Metallothioneins (MTs) are small molecular weight stress response proteins that play a central role as reservoir of essential divalent heavy metal cations such as zinc and copper, and also can diminish the effects of toxic heavy metals such as mercury and cadmium. Historically, MT has been considered to be an intracellular protein with roles to play in the management of heavy metals, as a regulator of cellular redox potential, and as a buffer of free radicals. Our recent studies have highlighted immunomodulatory role of MT in inflammatory diseases and also in the progression of metastatic cell movement. Hence, manipulation and detection of MT is essential for its possible use as a diagnostic and in therapeutic interventions of chronic inflammation. This review describes procedures used to detect MT using techniques such as western immunoblot, competition ELISA, flow cytometry and immunohistochemistry. Additionally, it also describes the use of a colorimetric cell proliferation assay (CellTiter 96 AQ One Solution/MTS) to study the proliferative effect of MT. © 2017 by John Wiley & Sons, Inc.

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Drug-induced gingival overgrowth is an adverse effect of certain drugs, including amlodipine, in genetically susceptible individuals. Although the exact mechanism of gingival hypertrophy remains unclear, a unifying multifactorial hypothesis has been constructed. Gingival hypertrophy causes difficulty in speech and mastication, poor oral hygiene, and poor aesthetic appearance. Here, we present the case of a 49-year-old woman who developed gum hypertrophy following amlodipine use for two years. Maintenance of oral hygiene and substitution of offending agent is commonly the first step in management.

To sustain energy-demanding developmental processes, oocytes must accumulate adequate stores of metabolic substrates and mitochondrial numbers prior to the initiation of maturation. In the past, researchers have utilized pooled samples to study oocyte metabolism, and studies that related multiple metabolic outcomes in single oocytes, such as ATP concentration and mitochondrial DNA copy number, were not possible. Such scenarios decreased sensitivity to intraoocyte metabolic relationships and made it difficult to obtain adequate sample numbers during studies with limited oocyte availability. Therefore, we developed and validated procedures to measure both mitochondrial DNA (mtDNA) copy number and ATP quantity in single oocytes. Validation of our procedures revealed that we could successfully divide oocyte lysates into quarters and measure consistent results from each of the aliquots for both ATP and mtDNA copy number. Coefficient of variation between the values retrieved for mtDNA copy number and ATP quantity quadruplicates were 4.72 ± 0.98 and 1.61 ± 1.19, respectively. We then utilized our methodology to concurrently measure mtDNA copy number and ATP quantity in germinal vesicle (GV) and metaphase two (MII) stage oocytes. Our methods revealed a significant increase in ATP levels (GV = 628.02 ± 199.53 pg, MII = 1326.24 ± 199.86 pg, p < 0.001) and mtDNA copy number (GV = 490,799.4 ± 544,745.9 copies, MII = 1,087,126.9 ± 902,202.8 copies, p = 0.035) in MII compared to GV stage oocytes. This finding is consistent with published literature and provides further validation of the accuracy of our methods. The ability to produce consistent readings and expected results from aliquots of the lysate from a single oocyte reveals the sensitivity and feasibility of using this method.

Dysregulated cell movement can lead to developmental abnormalities, neoplasia, and immune system disorders, and there are a variety of contexts in which xenobiotics (and biologic) effects on this movement are of interest. Many toxins and toxicants have been shown to disrupt controlled cell movement. Identification of compounds that affect cell movement is crucial to drug discovery. Drug components may have unexpected consequences with respect to cell motility, which would exclude these compounds in drug development. Finally, the development of drugs that target chemotactic pathways may be useful in the treatment of tumors, which often reprogram chemotactic pathways to become metastatic. The effects of these agents on cell movement can be measured using several different in vitro chemotactic assays. This review details the procedures of three in vitro measurements of chemotaxis: the Boyden chamber, the under-agarose assay, and the automated, real-time, ECIS/Taxis assay, and discusses the inferences that can be drawn from the results of such studies.

Systemic rheumatoid diseases (SRDs) are autoimmune and inflammatory disorders that affect multiple organ systems, impacting patients’ quality of life, and survival rates. Standard treatment requires continuous drug therapy and immunosuppression. Chimeric antigen receptor (CAR) T cell therapy has the potential to target and eliminate pathologically activated immune cells and re-establish tolerance in organs affected by dysregulated immunity, making them a promising treatment option for autoimmune diseases. In autoimmune diseases, CAR T cells have the advantage of being able to kill B cells effectively without the need for an accessory cell type. Additionally, CAR T cells targeting CD19 have shown promise in comprehensive B cell aplasia, preserving pre-existing humoral immunity, and specifically eliminating pathogenic B cells. CAR T cell therapy’s limited use in SRDs is due to its inability to effectively target the various autoreactive lymphocytes present. Researchers are developing a universal CAR T cell therapy that detects and targets autoreactive lymphocytes using major epitope peptides, though further studies are required. Moreover, adoptive transfer of CAR-Tregs has shown promise for effectively reducing inflammation and treating autoimmunity. Through this exploration, the authors hope to provide a comprehensive understanding of the current state of research on this topic, identify areas for further study, and promote the advancement of CAR T cell therapy as a treatment option for SRDs.

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Scrub typhus is an arthropod-borne fever that follows the bite of the larval form of Leptotrombidium mite carrying Orientia tsutsugamushi. It remains a serious health problem in the Asia-Pacific region. While it commonly presents as an undifferentiated fever with chills and an eschar, complications like pneumonitis, acute respiratory distress syndrome, disseminated intravascular coagulation, and meningoencephalitis may cause scrub typhus to be fatal. However, regardless of the dramatic presentation, treatment with antibiotics, preferably doxycycline or even azithromycin, is effective in recovery.In this case report, we present a case of meningitis and cerebellar involvement in an adolescent with positive scrub typhus serology in the absence of an eschar. This brought forward a diagnostic delay as other infections including tuberculosis were considered before scrub typhus due to unusual presenting symptoms and the lack of an eschar. Thus, in cases like these, it becomes imperative to be aware of the unusual manifestations to initiate antibiotics on time and prevent further complications.