Dual-targeting exosomes for improved drug delivery in breast cancer.

Aims: The authors investigated whether displaying more than one homing peptide enhanced the tumor-targeting efficiency of exosomes. Materials & methods: Exosomes from human embryonic kidney cells (HEK293F) were engineered to display either mono- or dual-tumor-penetrating peptides, iRGD and tLyp1. Exosomes were purified via tangential flow filtration followed by ultracentrifugation. Results: When loaded with doxorubicin (Dox), the dual iRGD-tLyp1 exosomes strongly enhanced Dox uptake in both MCF-7 and MDA-MB-231 breast cancer cell lines, superior to single iRGD or tLyp1 exosomes. The dual iRGD-tLyp1 exosomal Dox was also the most potent, with IC50/GI50 values being 3.7-17.0-times lower than those of free Dox and other exosomal Dox. Conclusion: Selecting appropriate combinatorial homing peptides could be an approach for future precision nanomedicine.

Cystathionine-ß-synthase X proteins negatively regulate NADPH-thioredoxin reductase C activity.

Redox regulation is a posttranslational modification based on the redox reaction of protein thiols. A small ubiquitous protein thioredoxin (Trx) plays a central role in redox regulation, but a unique redox-regulatory factor called NADPH-Trx reductase C (NTRC) is also found in plant chloroplasts and some cyanobacteria. Several important functions of NTRC have been suggested, but the mechanism for controlling NTRC activity remains undetermined. Cystathionine-ß-synthase X (CBSX) proteins have been previously shown to interact with NTRC physically. Based on these observations, this study biochemically investigated the functional interaction between CBSX proteins and NTRC from Arabidopsis thaliana in vitro. Consequently, we concluded that CBSX proteins act as negative regulators of NTRC in the presence of AMP.

Survey of Protein Sequence Embedding Models.

Derived from the natural language processing (NLP) algorithms, protein language models enable the encoding of protein sequences, which are widely diverse in length and amino acid composition, in fixed-size numerical vectors (embeddings). We surveyed representative embedding models such as Esm, Esm1b, ProtT5, and SeqVec, along with their derivatives (GoPredSim and PLAST), to conduct the following tasks in computational biology: embedding the Saccharomyces cerevisiae proteome, gene ontology (GO) annotation of the uncharacterized proteins of this organism, relating variants of human proteins to disease status, correlating mutants of beta-lactamase TEM-1 from Escherichia coli with experimentally measured antimicrobial resistance, and analyzing diverse fungal mating factors. We discuss the advances and shortcomings, differences, and concordance of the models. Of note, all of the models revealed that the uncharacterized proteins in yeast tend to be less than 200 amino acids long, contain fewer aspartates and glutamates, and are enriched for cysteine. Less than half of these proteins can be annotated with GO terms with high confidence. The distribution of the cosine similarity scores of benign and pathogenic mutations to the reference human proteins shows a statistically significant difference. The differences in embeddings of the reference TEM-1 and mutants have low to no correlation with minimal inhibitory concentrations (MIC).

IL-33 promotes gastric tumour growth in concert with activation and recruitment of inflammatory myeloid cells.

Interleukin-33 (IL-33) is an IL-1 family cytokine known to promote T-helper (Th) type 2 immune responses that are often deregulated in gastric cancer (GC). IL-33 is overexpressed in human gastric tumours suggesting a role in driving GC progression although a causal link has not been proven. Here, we investigated the impact of IL-33 genetic deficiency in the well-characterized gp130 F/F mouse model of GC. Expression of IL-33 (and it's cognate receptor, ST2) was increased in human and mouse GC progression. IL-33 deficient gp130 F/F /Il33 -/- mice had reduced gastric tumour growth and reduced recruitment of pro-tumorigenic myeloid cells including key mast cell subsets and type-2 (M2) macrophages. Cell sorting of gastric tumours revealed that IL-33 chiefly localized to gastric (tumour) epithelial cells and was absent from tumour-infiltrating immune cells (except modest IL-33 enrichment within CD11b+ CX3CR1+CD64+MHCII+ macrophages). By contrast, ST2 was absent from gastric epithelial cells and localized exclusively within the (non-macrophage) immune cell fraction together with mast cell markers, Mcpt1 and Mcpt2. Collectively, we show that IL-33 is required for gastric tumour growth and provide evidence of a likely mechanism by which gastric epithelial-derived IL-33 drives mobilization of tumour-promoting inflammatory myeloid cells.

A Curious Case of Hypercalcemia.

Humoral hypercalcemia of malignancy is rarely seen in urothelial cancer. In this report, we present a case of an 81-year-old female patient who presented with a markedly elevated calcium level leading to severe altered mental status and was found to have urothelial cancer. To our knowledge, only three cases of urothelial carcinoma with squamous cell differentiation have been reported.

Polypyrrole with Phosphor Tungsten Acid and Carbide-Derived Carbon: Change of Solvent in Electropolymerization and Linear Actuation.

Linear actuators based on polypyrrole (PPy) are envisaged to have only one ion that triggers the actuation direction, either at oxidation (anion-driven) or at reduction (cation-driven). PPy doped with dodecylbenzenesulfonate (PPy/DBS) is the most common applied conducting polymer having cation-driven actuation in aqueous solvent and mainly anion-driven actuation in an organic electrolyte. It is somehow desired to have an actuator that is independent of the applied solvent in the same actuation direction. In this research we made PPy/DBS with the addition of phosphorus tungsten acid, forming PPyPT films, as well with included carbide derived carbon (CDC) resulting in PPyCDC films. The solvent in electropolymerization was changed from an aqueous ethylene glycol mixture to pure EG forming PPyPT-EG and PPyCDC-EG composites. Our goal in this study was to investigate the linear actuation properties of PPy composites applying sodium perchlorate in aqueous (NaClO4-aq) and propylene carbonate (NaClO4-PC) electrolytes. Cyclic voltammetry and square potential steps in combination with electro-chemo-mechanical-deformation (ECMD) measurements of PPy composite films were performed. The PPyPT and PPyCDC had mixed ion-actuation in NaClO4-PC while in NaClO4-aq expansion at reduction (cation-driven) was observed. Those novel PPy composites electropolymerized in EG solvent showed independently which solvent applied mainly expansion at reduction (cation-driven actuator). Chronopotentiometric measurements were performed on all composites, revealing excellent specific capacitance up to 190 F g-1 for PPyCDC-EG (best capacitance retention of 90 % after 1000 cycles) and 130 F g-1 for PPyPT-EG in aqueous electrolyte. The films were characterized by scanning electron microscopy (SEM), Raman, Fourier-transform infrared (FTIR) and energy dispersive X-ray spectroscopy (EDX).

Thermal effects in molecular gas-filled hollow-core fibers.

Few-cycle sources with high average powers are required for applications to attosecond science. Raman-enhanced spectral broadening of Yb-doped laser amplifiers in molecular gases can yield few-cycle pulses, but thermal excitation of vibrational and rotational degrees of freedom may preclude high-power operation. Here we investigate changes in the spectral broadening associated with repetitive laser interactions in an ${{\rm{N}}_2}{\rm{O}}$-filled hollow-core fiber. By comparing experimental measurements of the spectrum associated with each laser pulse to simulations based on a density matrix model, we find that losses in a spectral bandwidth and transmission are largely dominated by thermal excitation of the gas.

Pre-conditioning modifies the TME to enhance solid tumor CAR T cell efficacy and endogenous protective immunity.

Chimeric antigen receptor (CAR) T cell therapy has led to impressive clinical responses in patients with hematological malignancies; however, its effectiveness in patients with solid tumors has been limited. While CAR T cells for the treatment of advanced prostate and pancreas cancer, including those targeting prostate stem cell antigen (PSCA), are being clinically evaluated and are anticipated to show bioactivity, their safety and the impact of the immunosuppressive tumor microenvironment (TME) have not been faithfully explored preclinically. Using a novel human PSCA knockin (hPSCA-KI) immunocompetent mouse model, we evaluated the safety and therapeutic efficacy of PSCA-CAR T cells. We demonstrated that cyclophosphamide (Cy) pre-conditioning significantly modified the immunosuppressive TME and was required to uncover the efficacy of PSCA-CAR T cells in metastatic prostate and pancreas cancer models, with no observed toxicities in normal tissues with endogenous expression of PSCA. This combination dampened the immunosuppressive TME, generated pro-inflammatory myeloid and T cell signatures in tumors, and enhanced the recruitment of antigen-presenting cells, as well as endogenous and adoptively transferred T cells, resulting in long-term anti-tumor immunity.

Coordinate expression loss of GKN1 and GKN2 in gastric cancer via impairment of a glucocorticoid-responsive enhancer.

Gastrokines (GKNs) are anti-inflammatory proteins secreted by gastric epithelial (surface mucous and pit) cells, with their aberrant loss of expression causally linked to premalignant inflammation and gastric cancer (GC). Transcriptional mechanisms accounting for GKN expression loss have not been elucidated. Using human clinical cohorts, mouse transgenics, bioinformatics, and transfection/reporter assays, we report a novel mechanism of GKN gene transcriptional regulation and its impairment in GC. GKN1/GKN2 loss is highly coordinated, with both genes showing parallel downregulation during human and mouse GC development, suggesting joint transcriptional control. In BAC transgenic studies, we defined a 152-kb genomic region surrounding the human GKN1/GKN2 genes sufficient to direct their tissue- and lineage-restricted expression. A screen of the 152-kb region for candidate regulatory elements identified a DNase I hypersensitive site (CR2) located 4 kb upstream of the GKN1 gene. CR2 showed overlapping enrichment of enhancer-related histone marks (H3K27Ac), a consensus binding site (GRE) for the glucocorticoid receptor (GR), strong GR occupancy in ChIP-seq data sets and, critically, exhibited dexamethasone-sensitive enhancer activity in reporter assays. Strikingly, GR showed progressive expression loss, paralleling that of GKN1/2, in human and mouse GC, suggesting desensitized glucocorticoid signaling as a mechanism underlying GKN loss. Finally, mouse adrenalectomy studies revealed a critical role for endogenous glucocorticoids in sustaining correct expression (and anti-inflammatory restraint) of GKNs in vivo. Together, these data link the coordinate expression of GKNs to a glucocorticoid-responsive and likely shared transcriptional enhancer mechanism, with its compromised activation contributing to dual GKN loss during GC progression.NEW & NOTEWORTHY Gastrokine 2 (GKN2) is an anti-inflammatory protein produced by the gastric epithelium. GKN2 expression is progressively lost during gastric cancer (GC), which is believed to play a casual role in GC development. Here, we use bacterial artificial chromosome transgenic studies to identify a glucocorticoid-responsive enhancer element that likely governs expression of GKN1/GKN2, which, via parallel expression loss of the anti-inflammatory glucocorticoid receptor, reveals a novel mechanism to explain the loss of GKN2 during GC pathogenesis.

Differential therapeutic effects of PARP and ATR inhibition combined with radiotherapy in the treatment of subcutaneous versus orthotopic lung tumour models.

BACKGROUND: Subcutaneous mouse tumour models are widely used for the screening of novel antitumour treatments, although these models are poor surrogate models of human cancers. METHODS: We compared the antitumour efficacy of the combination of ionising radiation (IR) with two DNA damage response inhibitors, the PARP inhibitor olaparib and the ATR inhibitor AZD6738 (ceralasertib), in subcutaneous versus orthotopic cancer models. RESULTS: Olaparib delayed the growth of irradiated Lewis lung carcinoma (LL2) subcutaneous tumours, in agreement with previous reports in human cell lines. However, the olaparib plus IR combination showed a very narrow therapeutic window against LL2 lung orthotopic tumours, with nearly no additional antitumour effect compared with that of IR alone, and tolerability issues emerged at high doses. The addition of AZD6738 greatly enhanced the efficacy of the olaparib plus IR combination treatment against subcutaneous but not orthotopic LL2 tumours. Moreover, olaparib plus AZD6738 administration concomitant with IR even worsened the response to radiation of head and neck orthotopic tumours and induced mucositis. CONCLUSIONS: These major differences in the responses to treatments between subcutaneous and orthotopic models highlight the importance of using more pathologically relevant models, such as syngeneic orthotopic models, to determine the most appropriate therapeutic approaches for translation to the clinic.

Near-Infrared Dye-Labeled Anti-Prostate Stem Cell Antigen Minibody Enables Real-Time Fluorescence Imaging and Targeted Surgery in Translational Mouse Models.

PURPOSE: The inability to intraoperatively distinguish primary tumor, as well as lymphatic spread, increases the probability of positive surgical margins, tumor recurrence, and surgical toxicity. The goal of this study was to develop a tumor-specific optical probe for real-time fluorescence-guided surgery. EXPERIMENTAL DESIGN: A humanized antibody fragment against PSCA (A11 minibody, A11 Mb) was conjugated with a near-infrared fluorophore, IRDye800CW. The integrity and binding of the probe to PSCA were confirmed by gel electrophoresis, size-exclusion chromatography, and flow cytometry, respectively. The ability of the probe to detect tumor-infiltrated lymph nodes and metastatic lesions was evaluated in 2 xenograft models, as well as in transgenic mice expressing human PSCA (hPSCA). An invasive intramuscular model was utilized to evaluate the efficacy of the A11 Mb-IRDye800CW-guided surgery. RESULTS: A11 Mb was successfully conjugated with IRDye800CW and retained specific binding to PSCA. In vivo imaging showed maximal signal-to-background ratios at 48 hours. The A11 Mb-IRDye800CW specifically detected PSCA-positive primary tumors, tumor-infiltrated lymph nodes, and distant metastases with high contrast. Fluorescence guidance facilitated more complete tumor resection, reduced tumor recurrence, and improved overall survival, compared with conventional white light surgery. The probe successfully identified primary orthotopic tumors and metastatic lesions in hPSCA transgenic mice. CONCLUSIONS: Real-time fluorescence image-guided surgery with A11 Mb-IRDye800CW enabled detection of lymph node metastases and positive surgical margins, facilitated more complete tumor removal, and improved survival, compared with white light surgery. These results may be translatable into clinical practice to improve surgical and patient outcomes.

Fall in thyroid stimulating hormone (TSH) may be an early marker of ipilimumab-induced hypophysitis.

PURPOSE: Hypophysitis develops in up to 19% of melanoma patients treated with ipilimumab, a cytotoxic T-lymphocyte antigen-4 antibody. Early detection may avert life-threatening hypopituitarism. We aimed to assess the incidence of ipilimumab-induced hypophysitis (IH) at a quaternary melanoma referral centre, and to determine whether cortisol or thyroid stimulating hormone (TSH) monitoring could predict IH onset. METHODS: We performed a retrospective cohort study of ipilimumab-treated patients at a quaternary melanoma referral centre in Australia. The inclusion criteria were patients with metastatic or unresectable melanoma treated with ipilimumab monotherapy, and cortisol and TSH measurements prior to ≥ 2 infusions. The main outcomes were IH incidence and TSH and cortisol patterns in patients who did and did not develop IH. RESULTS: Of 78 ipilimumab-treated patients, 46 met the study criteria and 9/46 (20%) developed IH at a median duration of 13.0 weeks (range 7.7-18.1) following ipilimumab initiation. All patients whose TSH fell ≥ 80% compared to baseline developed IH, and, in 5/9 patients with IH, TSH fell prior to cortisol fall and IH diagnosis. Pre-cycle-4 TSH was significantly lower in those who developed IH (0.31 vs. 1.73 mIU/L, P = 0.006). TSH fall was detected at a median time of 9.2 (range 7.7-16.4) weeks after commencing ipilimumab, and a median of 3.6 (range of - 1.4 to 9.7) weeks before IH diagnosis. There was no difference in TSH between the groups before cycles 1-3 or in cortisol before cycles 1-4. CONCLUSIONS: TSH fall ≥ 80% may be an early marker of IH. Serial TSH measurement during ipilimumab therapy may be an inexpensive tool to expedite IH diagnosis.

Nutrition assessment and its relationship with performance and Glasgow prognostic scores in Vietnamese patients with esophageal cancer.

BACKGROUND AND OBJECTIVES: To determine the nutritional status of patients with esophageal cancer, and to investigate its relationship with performance status and prognosis. METHODS AND STUDY DESIGN: This clinical, cross-sectional study was conducted from August 2014 to February 2015 at National Cancer Hospital, Hanoi, Vietnam. Stage III/IV esophageal cancer patients were assessed for their nutritional status (patient-generated subjective global assessment (PG-SGA) and SGA scores, BMI, mid-arm circumference (MAC), energy and protein intakes, weight changes, Karnofsky and Eastern cooperative oncology group performance scores (KPS/ECOG), and Glasgow prognostic score (GPS). RESULTS: Sixty-four male patients were enrolled. The mean ± standard deviation of PG-SGA score was 9.88±4.41. SGA revealed 44% as class B and 6.2% as class C. The BMI revealed 43.8% of patients were underweight. MAC measurement revealed 29.7% of undernourished patients. Patients with an energy intake <25 kcal/kg/d comprised 54.7%, and 48.4% with <1 g/kg/day of protein. Totally, 68.8%, 84.4% and 92.2% patients exhibited weight loss past 2-weeks, one-month and six-months, respectively. The PG-SGA and SGA strongly correlated with the KPS (r=-0.717 and 0.632, both p<0.001) and ECOG (r=0.672 and 0.626, both p<0.001), but were weakly correlated with the GPS (r=0.332 and 0.278, p<0.01 and 0.05). The KPS, ECOG, BMI, MAC, energy and protein intakes, and weight change were not correlated with the GPS. CONCLUSIONS: Malnutrition, weight change, and insufficient intake were noteworthy in esophageal cancer patients. The PG-SGA and SGA were strongly correlated with the performance status, but weakly correlated with prognostic indices.

Risk factors for high-risk and multi-type Human Papillomavirus infections among women in Ho Chi Minh City, Vietnam: a cross-sectional study.

BACKGROUND: Concurrent infection with multiple types of Human Papillomavirus (HPV) is associated with an increased risk of cervical cancer; yet, little is known about risk factors for concurrent HPV infection in Vietnam. This study investigated the prevalence of and risk factors for high-risk-type HPV and multi-type HPV infections among women in Ho Chi Minh City, Vietnam. METHODS: Data were collected from a population-based survey of 1,550 women (mean age = 42.4; SD = 9.5), using a multi-stage sampling process. Socio-demographic and behavioral variables were obtained by self-report. HPV genotypes in cervical specimens were identified using PCR protocols. RESULTS: The prevalence of any high-risk HPV infection was 9.0%, and of multi-type HPV infection was 1.9%. In the HPV+ subsample, the percentage of high-risk HPV was 84% and of multi-type HPV was 20%. All multi-type HPV infections were high-risk-type. Lifetime smoking and older age of first sex were significantly associated with any high-risk and multi-type HPV infections. Regular condom use was inversely associated with high-risk and multi-type HPV infection. CONCLUSIONS: Risk factors for high-risk and multi-type HPV infections were similar. Further research and intervention are needed to reduce HPV infections in order to prevent HPV-related cancers.

Primitive macrophages control HSPC mobilization and definitive haematopoiesis.

In vertebrates, haematopoietic stem/progenitor cells (HSPCs) first emerge in the aorta-gonad-mesonephros (AGM) before colonizing transitory and subsequently definitive haematopoietic organs allowing haematopoiesis throughout adult life. Here we identify an unexpected primitive macrophage population accumulated in the dorsal mesenteric mesoderm surrounding the dorsal aorta of the human embryo and study its function in the transparent zebrafish embryo. Our study reveals dynamic interactions occurring between the HSPCs and primitive macrophages in the AGM. Specific chemical and inducible genetic depletion of macrophages or inhibition of matrix metalloproteinases (Mmps) leads to an accumulation of HSPCs in the AGM and a decrease in the colonization of haematopoietic organs. Finally, in vivo zymography demonstrates the function of primitive macrophages in extracellular matrix degradation, which allows HSPC migration through the AGM stroma, their intravasation, leading to the colonization of haematopoietic organs and the establishment of definitive haematopoiesis.

Prevalence of malnutrition in patients admitted to a major urban tertiary care hospital in Hanoi, Vietnam.

OBJECTIVES: To determine the prevalence of malnutrition using anthropometric measures among hospitalized pediatric and adult patients admitted at Bach Mai Hospital, Hanoi, Vietnam. METHODS: A one-day cross-sectional survey was used in selected wards (Pediatrics, Surgery, Intensive Care Unit, Renal Diseases, Gastroenterology Diseases, Respiratory Diseases, and Endocrinology). Unavailable patients and those discharged within 24 hours were excluded. Anthropometric data included body weight, height (or length), and mid-upper arm circumference. The type, severity, and prevalence rate of malnutrition were defined based on World Health Organization (WHO) criteria. RESULTS: The sample was hospitalized children and adults: 108 and 571 were children aged 6 months to 18.9 years old and adult patients, respectively. The overall rate of pediatric wasting (weight-for-height ≤ -2 SD or BMI ≤ -2 SD, kg/m²) was 19.0% (n= 19/100) and that of stunting (height-for-age ≤ -2 SD) was 13.9% (n=14/101). Using either the mid-upper arm circumference <11.5 cm or the weight-for-height and weight-for-length ≤ -3 SD, the rate of severe wasting among children aged 6-59 months old was 7.0% (n=3/43). None of the children were obese based on weight-for-length, weight-for-height, or BMI. In adults, the prevalence of under-nutrition (BMI<18.5 kg/m²) was 33.3% (n=141/423) while that of obesity (BMI ≥ 30 kg/m²) was 0.9% (n=4/423). Adults admitted to the Respiratory Diseases ward had the highest prevalence of under-nutrition, 40.9% (n=38/93). CONCLUSIONS: The prevalence of malnutrition was high in this cohort of hospitalized patients, particularly in adults, but comparable to other published reports. Obesity was nearly nonexistent in both children and adults.

Economic evaluation of cell salvage in pediatric surgery.

BACKGROUND: Red blood cells are a scarce resource with demand outstripping supply. Use of intraoperative red cell salvage (CS) - the process of collecting shed blood during surgery and reinfusing it to patients - is often used as an effective blood conservation strategy. However, little is known about the economic impact of CS during pediatric surgery. METHODS: A decision tree model was used to estimate the transfusion-related costs per patient (2010 USD) from a healthcare system perspective of four transfusion strategies among children undergoing elective orthopedic or cardiac surgery: (i) CS followed by allogeneic transfusion, (ii) CS followed by autologous transfusion, (iii) allogeneic transfusion alone, and (iv) autologous transfusion alone. RESULTS: Cell salvage and allogeneic transfusion was the least expensive strategy (USD 883.3) followed by CS and autologous blood transfusion (USD 1,269.7), allogeneic transfusion alone (USD 1,443.0), and autologous transfusion alone (USD 1,824.7). Savings associated with CS use persisted in separate analyses of orthopedic and cardiac surgery, as well as in one-way and probabilistic sensitivity analyses. CONCLUSIONS: Use of CS, particularly along with allogeneic blood transfusion, appears cost-saving and cost-effective in pediatric surgery.

Impact of a conservative red blood cell transfusion strategy in children undergoing hematopoietic stem cell transplantation.

A 2008 randomized trial of critically ill, but stable, children reported the safety of transfusing red blood cells at a hemoglobin threshold of 7 g/dL. In 2009, we adopted the same transfusion criteria in our hematopoietic stem cell transplantation patients. Regression modeling was used to compare data obtained during primary admission for hematopoietic stem cell transplantation in calendar years before and after our practice change. Sixty-six patients admitted in the preintervention year were compared with 75 postintervention. Pre- and postpatients were similar in diagnoses and type of transplantations. Postintervention, median hemoglobin pretransfusion significantly decreased from 8.8 g/dL to 6.8 g/dL (P < .0001). In addition, transfused red blood cell units received by patients dropped from 4 (interquartile range [IQR] 3, 8) to 3 (IQR, 2, 5), (P = .002), and number of transfusion days per patients decreased from 4 (IQR, 2,5) to 3 (IQR, 2, 5), (P = .01). There were no differences in length of stay, time to engraftment, or 100-day mortality. Median blood product charges per patient significantly decreased ($3,624 [IQR, $2,265, $6,040] to $2,185 [IQR, $1,812, $3,997], P = .004). Our initial experience suggests that implementation of a conservative transfusion strategy in otherwise stable children undergoing hematopoietic stem cell transplantation appears safe and lowers transfusion exposures.