimental details must be transparent and results must be reproducible. Failure to achieve this minimum indicates a deficiency in rationale, design, and/or execution, necessitating further experimental refinement or hypothesis reformulation. More importantly, rigorous application of the scientific method advances scientific knowledge by enabling others to identify weaknesses or gaps that can be exploited by new ideas or technology that inevitably extend, improve, or refine a hypothesis. Experimental details, described in manuscript materials and methods, are the principal vehicle used to communicate procedures, techniques, and resources necessary for experimental reproducibility. Recent examination of the biomedical literature has shown that many published articles lack sufficiently detailed methodological information to reproduce experiments. There are few broadly established practice guidelines and quality assurance standards in basic biomedical research. The current paper provides a framework of best practices to address the lack of reporting of detailed materials and methods that is pervasive in histological slide-based assays. Our goal is to establish a structured framework that highlights the key factors necessary for thorough collection of metadata and reporting of slide-based assays. [ABSTRACT FROM AUTHOR]

Investigating the function of delicate mammalian eyes often requires chemical fixation, histological sectioning, immunohistochemistry (IHC) and in situ hybridization (ISH). One of the long-standing challenges in the ocular histology field is the limited success of maintaining intact morphology via cryo- or paraffin procedures. Although our latest protocol significantly improved the morphology of mouse eyeball sections, the window technique is time-consuming and requires extensive practice to avoid damage while making windows. In this study, we present a novel glyoxal fixative that is suitable for a freeze-substitution approach to improve both morphology and molecular target preservation of mouse eyes. The method prevents morphology distortion in all tested eyeballs. Therefore, it suits a variety of research needs from morphological examination to investigation of single-molecule RNA expression, using hematoxylin and eosin (H&E) stain, IHC, and ISH assays on either frozen (cryo) or paraffin-infiltrated tissue sections. In addition, this method can be easily performed in many histology laboratories. [ABSTRACT FROM AUTHOR]

Patients with idiopathic pulmonary fibrosis (IPF) are still suffering from unfavorable survival. BTB and CNC homology1 (Bach1) is a regulator of oxidative stress and participates in the pathogenesis of multiple lung diseases. Thus, this study aimed to determine the effect of Bach1 knockdown on fibrosis and inflammation in pulmonary fibrosis (PF) mice and cell models. Bleomycin induced PF mice were constructed and treated with Bach1 siRNA adenovirus (BLM + Bach1 siRNA group), control siRNA adenovirus (BLM + Control siRNA group) or normal saline (BLM group), then lung tissues were collected for Bach1 expression detection, H&E staining and Masson's trichrome staining. Afterwards, collagen type I alpha 1 chain (COL1A1) and interleukin-6 (IL-6) expressions in serum and bronchoalveolar lavage fluid (BALF) were examined. Subsequently, mouse lung fibroblasts (MLFs) were collected from PF mice and treated with TGF-β1 to construct PF cell model, which was treated with Bach1 siRNA adenovirus (TGF-β1 + Bach1 siRNA group) and MAP kinase (ERK) inhibitor U0126 alone (TGF-β1 + U0126 group) or in combination (TGF-β1 + U0126 + Bach1 siRNA group), then alpha-smooth muscle actin (α-SMA), fibronectin 1 (Fn1), COL1A1, IL-6 expressions and cell viability were detected. Lung tissue Bach1 mRNA and protein expressions were upregulated in PF mice compared to control mice. Bach1 knockdown reduced lung fibrosis (displayed by Masson's trichrome staining) and inflammation (displayed by H&E staining), then downregulated serum and BALF expressions of COL1A1 and IL-6 in PF mice. Subsequently, in PF cell model, Bach1 knockdown blocked ERK pathway, but did not affect Smads, c-Jun N-terminal kinase (JNK) or thymoma viral proto-oncogene 1 (Akt) pathways. Further experiments revealed that Bach1 knockdown repressed cell viability, α-SMA, Fn1, IL-6 and COL1A1 expressions in PF cell model, then ERK inhibition by U0126 enhanced these effects. Bach1 is involved in the PF pathogenesis via modulating ERK signaling pathway. [ABSTRACT FROM AUTHOR]

COVID-19 disease in humans, caused by the novel SARS-CoV-2 virus, was first reported in the city of Wuhan, China in December 2019. This disease has quickly developed into a global pandemic, resulting in over 350,000 deaths worldwide and over 5 million confirmed infections in a matter of 6 months. Although the genome of this novel viral RNA was sequenced quickly and testing kits were manufactured to assist in combatting COVID-19, the diagnosis and treatment will remain relatively unsuccessful until the pathology of this disease is fully understood. Histotechnology plays an important role in understanding the pathology of many diseases, including COVID-19. The first postmortem biopsy of a COVID-19 patient was collected on 27 January 2020, and the pathology finding was published in mid-February 2020. Since then, more studies have been published in scientific literatures as the global outbreak continues. This mini-review summarizes the published articles in which histotechnology aspects were utilized with the intent to understand the pathology of COVID-19. In addition, it is anticipated there will be more molecular and immunohistochemical studies to further understand the mechanism of the disease in the near future. [ABSTRACT FROM AUTHOR]

The BaseScope™ assay is a novel, highly sensitive RNA in situ hybridization (ISH) technique, allowing detection of short RNA sequences as well as discrimination between single-nucleotide alterations. Multiplexing BaseScope™ ISH with immunofluorescence assay has proven challenging because the diffusion of colorimetric dyes such as Fast Red in aqueous solutions degrades spatial resolution. In this study, we explore alkaline phosphatase-based fluorescent signal detection methods and integrate it with BaseScope™ RNA ISH. We found that Fast Blue BB/NAMP can be used in BaseScope™ ISH for signal detection. Additionally, we found that the calcium binding fluorochromes calcein and xylenol orange can be used to localize alkaline phosphatase activity in both immunohistochemistry (IHC) and BaseScope™ ISH assays. When applied to mouse brain and intestine tissue sections, we successfully detected circular RNA molecules and cell proliferation antigen Ki-67 proteins. This technological advance expanded the substrate selection of alkaline phosphatase-based BaseScope™ RNA ISH to allow researchers and clinical professionals accurately assess RNA targets with immunofluorescent multiplexing. [ABSTRACT FROM AUTHOR]

Melanomas represent the malignant transformation of melanocytes, cells found primarily in the skin to protect epithelium and underlying connective tissues from harmful effects of ultraviolet light. Melanomas vary greatly in morphology and may continue to produce melanin markedly, mildly, or not at all. Performing and evaluating hematoxylin and eosin stains as well as immunohistochemical stains on pigmented melanomas has been a long-standing challenge due to the obscuring pigment. Protocols for removing melanin to reveal cellular morphology have been used successfully for years, but coupling these protocols with stains for immunohistochemistry represents an added challenge. In this study, the investigators evaluated results of various melanin bleaching protocols on tissue morphology, completeness of melanin removal, and immunohistochemistry staining quality. It was found that 1% formamide in 3% H2O2 under bright light without heating outperformed other tested protocols. [ABSTRACT FROM AUTHOR]

Objective: To use the finite element model of a Lenke 5 adolescent idiopathic scoliosis (AIS) patient to simulate four corrections (including anterior and posterior correction); to investigate the corrective effect of different surgical protocols; and to analyze the biomechanical stress and strain of the scoliotic spines. Methods: Four surgical strategies were designed and simulated with the model of scoliosis. All the main steps of each strategy, including derotation and compression, were simulated. The stress variation of the spine and the corrective effect were compared among the protocols for different surgical approaches and fusion levels. Results: With the four different surgical protocols, the coronary lumbar deformity was corrected to 22°, 23°, 26° and 26°, respectively, and a physiological sagittal configuration was maintained; however, higher stress was observed with solutions A1 (screw model implanted in the convex side of T12-L3) and A2 (screw model implanted in the convex side of T11-L4), while solution B2 (the posterior approach: T10-L5, fusion to SV) lost too many lumbar movement segments. A similar apical rotational correction was recorded (41.68° and 37.79°) for solutions A2 and B1 (the posterior approach: T10-L4, fusion to LEV), which both instrumented the lower end vertebrae. Conclusions: The presented model could be used successfully to simulate correction procedures, including 90° derotation and compression, for the first time. The Lenke 5 AIS in this particular case was more rigid, and solution B1 was considered the ideal choice for treatment of this patient. [ABSTRACT FROM AUTHOR]

It is established that stress impairs spatial learning and memory via the hypothalamus-pituitary-adrenal axis response. Dopamine D1 receptors were also shown to be responsible for a stress-induced deficit of working memory. However, whether stress affects the subsequent emotional learning and memory is not elucidated yet. Here, we employed the well-established one-trial step-through task to study the effect of an acute psychological stress (induced by tail hanging for 5, 10, or 20 min) on emotional learning and memory, and the possible mechanisms as well. We demonstrated that tail hanging induced an obvious stress response. Either an acute tail-hanging stress or a single dose of intraperitoneally injected dopamine D1 receptor antagonist (SCH23390) significantly decreased the step-through latency in the one-trial step-through task. However, SCH23390 prevented the acute tail-hanging stress-induced decrease in the step-through latency. In addition, the effects of tail-hanging stress and/or SCH23390 on the changes in step-through latency were not through non-memory factors such as nociceptive perception and motor function. Our data indicate that the hyperactivation of dopamine D1 receptors mediated the stress-induced deficit of emotional learning and memory. This study may have clinical significance given that psychological stress is considered to play a role in susceptibility to some mental diseases such as depression and post-traumatic stress disorder. [ABSTRACT FROM AUTHOR]

Journal of Histotechnology (JOH) has something to celebrate with all of you this time of the year. Although hematoxylin and eosin (H&E) and special stained sections are preferred by many pathologists, molecular testing and immunohistochemistry assays for specific markers have evolved and gained popularity over time, in both clinical and research fields. [Extracted from the article]

The author discusses the future of histotechnology and outlines some changes in the future of histotechnology. According to the National Society of Histotechnology that histotechnology requires five basic steps on the detection of tissue abnormalities and treatment, including grossing and fixation, processing, embedding, sectioning and staining. Among other things, it notes the emergence of histotechnology in the veterinary discipline, pharmaceutical industry, and research laboratories.

Rising coal price and increasingly stringent emission policy highlight the importance of balancing economic benefit and sustainable development of coal-fired power plants. In order to make the power plant operate economically and safely, and reduce the emission of pollutants as much as possible, this paper proposed a coal blending optimization framework for coal-fired power plants. This framework constructs a set of mathematical model based on the minimization objective function, including the economy, safety and environmental protection of coal. And a multi-strategy fusion multi-objective particle swarm optimization (MSF-MOPSO) method is proposed to optimize the coal blending model. The feasibility, effectiveness and superiority of the method are theoretically verified by convergence analysis and several sets of simulation experiments. And the practical application of this framework shows that under the coal blending strategy based on different weight combinations, the algorithm proposed in this paper can produce high-quality Pareto spatial distribution. Comparing with other state-of-the-art coal blending algorithms, this method can reduce the coal purchase cost by 4.42% and ${\rm{S}}{{\rm{O}}_{\rm{2}}}$SO2 emission by 5.11% at least under the same condition. It has significant environmental protection and economic benefit. [ABSTRACT FROM AUTHOR]