Ashwagandha-Induced Programmed Cell Death in the Treatment of Breast Cancer.

The aim of this review is to provide experimental evidence for the programmed-death activity of Ashwagandha (Withania somnifera) in the anti-cancer therapy of breast cancer. The literature search was conducted using online electronic databases (Google Scholar, PubMed, Scopus). Collection schedule data for the review article covered the years 2004-2024. Ashwagandha active substances, especially Withaferin A (WA), are the most promising anti-cancer compounds. WS exerts its effect on breast cancer cells by inducing programmed cell death, especially apoptosis, at the molecular level. Ashwagandha has been found to possess a potential for treating breast cancer, especially estrogen receptor/progesterone receptor (ER/PR)-positive and triple-negative breast cancer.

Immunodominant AH1 Antigen-Deficient Necroptotic, but Not Apoptotic, Murine Cancer Cells Induce Antitumor Protection.

Immunogenic cell death (ICD) occurs when a dying cell releases cytokines and damage-associated molecular patterns, acting as adjuvants, and expresses Ags that induce a specific antitumor immune response. ICD is studied mainly in the context of regulated cell death pathways, especially caspase-mediated apoptosis marked by endoplasmic reticulum stress and calreticulin exposure and, more recently, also in relation to receptor-interacting protein kinase-driven necroptosis, whereas unregulated cell death like accidental necrosis is nonimmunogenic. Importantly, the murine cancer cell lines used in ICD studies often express virally derived peptides that are recognized by the immune system as tumor-associated Ags. However, it is unknown how different cell death pathways may affect neoepitope cross-presentation and Ag recognition of cancer cells. We used a prophylactic tumor vaccination model and observed that both apoptotic and necroptotic colon carcinoma CT26 cells efficiently immunized mice against challenge with a breast cancer cell line that expresses the same immunodominant tumor Ag, AH1, but only necroptotic CT26 cells would mount an immune response against CT26-specific neoepitopes. By CRISPR/Cas9 genome editing, we knocked out AH1 and saw that only necroptotic CT26 cells were still able to protect mice against tumor challenge. Hence, in this study, we show that endogenous AH1 tumor Ag expression can mask the strength of immunogenicity induced by different cell death pathways and that upon knockout of AH1, necroptosis was more immunogenic than apoptosis in a prophylactic tumor vaccination model. This work highlights necroptosis as a possible preferred ICD form over apoptosis in the treatment of cancer.

Vaccination with Necroptotic Cancer Cells Induces Efficient Anti-tumor Immunity.

Successful immunogenic apoptosis in experimental cancer therapy depends on the induction of strong host anti-tumor responses. Given that tumors are often resistant to apoptosis, it is important to identify alternative molecular mechanisms that elicit immunogenic cell death. We have developed a genetic model in which direct dimerization of FADD combined with inducible expression of RIPK3 promotes necroptosis. We report that necroptotic cancer cells release damage-associated molecular patterns and promote maturation of dendritic cells, the cross-priming of cytotoxic T cells, and the production of IFN-γ in response to tumor antigen stimulation. Using both FADD-dependent and FADD-independent RIPK3 induction systems, we demonstrate the efficient vaccination potential of immunogenic necroptotic cells. Our study broadens the current concept of immunogenic cell death and opens doors for the development of new strategies in cancer therapy.

Bone Mineral Density and Biochemical Markers of Bone Metabolism in Women Engaging in Recreational Horseback Riding.

BACKGROUND: An increased occurrence of lifestyle-related diseases such as osteoporosis indicates the necessity for taking preventive action, including regularly engaging in physical activity. The aim of the study was to assess the areal bone mineral density (aBMD) and bone turnover markers levels in young adult women engaging in recreational horseback riding and to determine the relationship between training characteristics and bone metabolism indices. METHODS: The study involved 43 women: 23 equestrians and 20 age- and body mass index-matched controls. The hip and spine aBMD and serum levels of the bone turnover markers: osteocalcin and collagen type I cross-linked C-telopeptide were measured. RESULTS: No significant differences were found in somatic features, concentrations of bone turnover markers, or bone mass variables. Correlation analysis of the equestrian participants showed significant relationship between body mass and BMDL1-L4 (P < .05) as well as between BMI and BMDL1-L4 (P ≤ .01) and z-score L1-L4 (P < .05). CONCLUSIONS: The study showed no differences in bone mass and levels of bone metabolic indices between groups of women practicing horseback riding at the recreational level and subjects who do not participate in frequent systematic physical activity. No relationship between training characteristics and bone turnover markers were found.

Immunogenic cell death and DAMPs in cancer therapy.

Although it was thought that apoptotic cells, when rapidly phagocytosed, underwent a silent death that did not trigger an immune response, in recent years a new concept of immunogenic cell death (ICD) has emerged. The immunogenic characteristics of ICD are mainly mediated by damage-associated molecular patterns (DAMPs), which include surface-exposed calreticulin (CRT), secreted ATP and released high mobility group protein B1 (HMGB1). Most DAMPs can be recognized by pattern recognition receptors (PRRs). In this Review, we discuss the role of endoplasmic reticulum (ER) stress and reactive oxygen species (ROS) in regulating the immunogenicity of dying cancer cells and the effect of therapy-resistant cancer microevolution on ICD.

ER stress-induced inflammation: does it aid or impede disease progression?

Different lines of research have revealed that pathways activated by the endoplasmic reticulum (ER) stress response induce sterile inflammation. When activated, all three sensors of the unfolded protein response (UPR), PERK, IRE1, and ATF6, participate in upregulating inflammatory processes. ER stress in various cells plays an important role in the pathogenesis of several diseases, including obesity, type 2 diabetes, cancer, and intestinal bowel and airway diseases. Moreover, it has been suggested that ER stress-induced inflammation contributes substantially to disease progression. However, this generalization can be challenged at least in the case of cancer. In this review, we emphasize that ER stress can either aid or impede disease progression via inflammatory pathways depending on the cell type, disease stage, and type of ER stressor.

ATP release from dying autophagic cells and their phagocytosis are crucial for inflammasome activation in macrophages.

Pathogen-activated and damage-associated molecular patterns activate the inflammasome in macrophages. We report that mouse macrophages release IL-1ß while co-incubated with pro-B (Ba/F3) cells dying, as a result of IL-3 withdrawal, by apoptosis with autophagy, but not when they are co-incubated with living, apoptotic, necrotic or necrostatin-1 treated cells. NALP3-deficient macrophages display reduced IL-1ß secretion, which is also inhibited in macrophages deficient in caspase-1 or pre-treated with its inhibitor. This finding demonstrates that the inflammasome is activated during phagocytosis of dying autophagic cells. We show that activation of NALP3 depends on phagocytosis of dying cells, ATP release through pannexin-1 channels of dying autophagic cells, P(2)X(7) purinergic receptor activation, and on consequent potassium efflux. Dying autophagic Ba/F3 cells injected intraperitoneally in mice recruit neutrophils and thereby induce acute inflammation. These findings demonstrate that NALP3 performs key upstream functions in inflammasome activation in mouse macrophages engulfing dying autophagic cells, and that these functions lead to pro-inflammatory responses.

A novel pathway combining calreticulin exposure and ATP secretion in immunogenic cancer cell death.

Surface-exposed calreticulin (ecto-CRT) and secreted ATP are crucial damage-associated molecular patterns (DAMPs) for immunogenic apoptosis. Inducers of immunogenic apoptosis rely on an endoplasmic reticulum (ER)-based (reactive oxygen species (ROS)-regulated) pathway for ecto-CRT induction, but the ATP secretion pathway is unknown. We found that after photodynamic therapy (PDT), which generates ROS-mediated ER stress, dying cancer cells undergo immunogenic apoptosis characterized by phenotypic maturation (CD80(high), CD83(high), CD86(high), MHC-II(high)) and functional stimulation (NO(high), IL-10(absent), IL-1ß(high)) of dendritic cells as well as induction of a protective antitumour immune response. Intriguingly, early after PDT the cancer cells displayed ecto-CRT and secreted ATP before exhibiting biochemical signatures of apoptosis, through overlapping PERK-orchestrated pathways that require a functional secretory pathway and phosphoinositide 3-kinase (PI3K)-mediated plasma membrane/extracellular trafficking. Interestingly, eIF2α phosphorylation and caspase-8 signalling are dispensable for this ecto-CRT exposure. We also identified LRP1/CD91 as the surface docking site for ecto-CRT and found that depletion of PERK, PI3K p110α and LRP1 but not caspase-8 reduced the immunogenicity of the cancer cells. These results unravel a novel PERK-dependent subroutine for the early and simultaneous emission of two critical DAMPs following ROS-mediated ER stress.

Severity of doxorubicin-induced small intestinal mucositis is regulated by the TLR-2 and TLR-9 pathways.

Intestinal mucositis is a serious complication of cancer chemotherapy and radiotherapy; it frequently compromises treatment and dramatically reduces the quality of life of patients. Different approaches to limit the damage to the intestine during anti-cancer therapy have been largely ineffective due to insufficient knowledge of the mechanism of mucositis development. This study aimed to define the role of TLR-2 and TLR-9 in the modulation of small intestinal damage in a model of doxorubicin-induced mucositis. Doxorubicin-induced intestinal damage was verified by a histological score (HS), analysis of leukocyte influx into the lamina propria, and determination of the number of apoptotic cells. Additionally, the activation status of glycogen synthase kinase 3ß (GSK-3ß) was assessed. Wild-type (WT) mice injected with doxorubicin demonstrated severe intestinal damage (HS 8.0 ± 0.81), reduction of villus length to 43.9% ± 13.7% of original length, and increased influx of leukocytes as compared to vehicle-injected mice (HS 1.33 ± 1.15). The protective effect of TLR-2 or TLR-9 deficiency was associated with a significant decrease of the HS as compared to WT mice. In the ileum, a minor reduction of villus length and a decreased number of infiltrating leukocytes and TUNEL-positive cells was observed. We demonstrate that the TLR-9 antagonist ODN2088 reduces doxorubicin-induced intestinal damage. Furthermore, we show that GSK-3ß activity is inhibited in the absence of TLR-2. The protective capacity of GSK-3ß suppression was observed in WT mice by inhibiting it with the specific inhibitor SB216763. Overall, our findings demonstrate that the TLR-2/GSK-3ß and TLR-9 signalling pathways play a central role in the development of intestinal mucositis and we suggest a new therapeutic strategy for limiting doxorubicin-induced intestinal inflammation.

Detection of Helicobacter pylori and cagA gene in nasal polyps and benign laryngeal diseases.

BACKGROUND AND AIMS: Helicobacter pylori is the most common etiological factor of chronic infection worldwide. It has also been found in human dental plaques, mouth, saliva, tonsils and adenoid tissue, medial ear or nasal polyps and sinuses mucosa, as well in several benign and malignant lesions of the larynx and pharynx. The aim of the study was to investigate the association of H. pylori colonization in chronic rhinosinusitis and benign laryngeal diseases. METHODS: The prospective, controlled study involved a series of 30 patients with nasal polyps and normal nasal mucosa and 30 patients with benign laryngeal diseases. Samples of 10-15 mg obtained from fresh tissues were used for nucleic acid purification. All samples were subjected to H. pylori ureA detection by the PCR H. pylori diagnostic test. Samples that were positive for ureA H. pylori gene were evaluated for cagA H. pylori gene. RESULTS: H. pylori DNA (ureA gene) was detected in all patients with nasal polyps, concha bullosa and laryngeal diseases. Presence of H. pylori cagA gene was identified in 7 (23.3%) of 30 patients of H. pylori-positive larynx samples and no positive result was observed in nasal polyps and concha bullosa. CONCLUSIONS: Our results reveal the presence of H. pylori DNA in nasal polyps, concha bullosa and benign larynx diseases. cagA-positive H. pylori was observed only in laryngeal tissues. These results may have implications for a possible role of H. pylori in laryngeal diseases.

[Urinary system as a primary source of bloodstream infection]. / Uklad moczowy jako pierwotne zródlo zakazenia krwi.

The aim of this study was the determinantion of the appearance of urosepsis in patients in the Department of Nephrology Hypertension and internal Diseases, Dialysis Unit, Department of General, Oncologic and Pediatric Urology and Department of Transplantology and Surgery of University Hospital nr 1 of dr. A. Jurasza in Bydgoszcz. In the period of 2006-2008, total 233 isolates were obtained from positive blood culture and 28.8% of these was also related to urinary tract infection. The majority of cases of urosepsis was observed in urologic and transplantologic wards. Among investigated strains, the most frequency isolated species was E. coli. All Enterobacteriaceae strains examined in our study were susceptible to imipenem, the highest rate of resistance was found for ampicillin, six isolates was producing extended-spectrum beta-lactamases and four-AmpC beta-lactamases. Among P. aeruginosa strains there was no resistance to netilmicin and one or two isolates was producing AmpC beta-lactamases or ES L, respectively.

Measurement of coronary fractional flow reserve. Standard doses of intracoronary adenosine are insufficient to induce maximal hyperaemia.

BACKGROUND: Induction of maximal hyperaemia is a prerequisite for a reliable estimation of fractional flow reserve (FFR) in a moderate coronary artery stenosis. Intracoronary adenosine is the most frequently used agent to achieve maximal hyperaemia. However, an insufficient dose of adenosine may induce only partial hyperaemia, thus artificially increasing the FFR values. AIM: To assess the tolerability and effects on FFR value of increased doses of adenosine. METHODS: FFR was measured in 36 patients with 53 moderate coronary lesions. In order to induce maximal hyperaemia and assess FFR in the targeted coronary artery, intracoronary adenosine in a dose of 30 micro g was administered twice (FFR30). Next, 60 micro g of adenosine was tested twice (FFR60). In addition, in some patients with left coronary artery stenosis, 90 micro g of adenosine was also injected (FFR90). RESULTS: No significant side effects were noted except a transient, self-terminating episode of a second degree atrio-ventricular block in one patient. The mean value of FFR30 was significantly higher than FFR60 (0.854+/-0.152 vs 0.836+/-0.162, p<0.001), and the mean difference between these two measurements was 0.018+/-0.036. In 29 (54.7%) evaluated lesions, FFR30 values were higher than FFR60; in 12 (22.6%) measurements the difference exceeded 0.02, and in 8 (15%) cases - 0.05. The use of 90 micro g of adenosine did not further decrease FFR in any of the cases. CONCLUSIONS: An increase of the adenosine dose from 30 micro g to 60 micro g was well tolerated and caused further decrease in the FFR values which may be of clinical importance in some patients. The use of 90 micro g of adenosine did not further decrease FFR.