Design, Synthesis, and Antitumor Evaluation of an Opioid Growth Factor Bioconjugate Targeting Pancreatic Ductal Adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) presents a formidable challenge with high lethality and limited effective drug treatments. Its heightened metastatic potential further complicates the prognosis. Owing to the significant toxicity of current chemotherapeutics, compounds like [Met5]-enkephalin, known as opioid growth factor (OGF), have emerged in oncology clinical trials. OGF, an endogenous peptide interacting with the OGF receptor (OGFr), plays a crucial role in inhibiting cell proliferation across various cancer types. This in vitro study explores the potential anticancer efficacy of a newly synthesized OGF bioconjugate in synergy with the classic chemotherapeutic agent, gemcitabine (OGF-Gem). The study delves into assessing the impact of the OGF-Gem conjugate on cell proliferation inhibition, cell cycle regulation, the induction of cellular senescence, and apoptosis. Furthermore, the antimetastatic potential of the OGF-Gem conjugate was demonstrated through evaluations using blood platelets and AsPC-1 cells with a light aggregometer. In summary, this article demonstrates the cytotoxic impact of the innovative OGF-Gem conjugate on pancreatic cancer cells in both 2D and 3D models. We highlight the potential of both the OGF-Gem conjugate and OGF alone in effectively inhibiting the ex vivo pancreatic tumor cell-induced platelet aggregation (TCIPA) process, a phenomenon not observed with Gem alone. Furthermore, the confirmed hemocompatibility of OGF-Gem with platelets reinforces its promising potential. We anticipate that this conjugation strategy will open avenues for the development of potent anticancer agents.

The pyrazole derivative of usnic acid inhibits the proliferation of pancreatic cancer cells in vitro and in vivo.

BACKGROUND: Pancreatic cancer is one of the leading causes of cancer death in Western societies. Its late diagnosis and resistance to chemotherapies result in a high mortality rate; thus, the development of more effective therapies for the treatment of pancreatic cancer is strongly warranted. Usnic acid (UA) is a secondary metabolite of lichens that shows modest antiproliferative activity toward cancer cells. Recently, we reported the synthesis of a UA pyrazole derivative, named 5, which was more active than the parent compound toward cervical cancer cells. Here, its anticancer potential has been evaluated in detail in other cancer cells, particularly pancreatic cancer cells. METHODS: The impact of UA and derivative 5 on cell viability, morphology, cell cycle, and death was assessed using the MTT test, electron microscopy, flow cytometry, and immunoblotting, respectively. The calcium ions level was detected fluorometrically. In vivo, the anticancer activity of 5 was evaluated in a murine xenograft model. RESULTS: Derivative 5 inhibited the viability of different cancer cells. Noncancerous cells were less sensitive. It induced the release of calcium ions from the endoplasmic reticulum (ER) and ER stress, which was manifested by cell vacuolization. It was accompanied by G0/G1 cell cycle arrest and cell death of pancreatic cancer cells. When applied to nude mice with xenografted pancreatic cancer cells, 5 inhibited tumor growth, with no signs of kidney or liver toxicity. CONCLUSIONS: UA derivative 5 is superior to UA inhibiting the growth and proliferation of pancreatic cancer cells. ER stress exaggeration is a mechanism underlying the activity of derivative 5.

Estimation of smokers' exposure to mercury from combustible tobacco products, based on the approach used in food consumers' exposure estimation.

Smoking has been known to mankind for centuries, but it is only in recent decades that much attention has been paid to the harmfulness of this habit. Mercury inhalation is particularly dangerous in this respect and smoking creates extremely favorable conditions for the emission and targeted delivery of this element into the lungs. Despite this fact, a lack of a clear method for estimating the exposure of tobacco consumers to mercury was identified. This work shows justification to transfer the approach of estimating food product consumers' exposure to estimate the exposure of combustible tobacco product consumers to this element. In addition, it was noted that researchers' attention is mainly focused on cigarettes, while the tobacco market has a wide range of combustible products. Therefore, in this work, the mercury content of cigars (8.45 ± 0.18-41.02 ± 0.20 µg/kg), pipe tobaccos (8.03 ± 0.52-25.48 ± 0.50 µg/kg), bidis (14.93 ± 0.47-31.79 ± 0.26 µg/kg) and cigarette tobaccos (14.22 ± 0.71-34.5 ± 1.4 µg/kg) was analyzed. This study demonstrates that smoking can contribute significant total mercury exposure to consumers', although it is unlikely to cause mercury poisoning regardless of other exposure sources.

The Use of a Barley-Based Well to Define Cationic Betaglucan to Study Mammalian Cell Toxicity Associated with Interactions with Biological Structures.

Among potential macromolecule-based pharmaceuticals, polycations seem particularly interesting due to their proven antimicrobial properties and use as vectors in gene therapy. This makes an understanding of the mechanisms of these molecules' interaction with living structures important, so the goal of this paper was to propose and carry out experiments that will allow us to characterize these phenomena. Of particular importance is the question of toxicity of such structures to mammalian cells and, in the work presented here, two lines, normal fibroblasts 3T3-L1 and A549 lung cancer, were used to determine this. In this work, three well-defined cationic derivatives of barley-derived betaglucans obtained in a reaction with glycidyltrimethylammonium chloride (BBGGTMAC) with different degrees of cationization (50, 70, and 100% per one glucose unit) and electrostatic charge were studied. The studies address interactions of these polymers with proteins (bovine serum proteins and BSA), nucleic acids (DNA), glycosaminoglycans (heparin), and biological membranes. The results described in this study make it possible to indicate that toxicity is most strongly influenced by interactions with biological membranes and is closely related to the electrostatic charge of the macromolecule. The presentation of this observation was the goal of this publication. This paper also shows, using fluorescently labeled variants of polymers, the penetration and impact on cell structure (only for the polymer with the highest substitution binding to cell membranes is observed) by using confocal and SEM (for the polymer with the highest degree of substitution, and the appearance of additional structures on the surface of the cell membrane is observed). The labeled polymers are also tools used together with dynamic light scattering and calorimetric titration to study their interaction with other biopolymers. As for the interactions with biological membranes, lipid Langmuir monolayers as model membrane systems were used.

Centripetal Nystagmus, Slow Saccades, Cerebellar Ataxia, and Parkinsonism in a Patient With Anti-GAD65-Associated Stiff Person Syndrome Spectrum Disorder.

ABSTRACT: A 68-year-old woman with positional dizziness and progressive imbalance presented for vestibular evaluation. Examination was notable for spontaneous downbeat nystagmus (DBN), horizontal and vertical gaze-evoked nystagmus (GEN) with centripetal and rebound nystagmus, and positional apogeotropic nystagmus. There was also mild-moderate slowing of saccades horizontally and vertically and poor fast phases with an optokinetic stimulus. Further consultation by a movement disorder specialist uncovered asymmetric decrementing bradykinesia and rigidity, masked facies, and a wide-based stance without camptocormia. Screening serum laboratory results for metabolic, rheumatologic, infectious, heavy metal, endocrine, or vitamin abnormalities was normal. Surveillance imaging for neoplasms was unremarkable, and cerebrospinal fluid (CSF) analysis was negative for 14-3-3 and real-time quaking-induced conversion (RT-QuIC). However, her anti-glutamic acid decarboxylase-65 (GAD65) immunoglobulin G (IgG) level was markedly elevated in serum to 426,202 IU/mL (reference range 0-5 IU/mL) and in CSF to 18.1 nmol/L (reference range <0.03 nmol/L). No other autoantibodies were identified on the expanded paraneoplastic panel. The patient was referred to neuroimmunology, where torso rigidity, spasticity, and significant paravertebral muscle spasms were noted. Overall, the clinical presentation, examination findings, and extensive workup were consistent with a diagnosis of anti-GAD65-associated stiff person syndrome-plus (musculoskeletal plus cerebellar and/or brainstem involvement). She was subsequently treated with intravenous immunoglobulin (IVIg) and has been stable since commencing this therapy. In patients with centripetal nystagmus, especially in association with other cerebellar findings, an autoimmune cerebellar workup should be considered.

Frequency And Factors Associated With Adverse Reactions After Administration of Inactivated COVID-19 Vaccine Among Health Workers

ABSTRACT Background: The 2019 coronavirus (COVID-19) has precipitated a significant public health crisis. Our study aimed to evaluate the prevalence and risk factors associated with adverse reactions to the inactivated CoronaVac vaccine. Methods: The study involved voluntary health workers who received CoronaVac vaccine. We documented the sociodemographic information of 2,019 participants who volunteered for our study. Of these, 1,964 and 1,702 participants were interviewed by phone 1 month after the first and second dose, respectively, during which they were queried about any adverse reactions. Results: Within the first week after the first dose, adverse reactions were observed in 856 (43.3%) participants, with 133 (6.7%) experiencing them during the second week, and 96 (4.9%) people at the end of the first month. For the second dose, 276 individuals (16.2%) reported adverse reactions. The prevalence of both local and systemic adverse events ranged from 9.5-11.2% overall. Fatigue was the most common adverse reaction overall, while pain at the injection site was the most frequent local adverse reaction. Conclusions: The evaluation of both systemic and local side effects revealed no significant adverse reactions to the inactivated CoronaVac vaccine (Sinovac Life Sciences, Beijing, China). Our study found that the incidence of systemic and local adverse responses to the CoronaVac vaccination was lower than the rates reported in studies involving the recombinant adenovirus type-5, BNT162b1, and ChAdOx1nCoV-19 COVID-19 vaccines, all of which underwent the World Health Organization LULUC/PQ evaluation process.

Effects of perilipin-5 on lipid metabolism and high-sensitivity cardiac troponin I

SUMMARY OBJECTIVE: Heart attack is one of the most common causes of sudden death in adults. Therefore, early detection of heart attack and investigation of potential new biomarkers are of great importance. We investigated whether perilipin-5 is a potential biomarker by examining changes in perilipin-5 serum levels along with high-sensitivity cardiac troponin I during a heart attack. METHODS: The subjects were divided into two groups: (1) control group and (2) patients with heart attack, with 150 people in each group. High-sensitivity cardiac troponin I, perilipin-5, total oxidant status, malondialdehyde, reduced glutathione, and superoxide dismutase levels in serum samples were measured. In addition, perilipin-5 mRNA expressions and protein levels were analyzed. RESULTS: There was no overall statistical difference between the demographic characteristics of the groups. However, high-density lipoprotein, creatine kinase, Creatine kinase myocardial band, aspartate amino transferase, lactate dehydrogenase, and calcium levels were higher in the heart attack group compared to the control group. We found that the high-sensitivity cardiac troponin I and perilipin-5 levels increased in the patients with heart attack (p<0.0001) compared to control. Although there was an insignificant increase in malondialdehyde levels in the heart attack group (p>0.05), there was a 35.9% increase in total oxidant status levels and a 33.5 and 24.1% decrease in glutathione and superoxide dismutase levels, respectively (p<0.01), compared to control. Perilipin-5 mRNA and protein levels in heart attack patients increased by 48.2 and 23.6%, respectively, compared to the control group (p<0.01). CONCLUSION: Our results showed that perilipin-5 together with high-sensitivity cardiac troponin I could be a promising biomarker in heart attack.

Olfactory rehabilitation and olfactory bulb volume changes in patients after total laryngectomy: a prospective randomized study / Reabilitação olfatória e alterações do volume do bulbo olfatório em pacientes pós-laringectomia total: um estudo prospectivo randomizado

Abstract Introduction: After total laryngectomy, decreased olfactory function and olfactory bulb volume shrinkage have been reported to occur due to olfactory deprivation caused by nasal airflow interruption. There is evidence that the olfactory system can be modulated by repeated exposure to odors in a procedure called olfactory training. However, it is not known whether any recovery of the lost olfactory bulb volume is possible by eliminating olfactory deprivation via olfactory rehabilitation long after laryngectomy. Objective: This study examined the recovery of olfactory function and the change in olfactory bulb volume via long-term olfactory rehabilitation after total laryngectomy. Methods: Possible causes of olfactory dysfunction in the study participants were evaluated by collecting detailed anamnesis. As olfactory tests, orthonasal butanol threshold and odor discrimination tests were performed. Three-dimensional olfactory bulb volumes were calculated using manual segmentation on T2-weighted coronal magnetic resonance images. In olfactory rehabilitation, four different odors were applied to all patients orthonasally, using a larynx bypass technique for 30 min per day for 6 months. Olfactory tests were performed before the rehabilitation and after 6 months of rehabilitation, and olfactory bulb volume measurements were performed using magnetic resonance images. Results: Eleven patients diagnosed with advanced laryngeal cancer who underwent total laryngectomy and postoperative radiotherapy with a follow-up of 5-10 years were included in the study. All patients were male, and the mean age was 58.18 ±4.17 years. In total laryngectomized patients, the olfactory bulb volumes measured on magnetic resonance images were 42.25 ± 12.8 mm3 before and 55.5 ± 11.22 mm3 after rehabilitation, and this increase was highly significant. Olfactory test scores were 2.3 ± 1.27 before and 4.39 ± 0.86 after rehabilitation, and this increase was also highly significant. Conclusion: As a result of the olfactory rehabilitation applied by providing orthonasal air flow, the olfactory function lost after total laryngectomy was improved considerably, and the olfactory bulb volume was significantly increased. The increase in olfactory bulb volume in total laryngectomy patients via olfactory rehabilitation to eliminate olfactory deprivation due to nasal airflow interruption was demonstrated for the first time in this prospective longitudinal study.

Resumo Introdução: Após a laringectomia total, foi relatada a ocorrência de diminuição da função olfatória e redução do volume do bulbo olfatório devido à privação olfatória causada pela interrupção do fluxo aéreo nasal. Há evidências de que o sistema olfatório pode ser modulado pela exposição repetida a odores em um procedimento denominado treinamento olfatório. Entretanto, não se sabe se qualquer grau de recuperação do volume perdido do bulbo olfatório é possível ao eliminar a privação olfatória através de reabilitação muito tempo depois da laringectomia. Objetivo: Este estudo avaliou a recuperação da função olfatória e a mudança no volume do bulbo olfatório através da reabilitação olfatória de longo prazo após a laringectomia total. Métodos: As possíveis causas de disfunção olfatória nos participantes do estudo foram avaliadas através da anamnese detalhada. Como testes olfatórios, foram feitos os testes de limiar de butanol ortonasal e de discriminação de odores. Os volumes tridimensionais do bulbo olfatório foram calculados com segmentação manual em imagens de ressonância magnética coronal ponderadas em T2. Na reabilitação olfatória, quatro odores diferentes foram aplicados a todos os pacientes ortonasalmente com uma técnica de bypass laríngeo por 30 minutos por dia durante 6 meses. Os testes olfatórios foram feitos antes da reabilitação e 6 meses após a reabilitação e as medidas do volume do bulbo olfatório foram feitas por imagens de ressonância magnética. Resultados: Foram incluídos no estudo 11 pacientes com diagnóstico de câncer de laringe avançado, submetidos à laringectomia total e radioterapia pós-operatória em um seguimento de 5 a 10 anos. Todos os pacientes eram do sexo masculino e a média de idade foi de 58,18 ± 4,17 anos. Em pacientes com laringectomia total, os volumes do bulbo olfatório medidos por imagens de ressonância magnética foram de 42,25 ± 12,8 mm3 antes e 55,5 ± 11,22 mm3 após a reabilitação e esse aumento foi altamente significante. Os escores dos testes olfatórios foram 2,3 ± 1,27 antes e 4,39 ± 0,86 após a reabilitação e esse aumento também foi altamente significante. Conclusão: Como resultado da reabilitação olfatória aplicada através do fornecimento de fluxo de ar ortonasal, a função olfatória perdida após a laringectomia total melhorou consideravelmente e o volume bulbo olfatório mostrou aumento significativo. O aumento no volume do bulbo olfatório em pacientes submetidos a laringectomia total através da reabilitação olfatória para eliminar a privação olfatória devido à interrupção do fluxo aéreo nasal foi demonstrado pela primeira vez neste estudo longitudinal prospectivo.

Tumor Budding Is an Independent Prognostic Factor in Pancreatic Adenocarcinoma and It Positively Correlates with PD-L1 Expression on Tumor Cells.

Pancreatic adenocarcinoma is one of the leading causes of cancer-related death in developed countries. Only 15% of patients are candidates for radical surgery, and adequate prognostication may guide proper postsurgical management. We aimed to retrospectively assess the prognostic significance of the immunohistochemical expression of immune checkpoint receptors (PD-L1 and VISTA), markers of systemic inflammation, thrombosis in the tumor area, and the tumor budding in the group of 107 patients diagnosed with pancreatic adenocarcinoma in a single center. The high expression of PD-L1 on tumor cells (TCs) was associated with worse overall survival (OS, p = 0.041, log-rank). On the contrary, high PD-L1 or VISTA on tumor-associated immune cells (TAICs) was correlated with better OS (p = 0.006 and p = 0.008, respectively, log-rank). The joint status of PD-L1 on TCs and TAICs stratified patients into three prognostic groups. The cases with high-grade budding were characterized by higher PD-L1 expression on TCs (p = 0.008) and elevated systemic inflammatory markers. Moreover, budding was identified as the independent prognostic factor in multivariate Cox regression analysis (HR = 2.87; 95% CI = 1.75−4.68; p < 0.001). To conclude, the pattern of PD-L1 and VISTA expression was associated with survival in univariate analysis. Tumor budding accurately predicts outcomes in pancreatic cancer and should be incorporated into routine histopathological practice.

Review of cigars and cigar-type products as potential sources of consumer exposure to heavy metals.

The popularity of cigars, growing since 1993, has not gone hand in hand with the increased interest of researchers in these products. Although the literature widely describes the harmfulness of tobacco and the content of toxic substances in tobacco products, the topic is often treated selectively as relating primarily to cigarettes and rarely extends to other products of the broadly defined tobacco industry. However, there is no reason to marginalize the harmful effects of other nicotine products, (which include tobacco products such as cigars). The study analyzed the available literature on the content of selected heavy metals in cigar tobacco. Among the heavy metals, the following contents of elements in tobacco were recorded in cigars: Fe (420-2200 µg/g), Mn (100-370 µg/g), Zn (14-180 µg/g), Cu (15-140 µg/g), Pb (not detected-32 µg/g), Cd (nd-19 µg/g), Ni (nd-13 µg/g), Cr (nd-10 µg/g), Co (0.65-1.0 µg/g), As (nd-0.66 µg/g), Hg (18-25 ng/g). Importantly, the values often differ between cigars of different origins and types, indicating the need for more extensive research.

Kidney Transplantation in Patients After Bariatric Surgery: High-Volume Bariatric and Transplant Center Experience.

BACKGROUND: Kidney transplantation (KTx) is the best type of treatment for patients with end-stage renal disease (ESRD). Unfortunately, obesity may be a contraindication for transplantation. Our study aimed to evaluate the results of KTx in patients who had bariatric surgery (BS) prior to transplantation. METHODS: A single center, with experience in bariatric and transplant surgery, presents a retrospective study of 13 patients who received a kidney transplant after a gastric bypass (GB) operation between 2012 and 2019. RESULTS: Thirteen patients, who were potential candidates for KTx, were previously qualified for BS because of a body mass index (BMI) > 35 kg/m2. Additionally, all patients had arterial hypertension, 60% of patients had diabetes, and 30% of patients had coronary artery disease. Patients were activated on the waiting list when their BMI was < 35 kg/m2. KTx was performed between 5 and 29 months after BS. One patient needed reoperation due to a urinary leak and another patient needed reoperation because of a high-pressure lymphocele. We diagnosed 2 delayed graft functions (DGFs) and 1 acute rejection. One patient died for reasons independent of surgery. The KTx observation period ranged from 3 to 8 years. Currently, 11 patients has stable renal function: creatinine concentration is 0.8-1.8 mg/dL and BMI is between 23 and 35 kg/m2. CONCLUSIONS: Despite the small group of patients, we can assume that kidney transplantation can be safely performed in patients with end-stage renal disease (ESRD) who have previously undergone gastric bypass (GB) as a graft bridging procedure. In some cases, BS may be the only chance of getting an organ.

Comprehensive cancer-oriented biobanking resource of human samples for studies of post-zygotic genetic variation involved in cancer predisposition.

The progress in translational cancer research relies on access to well-characterized samples from a representative number of patients and controls. The rationale behind our biobanking are explorations of post-zygotic pathogenic gene variants, especially in non-tumoral tissue, which might predispose to cancers. The targeted diagnoses are carcinomas of the breast (via mastectomy or breast conserving surgery), colon and rectum, prostate, and urinary bladder (via cystectomy or transurethral resection), exocrine pancreatic carcinoma as well as metastases of colorectal cancer to the liver. The choice was based on the high incidence of these cancers and/or frequent fatal outcome. We also collect age-matched normal controls. Our still ongoing collection originates from five clinical centers and after nearly 2-year cooperation reached 1711 patients and controls, yielding a total of 23226 independent samples, with an average of 74 donors and 1010 samples collected per month. The predominant diagnosis is breast carcinoma, with 933 donors, followed by colorectal carcinoma (383 donors), prostate carcinoma (221 donors), bladder carcinoma (81 donors), exocrine pancreatic carcinoma (15 donors) and metachronous colorectal cancer metastases to liver (14 donors). Forty percent of the total sample count originates from macroscopically healthy cancer-neighboring tissue, while contribution from tumors is 12%, which adds to the uniqueness of our collection for cancer predisposition studies. Moreover, we developed two program packages, enabling registration of patients, clinical data and samples at the participating hospitals as well as the central system of sample/data management at coordinating center. The approach used by us may serve as a model for dispersed biobanking from multiple satellite hospitals. Our biobanking resource ought to stimulate research into genetic mechanisms underlying the development of common cancers. It will allow all available "-omics" approaches on DNA-, RNA-, protein- and tissue levels to be applied. The collected samples can be made available to other research groups.

Influence of Hypoxia on Radiosensitization of Cancer Cells by 5-Bromo-2'-deoxyuridine.

Radiotherapy is a crucial cancer treatment, but its outcome is still far from satisfactory. One of the reasons that cancer cells show resistance to ionizing radiation is hypoxia, defined as a low level of oxygenation, which is typical for solid tumors. In the hypoxic environment, cancer cells are 2-3 times more resistant to ionizing radiation than normoxic cells. To overcome this important impediment, radiosensitizers should be introduced to cancer therapy. When modified with an electrophilic substituent, nucleosides may undergo efficient dissociative electron attachment (DEA) that leaves behind nucleoside radicals, which, in secondary reactions, are able to induce DNA damage, leading to cancer cell death. We report the radiosensitizing effect of one of the best-known DEA-type radiosensitizers-5-bromo-2'-deoxyuridine (BrdU)-on breast (MCF-7) and prostate (PC3) cancer cells under both normoxia and hypoxia. MCF-7 and PC3 cells were treated with BrdU to investigate the effect of hypoxia on cell proliferation, incorporation into DNA and radiosensitivity. While the oxygen concentration did not significantly affect the efficiency of BrdU incorporation into DNA or the proliferation of tumor cells, the radiosensitizing effect of BrdU on hypoxic cells was more evident than on normoxic cells. Further mechanistic studies performed with the use of flow cytometry showed that under hypoxia, BrdU increased the level of histone H2A.X phosphorylation after X-ray exposure to a greater extent than under normal oxygenation conditions. These results confirm that the formation of double-strand breaks in hypoxic BrdU-treated cancer cells is more efficient. In addition, by performing stationary radiolysis of BrdU solution in the presence of an ●OH radical scavenger, we compared the degree of its electron-induced degradation under aerobic and anaerobic conditions. It was determined that radiodegradation under anaerobic conditions was almost twice as high as that under aerobic conditions.

Benefitial role of electrochemotherapy in locally advanced pancreatic cancer - radiological perspective.

Irreversible electroporation (IRE) of locally advanced pancreatic cancer is an increasingly used method for unresectable pancreatic cancer that can be used in cytoreduction followed by surgical treatment and shows promising results in palliative care. IRE is an ablative technique where electric pulses cause damage to the cell membrane leading to apoptosis without the destruction of stroma. The application of IRE increases the concentration of hydrophobic regimens like bleomycin within the tumor, what could improve the effectiveness of treatment. This fusion of those two treatments is called electrochemotherapy. In this review, the authors will discuss the radiological perspective of possible beneficial role of irreversible electroporation in relation with chemotherapy in pancreatic cancer treatment.

The Isoxazole Derivative of Usnic Acid Induces an ER Stress Response in Breast Cancer Cells That Leads to Paraptosis-like Cell Death.

Derivatives of usnic acid (UA), a secondary metabolite from lichens, were synthesized to improve its anticancer activity and selectivity. Recently we reported the synthesis and activity of an UA isoxazole derivative, named 2b, against cancer cells of different origins. Herein, the molecular mechanisms underlying its activity and efficacy in vivo were tested. The viability of breast cancer or normal cells has been tested using an MTT assay. Cell and organelle morphology was analyzed using light, electron and fluorescence microscopy. Gene expression was evaluated by RNAseq and protein levels were evaluated by Western blotting. In vivo anticancer activity was evaluated in a mice xenograft model. We found that 2b induced massive vacuolization which originated from the endoplasmic reticulum (ER). ER stress markers were upregulated both at the mRNA and protein levels. ER stress was caused by the release of Ca2+ ions from the ER by IP3R channels which was mediated, at least partly, by phospholipase C (PLC)-synthetized 1,4,5-inositol triphosphate (IP3). ER stress led to cell death with features of apoptosis and paraptosis. When applied to nude mice with xenografted breast cancer cells, 2b stopped tumour growth. In mice treated with 2b, vacuolization was observed in tumour cells, but not in other organs. This study shows that the antiproliferative activity of 2b relates to the induction of ER stress in cancer, not in healthy, cells and it leads to breast cancer cell death in vitro and in vivo.

Clinical Problem-Solving: Lower Extremity Weakness & Paresthesia in an Immunocompromised Patient With a Complex Cancer History.

We present a case of new onset bilateral lower extremity weakness, paresthesia, urinary retention and bowel incontinence in a 51-year-old man. He had a complicated history of acute myelogenous leukemia with known central nervous system (CNS) and leptomeningeal involvement status post allogenic stem cell transplant complicated by chronic graft versus host disease (GVHD). We review the differential diagnosis as the physical exam and diagnostic results evolved. We also provide a review of the relevant literature supporting our favored diagnosis, as well as other competing diagnoses in this complicated case. The ultimate differential diagnosis included viral myelitis, treatment-related myelopathies, and CNS GVHD. The case provides a sobering reminder that even with an appropriate diagnostic workup, some cases remain refractory to therapeutic efforts. It also underscores the importance of a sensitive neurologic exam, given the significant clinico-radiological delay, and reviews the complex differential diagnosis for myelopathy.

Erythropoietin Concentration in Boys With p.His63Asp Polymorphism of the HFE Gene.

The molecular mechanism that regulates iron homeostasis is based on a network of signals, which reflect on the iron requirements of the body. HFE-related hemochromatosis is characterized by excessive intestinal absorption of dietary iron, in particular cases resulting in pathologically high iron storage in tissues and organs. During childhood, HFE gene homozygosity or heterozygosity manifests exclusively in the form of biochemical abnormalities. Because of their mutual link, bioavailable iron and endogenous erythropoietin (EPO) are indispensable for effective erythropoiesis. We analyzed the impact of p.(His63Asp) polymorphism of the HFE gene on erythropoiesis taking into consideration endogenous EPO production in the developmental age. In the study we performed, we observed a significant, strong and negative correlation between the concentration of EPO, hemoglobin, and red blood cell count. A negative trend was also noted on the impact of iron concentration and transferrin saturation on EPO production. In conclusion, this preliminary study demonstrates an impaired impact of endogenous EPO on erythropoiesis in the presence of increased iron content in carriers of p.(His63Asp) (heterozygotes) variant of the HFE gene in developmental age.

Synthesis and in Vitro Toxicity Assessment of Different Nano-Calcium Phosphate Nanoparticles

Abstract Nanoscale biomaterials are commonly used in a wide range of biomedical applications such as bone graft substitutes, gene delivery systems, and biologically active agents. On the other hand, the cytotoxic potential of these particles hasn't yet been studied comprehensively to understand whether or not they exert any negative impact on the cellular structures. Here, we undertook the synthesis of beta-tricalcium phosphate (ß-TCP) and biphasic tricalcium phosphate (BCP) nanoparticles (NPs) and determine their concentration-dependent toxic effects in human fetal osteoblastic (hFOB 1.19) cell line. Firstly, BCP and β-TCP were synthesized using a water-based precipitation technique and characterized by X-Ray Diffraction (XRD), Raman Spectroscopy, and Transmission Electron Microscopy (TEM). The cytological effects of β-TCP and BCP at different concentrations (0-640 ppm) were evaluated by using 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays. The total oxidative status (TOS) parameter was used for investigating oxidative stress potentials of the NPs. In addition, the study assessed the DNA damage product 8-hydroxy-2′-deoxyguanosine (8-Oxo-dG) level in hFOB 1.19 cell cultures. The results indicated that the β-TCP (above 320 ppm) and BCP (above 80 ppm) NPs exhibited cytotoxicity effects on high concentrations. It was also observed that the oxidative stress increased relatively as the concentrations of NPs increased, aligning with the cytotoxicity results. However, the NPs concentrations of 160 ppm and above increased the level of 8-OH-dG. Consequently, there is a need for more systematic in vivo and in vitro approaches to the toxic effects of both nanoparticles.

Scalp cooling for reducing alopecia in gynecology oncology patients treated with dose-dense chemotherapy: A pilot project.

OBJECTIVE: Determine the efficacy of scalp cooling for the prevention of chemotherapy-induced alopecia in gynecology oncology patients. METHODS: This prospective pilot study included patients diagnosed with a gynecological malignancy that received DigniCap™ scalp cooling. Patients were divided into two groups based on chemotherapy regimen: Carboplatin with area under the curve (AUC) 5-6 every three weeks and (1) conventional Paclitaxel 175 mg/m2 every three weeks or (2) Paclitaxel 80 mg/m2 weekly. A 1-10 visual analogue scale (1 no hair loss, 10 - complete hair loss) was used to assess degree of hair loss by patients themselves and by a certified dermatologist using photographs. Changes in quality of life and body image were measured using the European Organization for Research and Treatment of Cancer quality of life questionnaire version 3 (EORTC QLQ-C30) and the Body Image Scale (BIS) for cancer patients. RESULTS: Hair preservation occurred with use of a scalp cooling device for patients receiving weekly Paclitaxel (n = 20), but not conventional every three weeks Paclitaxel (n = 8). Ten of 15 patients (66.7%) in the dose-dense group lost less than 50% of their hair based on self-assessment and 14 of 16 (87.5%) based on dermatologist assessment. No patient in this group acquired a cranial prosthesis (wig). There was no difference between groups in terms of quality of life (QoL) and BIS scores. CONCLUSION: Scalp cooling may allow for hair preservation in gynecology oncology patients receiving Carboplatin AUC 5-6 and weekly Paclitaxel 80 mg/m2 combination chemotherapy.