Acute toxicity and antitumor potential of 1,3,4-trisubstituted-1,2,3-triazole dhmtAc-loaded liposomes on a triple-negative breast cancer model.

For the first time, compounds developed from the 1,2,3-triazole scaffold were evaluated as novel drugs to treat triple-negative breast cancer (TNBC). Four organic salts were idealized as nonclassical bioisosteres of miltefosine, which is used in the topical treatment for skin metastasizing breast carcinoma. Among them, derivative dhmtAc displayed better solubility and higher cytotoxicity against the human breast adenocarcinoma cell line and mouse 4T1 cell lines, which are representatives of TNBC. In vitro assays revealed that dhmtAc interferes with cell integrity, confirmed by lactate dehydogenase leakage. Due to its human peripheral blood mononuclear cell (PBMC) toxicity, dhmtAc in vivo studies were carried out with the drug incorporated in a long-circulating and pH-sensitive liposome (SpHL-dhmtAc), and the acute toxicity in BALB/c mice was determined. Free dhmtAc displayed cardiac and pulmonary toxicity after the systemic administration of 5 mg/kg doses. On the other hand, SpHL-dhmtAc displayed no toxicity at 20 mg/kg. The in vivo antitumor effect of SpHL-dhmtAc was investigated using the 4T1 heterotopic murine model. Intravenous administration of SpHL-dhmtAc reduced the tumor volume and weight, without interfering with the body weight, compared with the control group and the dhmtAc free form. The incorporation of the triazole compound in the liposome allowed the demonstration of its anticancer potential. These findings evidenced 1,3,4-trisubstituted-1,2,3-triazole as a promising scaffold for the development of novel drugs with applicability for the treatment of patients with TNBC.

Molecular Study of Torque Teno Virus in Patients with Acute Gastroenteritis.

BACKGROUND: Torque teno virus (TTV) is a single stranded non enveloped DNA virus. Various studies have found a high prevalence of TTV in different populations and in different human samples including blood and stool. OBJECTIVE: The aim of the present study was to evaluate the prevalence of TTV in adult patients with acute gastroenteritis in stool samples by semi-nested polymerase chain reaction (PCR). METHODS: This study was a retrospective, cross-sectional study carried out on 100 preserved stool samples from adult patients with simple community acquired diarrhea without dehydration. Stool samples were subjected to antigen detection of rotavirus and norovirus by enzyme linked immunosorbent assay (ELISA). Detection of TTV was performed by the use of semi- nested PCR. RESULTS: The detected viruses were TTV by semi-nested PCR in 83% of the patients, followed by both norovirus and rotavirus in 20% of patients each. TTV was present without any other studied virus in 52% of the samples, the norovirus antigen was detected as a single virus in 2%, and rotavirus was detected as a single virus in 3%. No viruses were detected in 11% of the stool samples. Norovirus was associated with TTV in 17 isolates and as a sole virus in three samples (p = 0.5). Rotavirus was associated with TTV in 17 isolates and alone in three. CONCLUSIONS: The data of the present study show a high prevalence of TTV in stool samples from adults with acute gastroenteritis. The presence of rotavirus and norovirus was also a common finding in these patients. There were no detected effects on the clinical features of gastroenteritis associated with the presence of TTV in acute gastroenteritis.

The synthetic peptide LyeTxI-b derived from Lycosa erythrognatha spider venom is cytotoxic to U-87 MG glioblastoma cells.

Antimicrobial peptides present a broad spectrum of therapeutic applications, including their use as anticancer peptides. These peptides have as target microbial, normal, and cancerous cells. The oncological properties of these peptides may occur by membranolytic mechanisms or non-membranolytics. In this work, we demonstrate for the first time the cytotoxic effects of the cationic alpha-helical antimicrobial peptide LyeTx I-b on glioblastoma lineage U87-MG. The anticancer property of this peptide was associated with a membranolytic mechanism. Loss of membrane integrity occurred after incubation with the peptide for 15 min, as shown by trypan blue uptake, reduction of calcein-AM conversion, and LDH release. Morphological studies using scanning electron microscopy demonstrated disruption of the plasma membrane from cells treated with LyeTx I-b, including the formation of holes or pores. Transmission electron microscopy analyses showed swollen nuclei with mild DNA condensation, cell volume increase with an electron-lucent cytoplasm and organelle vacuolization, but without the rupture of nuclear or plasmatic membranes. Morphometric analyses revealed a high percentage of cells in necroptosis stages, followed by necrosis and apoptosis at lower levels. Necrostatin-1, a known inhibitor of necroptosis, partially protected the cells from the toxicity of the peptide in a concentration-dependent manner. Imaging flow cytometry confirmed that 59% of the cells underwent necroptosis after 3-h incubation with the peptide. It is noteworthy that LyeTx I-b showed only mild cytotoxicity against normal fibroblasts of human and monkey cell lines and low hemolytic activity in human erythrocytes. All data together point out the anticancer potential of this peptide.

Assessment of long noncoding RNA CCAT1 using real time-polymerase chain reaction in colorectal cancer patients.

Colorectal cancer (CRC) is linked to high mortality, mainly when discovered in its advanced stages. Several studies have pointed to the role of epigenetic factors in CRC and other cancers. Long non-coding RNAs (lncRNAs) are involved in the initiation, progression, metastasis, and modulation of the response to chemotherapeutic modalities of CRC as vital contributors to epigenetic mechanisms. Colon cancer-associated transcript-1 (CCAT1) is one of the lncRNAs that have been dysregulated in serum samples, providing a non-invasive route for diagnosing CRC patients. This study aimed to determine the role of CCAT1 expression as diagnostic and prognostic markers. We tested the associations of CCAT1 expression with serum carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA 19-9). The study included three groups: 41 patients with colorectal cancer, 39 patients with precancerous benign colorectal diseases, and 20 normal control individuals. CEA and CA 19-9 were measured by an immunoassay automated system. The expression level of CCAT1 was assessed by a real-time polymerase chain reaction. There was a statistically significant elevation of serum CEA levels in patients with CRC compared to patients with precancerous benign colorectal diseases. Furthermore, there was no statistically significant difference in serum CA 19-9 levels between all groups (p = 0.102). Interestingly, CCAT1 expression was significantly upregulated in the blood of CRC patients compared to the precancerous benign colorectal diseases group (p = 0.009) and the control group (p <0.001). Also, expression of CCAT1 was significantly elevated in patients with precancerous benign colorectal diseases compared to the control group (p=0.004). In conclusion, measuring the expression level of CCAT1 is more advised than assessment of CEA and CA 19-9 for the early diagnosis and prognosis of colorectal cancer.

Interactomic exploration of LRRC8A in volume-regulated anion channels.

Ion channels are critical in enabling ion movement into and within cells and are important targets for pharmacological interventions in different human diseases. In addition to their ion transport abilities, ion channels interact with signalling and scaffolding proteins, which affects their function, cellular positioning, and links to intracellular signalling pathways. The study of "channelosomes" within cells has the potential to uncover their involvement in human diseases, although this field of research is still emerging. LRRC8A is the gene that encodes a crucial protein involved in the formation of volume-regulated anion channels (VRACs). Some studies suggest that LRRC8A could be a valuable prognostic tool in different types of cancer, serving as a biomarker for predicting patients' outcomes. LRRC8A expression levels might be linked to tumour progression, metastasis, and treatment response, although its implications in different cancer types can be varied. Here, publicly accessible databases of cancer patients were systematically analysed to determine if a correlation between VRAC channel expression and survival rate exists across distinct cancer types. Moreover, we re-evaluated the impact of LRRC8A on cellular proliferation and migration in colon cancer via HCT116 LRRC8A-KO cells, which is a current topic of debate in the literature. In addition, to investigate the role of LRRC8A in cellular signalling, we conducted biotin proximity-dependent identification (BioID) analysis, revealing a correlation between VRAC channels and cell-cell junctions, mechanisms that govern cellular calcium homeostasis, kinases, and GTPase signalling. Overall, this dataset improves our understanding of LRRC8A/VRAC and explores new research avenues while identifying promising therapeutic targets and promoting inventive methods for disease treatment.

Simultaneous targeting of mitochondrial Kv1.3 and lysosomal acid sphingomyelinase amplifies killing of pancreatic ductal adenocarcinoma cells in vitro and in vivo.

Pancreas ductal adenocarcinoma (PDAC) remains a malignant tumor with very poor prognosis and low 5-year overall survival. Here, we aimed to simultaneously target mitochondria and lysosomes as a new treatment paradigm of malignant pancreas cancer in vitro and in vivo. We demonstrate that the clinically used sphingosine analog FTY-720 together with PAPTP, an inhibitor of mitochondrial Kv1.3, induce death of pancreas cancer cells in vitro and in vivo. The combination of both drugs results in a marked inhibition of the acid sphingomyelinase and accumulation of cellular sphingomyelin in vitro and in vivo in orthotopic and flank pancreas cancers. Mechanistically, PAPTP and FTY-720 cause a disruption of both mitochondria and lysosomes, an alteration of mitochondrial bioenergetics and accumulation of cytoplasmic Ca2+, events that collectively mediate cell death. Our findings point to an unexpected cross-talk between lysosomes and mitochondria mediated by sphingolipid metabolism. We show that the combination of PAPTP and FTY-720 induces massive death of pancreas cancer cells, thereby leading to a substantially delayed and reduced PDAC growth in vivo. KEY MESSAGES: FTY-720 inhibits acid sphingomyelinase in pancreas cancer cells (PDAC). FTY-720 induces sphingomyelin accumulation and lysosomal dysfunction. The mitochondrial Kv1.3 inhibitor PAPTP disrupts mitochondrial functions. PAPTP and FTY-720 synergistically kill PDAC in vitro. The combination of FTY-720 and PAPTP greatly delays PDAC growth in vivo.

Study of Plasmid-Mediated Quinolone Resistance in Escherichia coli from Nosocomial Urinary Tract Infections.

OBJECTIVE: The aim of the present study was to study the prevalence of plasmid-mediated quinolone resistance (PMQR) genes (qnrA, qnrB, qnrC, qnrD, qnrS, qepA, oqxA, oqxB and aac) in Escherichia coli (E. coli) isolated from patients with nosocomial urinary tract infections (UTIs) and its relation to the extended-spectrum ß-lactamase (ESBL) production. ; Methods: A cross-sectional study was carried out on 200 non-duplicated isolates of E. coli isolated from patients with nosocomial UTIs. E.coli isolates were subjected to antibiotic susceptibility testing by disc diffusion method, determination of minimum inhibitory concentrations (MICs) of ciprofloxacin by Epsillometer (E) test strips, detection of ESBL production by double disc synergy method and detection of qnrA, qnrB, qnrC, qnrD, qnrS, qepA, oqxA, oqxB and aac genes by polymerase chain reaction (PCR). ; Results: The antimicrobial susceptibility testing of the isolated E. coli revealed a high frequency of resistance to ampicillin (73.5%), ceftazidime (72%) and imipenem (71.5%). The less frequent resistance was for aztreonam (21.5%), amikacin (36.5%) and gentamicin (38.5%). ESBL production was found in 131 isolates (65.5%) and phenotypic quinolone resistance was detected by MIC in 65 isolates (32.5%), with 52.3% of them showed high resistance to ciprofloxacin with an MIC more than 32 µg/ml. PMQR genes were found in 40 isolates. The frequency of the detected genes was 40%, 37.5%, 35%, 20% and 5% for qnrA, qnrS, qepA, qnrB and oqxA, respectively. Significant association was found between the presence of PMQR genes and ESBL production (P=0.0001). ; Conclusion: The study highlights the prevalence of PMQR genes in E. coli with high association with the ESBL phenotype. This finding is a sign of limited therapeutic options for E. coli.

Incidence and Outcomes of Acute Kidney Injury in Critically Ill Patients with Coronavirus Disease 2019.

Patients with more severe cases of coronavirus disease-19 (COVID-19) may be at greater risk for developing acute kidney injury (AKI). The aim of our study was to analyze incidence and outcomes of AKI in critically ill patients with COVID-19. Our study prospectively followed about 198 patients with COVID-19 admitted to intensive care unit (ICU), Al Adan Hospital, Kuwait, for developing AKI and outcomes. Age, gender, nationality, history of hypertension, diabetes mellitus, ischemic heart disease, congestive heart failure, bronchial asthma, and chronic obstructive pulmonary disease were analyzed. The need for mechanical ventilation (MV), extracorporeal membrane oxygenation, inotropes, and medications was recorded. Causes of AKI, indication of dialysis, dialysis modality, dialysis outcomes, and mortality were analyzed. Our study reported that61 out of 198 (30.8%) ICU patients positive for COVID-19, developed AKI according to the Kidney Disease Improving Global Outcomes definition of AKI. Forty-eight out of 61 (79%) patients need continuous renal replacement therapy using continuous venovenous hemodiafiltration. Thirty-seven (61%) out of 61 patients were with severe sepsis syndrome. The most common cause of AKI was sepsis, cytokine storm, hypovolemia, heart failure, MV, and nephrotoxic drugs. Twenty-four patients (39%) out of 61 patients died, and the most common cause of death was sepsis, cytokine storm with respiratory failure, heart failure, and AKI. The outcome of AKI was as follows: six patients (10%) had complete recovery, five patients had partial recovery (8%), and 26 (43%) patients became dialysis dependent. Incidence of AKI is high in ICU COVID-19 patients and is associated with poor outcomes and high mortality. Early detection and specific therapy of kidney changes, including adequate hemodynamic support and avoidance of nephrotoxic drugs, may help to improve critically ill patients with COVID-19.

LyeTx I-b Peptide Attenuates Tumor Burden and Metastasis in a Mouse 4T1 Breast Cancer Model.

Cationic anticancer peptides have exhibited potent anti-proliferative and anti-inflammatory effects in neoplastic illness conditions. LyeTx I-b is a synthetic peptide derived from Lycosa erythrognatha spider venom that previously showed antibiotic activity in vitro and in vivo. This study focused on the effects of LyeTxI-b on a 4T1 mouse mammary carcinoma model. Mice with a palpable tumor in the left flank were subcutaneously or intratumorally injected with LyeTx I-b (5 mg/kg), which significantly decreased the tumor volume and metastatic nodules. Histological analyses showed a large necrotic area in treated primary tumors compared to the control. LyeTxI-b reduced tumor growth and lung metastasis in the 4T1 mouse mammary carcinoma model with no signs of toxicity in healthy or cancerous mice. The mechanism of action of LyeTx I-b on the 4T1 mouse mammary carcinoma model was evaluated in vitro and is associated with induction of apoptosis and cell proliferation inhibition. Furthermore, LyeTx I-b seems to be an efficient regulator of the 4T1 tumor microenvironment by modulating several cytokines, such as TGF-ß, TNF-α, IL-1ß, IL-6, and IL-10, in primary tumor and lung, spleen, and brain. LyeTx I-b also plays a role in leukocytes rolling and adhesion into spinal cord microcirculation and in the number of circulating leukocytes. These data suggest a potent antineoplastic efficacy ofLyeTx I-b.

Novel self-nanoemulsifying drug-delivery system enhances antileukemic properties of all-trans retinoic acid.

Aim: All-trans retinoic acid (ATRA) shows erratic oral bioavailability when administered orally against leukemia, which can be solved through its incorporation in self-nanoemulsifying drug-delivery systems (SEDDS). The SEDDS developed contained a hydrophobic ion pair between benzathine (BZT) and ATRA and was enriched with tocotrienols by the input of a palm oil tocotrienol rich fraction (TRF) in its composition. Results: SEDDS-TRF-ATRA-BZT allowed the formation of emulsions with nanometric size that retained ATRA within their core after dispersion. Pharmacokinetic parameters after oral administration of SEDDS-TRF-ATRA-BZT in mice were improved compared with what was seen for an ATRA solution. Moreover, SEDDS-TRF-ATRA-BZT had improved activity against HL-60 cells compared with SEDDS without TRF. Conclusion: SEDDS-TRF-ATRA-BZT is a promising therapeutic choice over ATRA conventional medicine.

A simple ECG marker for the detection of coronary restenosis after successful coronary angioplasty.

BACKGROUND: The dynamic nature of QT dispersion (QTd) and the absence of an acceptable normal value suggest that the relative change not the absolute value of QTd would be a better predicator of coronary restenosis. We sought to examine the usefulness of the relative change in QTd, compared to the absolute value, as a predictor for coronary restenosis after previously successful percutaneous transluminal coronary angioplasty (PTCA). METHODS: Ninety-two patients with a history of successful PTCA who were referred for coronary angiography (CA) for exclusion of coronary restenosis were included in this analysis. QTd was calculated as the difference in milliseconds between the maximum and minimum QT interval in the 12-lead ECG. Relative change in QTd was measured as [QTd at the time of angiography (current) - QTd after the successful PTCA (baseline)]/QTd (baseline) %. Receiver operating characteristics (ROC) analysis was used to detect the best cut-off point and also to compare the diagnostic accuracy of the relative change in QTd vs. the absolute QTd for prediction of coronary restenosis. RESULTS: The relative change in QTd showed a significantly larger ROC area under curve (AUC) compared to the absolute QTd [AUC (95% CI): 0.79 (0.698, 0.872) and 0.61 (0.498, 0.703) respectively; p=0.011 for AUCs comparison]. The best cut-off point for the relative QTd was 40%, and for the absolute QTd this was 50 ms. The sensitivity and specificity of > or =40% increase in QTd to detect coronary restenosis was 71% and 83%, with positive and negative predictive values of 90% and 57%, respectively. The diagnostic accuracy of the absolute value of QTd was much less than this; the sensitivity and specificity of QTd > or =50 ms were 48% and 58%, with positive and negative predictive values of 71% and 34%, respectively. CONCLUSIONS: Compared to the absolute value, the relative change in QTd is a better predictor of coronary restenosis after a previously successful PTCA. These findings may open the door for rethinking the use of QTd as a simple ECG predictor for cardiovascular outcomes.