Anaphylactic Shock After Cervical Conization Hemostasis With a Packing Soaked in Monsel's Solution.

Anaphylactic shock is a life-threatening medical emergency, and its successful approach depends on early recognition and treatment. We present a case report of a 54-year-old female, with the American Society of Anesthesiology (ASA) Physical Status Classification III, admitted for cervical conization. She presented with known allergies to paracetamol, diclofenac, and nimesulide, and a history of nickel contact dermatitis, with no reports of complicated anesthesia. During conization, adrenaline was infiltrated in the cervix, and hemostasis was performed with packing soaked in Monsel's solution. The immediate postoperative period in the post-anesthesia care unit was uneventful, and no drugs were administered during this period. Three hours after discharge to the ward, the patient had progressive dyspnea with desaturation and maculopapular exanthema. Anesthesia medical emergency was activated. Upon arrival of the emergency team, the patient presented: marked edema of the lips and tongue, respiratory distress, SpO2 82% (under non-rebreathing high concentration oxygen mask), audible vesicular murmur but diminished in all lung fields (without bronchospasm), blood pressure of 60/40 mmHg, increased capillary refill time (4-5 seconds), Glasgow Coma Scale score of 14, as well as generalized maculopapular exanthema and eyelid edema. Gas analysis revealed the following: pH 7.36, pO2 150, pCO2 33, HCO3 22, and lactate 2.2 mmol/L. Anaphylactic shock was immediately diagnosed without an identified causative agent. Intramuscular adrenaline (0.5mg), endovenous hydrocortisone (200 mg), clemastine (2 mg), and profuse fluid therapy were administered. There was an initial slight improvement followed by subsequent worsening. Additional administration of 0.5 mg intramuscular adrenaline and endovenous methylprednisolone (125 mg) provided similar results. Considering that no other drugs were administered in the ward, the emergency team and the attending gynecologist assumed an association between nickel allergy and the chemical composition of Monsel's solute. Thus, it was decided to remove the packing soaked in Monsel's solute from the vaginal cavity and wash it with saline solution. After removing the packing and further administration of 0.5 mg intramuscular adrenaline, there was progressive improvement in the blood pressure and SpO2. Tryptase samples collected one hour later were increased (23.9 ug/L; normal: <11.4 ug/L). The patient was shifted to the intensive care unit for surveillance, from which she was discharged after 2 days, with scheduled immunoallergology consultation, which is waiting. This case highlights the importance of causative agent identification as a key point for anaphylactic shock resolution, as well as a multidisciplinary discussion among professionals.

Pluri-IQ: Quantification of Embryonic Stem Cell Pluripotency through an Image-Based Analysis Software.

Image-based assays, such as alkaline phosphatase staining or immunocytochemistry for pluripotent markers, are common methods used in the stem cell field to assess pluripotency. Although an increased number of image-analysis approaches have been described, there is still a lack of software availability to automatically quantify pluripotency in large images after pluripotency staining. To address this need, we developed a robust and rapid image processing software, Pluri-IQ, which allows the automatic evaluation of pluripotency in large low-magnification images. Using mouse embryonic stem cells (mESC) as a model, we combined an automated segmentation algorithm with a supervised machine-learning platform to classify colonies as pluripotent, mixed, or differentiated. In addition, Pluri-IQ allows the automatic comparison between different culture conditions. This efficient user-friendly open-source software can be easily implemented in images derived from pluripotent cells or cells that express pluripotent markers (e.g., OCT4-GFP) and can be routinely used, decreasing image assessment bias.

High glucose levels affect spermatogenesis: an in vitro approach.

Besides known factors that may cause male infertility, systemic diseases such as diabetes mellitus may further exacerbate a decline in male fertility. This metabolic disease, clinically characterised by a hyperglycaemic phenotype, has devastating consequences in terms of human health, with reproductive dysfunction being one of the associated clinical complications. Nonetheless, the mechanisms responsible for such alterations are still poorly understood due to the multiplicity of factors involved in the induced pathophysiological changes. With this in mind, we focused on the main mediator of diabetes-associated alterations and performed an in vitro approach to address the effects of high glucose conditions on spermatogenesis, avoiding other confounding in vivo factors. Mouse (5 days post partum) testis fragments were cultured on agar gel stands at a gas-liquid interface with either 5, 25 or 50mM D-glucose for 3 weeks. Stereological analysis revealed that high D-glucose levels increased Sertoli cell number (P<0.05) and decreased tubular luminal area (P<0.01), suggesting an impairment of this somatic cell type. Moreover, higher proliferative activity in a TM4 Sertoli cell line exposed to high D-glucose was found (P<0.05) without compromising cell viability (P>0.05), further suggesting altered Sertoli cell maturation. Overall, high D-glucose concentrations may lead to impairment of Sertoli cell function, which, given their significant role in spermatogenic control, may compromise male fertility.

Sirtuins in metabolism, stemness and differentiation.

BACKGROUND: Pluripotent stem cells promise innovative approaches for enduring diseases, including disease modeling and drug screens. Accordingly, efforts have been undertaken in order to efficiently reprogram somatic cells to pluripotency, and then differentiate them into pure cultures of specific cell lineages. However, the latter step remains mostly elusive, and, in order to better control differentiation and design more efficient differentiation strategies, the cellular mechanisms behind different pluripotency stages that mimic embryonic development are being actively addressed. SCOPE OF REVIEW: Metabolism is one of many cellular processes that are in constant adjustment during mammalian embryo development, as well as in pluripotent stem cell establishment and differentiation. Thus, the role of molecular pathways known to be involved in metabolic control has been recently addressed as potential modulators of pluripotency. Notably, mammalian sirtuins have emerged as master regulators of many cellular processes, including epigenetics and metabolism. In this review we address the potential developmental role of sirtuins, with a particular focus on sirtuin 1. MAJOR CONCLUSIONS: This review focuses on the most recent studies implying sirtuins as regulators of pluripotency and differentiation of pluripotent stem cells, highlighting metabolic control as associated with the control of pluripotency. It notably stresses the role of sirtuin 1 in these processes, creating parallels between in vitro manipulations and developmental cues. GENERAL SIGNIFICANCE: Using metabolic control in order to determine cellular fate, both in terms of somatic cell reprogramming to pluripotency and pluripotent stem cell differentiation, is a topic of increasing interest, and sirtuins are key players in these efforts.

Data on the potential impact of food supplements on the growth of mouse embryonic stem cells.

The use of new compounds as dietary supplements is increasing, but little is known in terms of possible consequences of their use. Pluripotent stem cells are a promising research tool for citotoxicological research for evaluation of proliferation, cell death, pluripotency and differentiation. Using the mouse embryonic stem cell (mESC) model, we present data on three different compounds that have been proposed as new potential supplements for co-adjuvant disease treatments: kaempferol, berberine and Tauroursodeoxycholic acid (TUDCA). Cell number and viability were monitored following treatment with increased concentrations of each drug in pluripotent culture conditions.

Different concentrations of kaempferol distinctly modulate murine embryonic stem cell function.

Kaempferol (3,4',5,7-tetrahydroxyflavone) is a natural flavonoid with several beneficial and protective effects. It has been demonstrated that kaempferol has anticancer properties, particularly due to its effects on proliferation, apoptosis and the cell cycle. However, possible effects on pluripotent embryonic stem cell function have not yet been addressed. Embryonic stem cells have the ability to self-renew and to differentiate into all three germ layers with potential applications in regenerative medicine and in vitro toxicology. We show that exposure of murine embryonic stem cells (mESC) to high concentrations of kaempferol (200 µM) leads to decreased cell numbers, although the resulting smaller cell colonies remain pluripotent. However, lower concentrations of this compound (20 µM) increase the expression of pluripotency markers in mESCs. Mitochondrial membrane potential and mitochondrial mass are not affected, but a dose-dependent increase in apoptosis takes place. Moreover, mESC differentiation is impaired by kaempferol, which was not related to apoptosis induction. Our results show that low concentrations of kaempferol can be beneficial for pluripotency, but inhibit proper differentiation of mESCs. Additionally, high concentrations induce apoptosis and increase mitochondrial reactive oxygen species (ROS).