Efficacy and safety of BVAC-C in HPV type 16- or 18-positive cervical carcinoma who failed 1st platinum-based chemotherapy: a phase I/IIa study.

Background: BVAC-C, a B cell- and monocyte-based immunotherapeutic vaccine transfected with recombinant HPV E6/E7, was well tolerated in HPV-positive recurrent cervical carcinoma patients in a phase I study. This phase IIa study investigates the antitumor activity of BVAC-C in patients with HPV 16- or 18-positive cervical cancer who had experienced recurrence after a platinum-based combination chemotherapy. Patients and methods: Patients were allocated to 3 arms; Arm 1, BVAC-C injection at 0, 4, 8 weeks; Arm 2, BVAC-C injection at 0, 4, 8, 12 weeks; Arm 3, BVAC-C injection at 0, 4, 8, 12 weeks with topotecan at 2, 6, 10, 14 weeks. Primary endpoints were safety and objective response rate (ORR) as assessed by an independent radiologist according to Response Evaluation Criteria in Solid Tumors version 1.1. Secondary endpoints included the disease control rate (DCR), duration of response (DOR), progression-free survival (PFS), and overall survival (OS). Results: Of the 30 patients available for analysis, the ORR was 19.2% (Arm 1: 20.0% (3/15), Arm 2: 33.3% (2/6), Arm3: 0%) and the DCR was 53.8% (Arm 1: 57.1%, Arm 2: 28.6%, Arm3: 14.3%). The median DOR was 7.5 months (95% CI 7.1-not reported), the median PFS was 5.8 months (95% CI 4.2-10.3), and the median OS was 17.7 months (95% CI 12.0-not reported). All evaluated patients showed not only inflammatory cytokine responses (IFN-γ or TNF-α) but also potent E6/E7-specific T cell responses upon vaccinations. Immune responses of patients after vaccination were correlated with their clinical responses. Conclusion: BVAC-C represents a promising treatment option and a manageable safety profile in the second-line setting for this patient population. Further studies are needed to identify potential biomarkers of response. Clinical trial registration: ClinicalTrials.gov, identifier NCT02866006.

First-in-Human Phase 1 Study of a B Cell- and Monocyte-Based Immunotherapeutic Vaccine against HER2-Positive Advanced Gastric Cancer.

PURPOSE: BVAC-B is an autologous B cell- and monocyte-based immunotherapeutic vaccine that contains cells transfected with a recombinant human epidermal growth factor receptor 2 (HER2) gene and loaded with the natural killer T cell ligand alpha-galactosylceramide. Here, we report the first BVAC-B study in patients with HER2-positive advanced gastric cancer. MATERIALS AND METHODS: Patients with advanced gastric cancer refractory to standard treatment with HER2+ immunohistochemistry ≥ 1 were eligible for treatment. Patients were administered low (2.5×107 cells/dose), medium (5.0×107 cells/dose), or high dose (1.0×108 cells/dose) of BVAC-B intravenously four times every 4 weeks. Primary endpoints included safety and maximum tolerated BVAC-B dose. Secondary endpoints included preliminary clinical efficacy and BVAC-B-induced immune responses. RESULTS: Eight patients were treated with BVAC-B at low (n=1), medium (n=1), and high doses (n=6). No dose-limiting toxicity was observed, while treatment-related adverse events (TRAEs) were observed in patients treated with medium and high doses. The most common TRAEs were grade 1 (n=2) and grade 2 (n=2) fever. Out of the six patients treated with high-dose BVAC-B, three had stable disease with no response. Interferon gamma, tumor necrosis factor-α, and interleukin-6 increased after BVAC-B treatment in all patients with medium and high dose, and HER2-specific antibody was detected in some patients. CONCLUSION: BVAC-B monotherapy had a safe toxicity profile with limited clinical activity; however, it activated immune cells in heavily pretreated patients with HER2-positive gastric cancer. Earlier treatment with BVAC-B and combination therapy is warranted for evaluation of clinical efficacy.

The Chimeric Adenovirus (Ad5/35) Expressing Engineered Spike Protein Confers Immunity against SARS-CoV-2 in Mice and Non-Human Primates.

Several COVID-19 platforms have been licensed across the world thus far, but vaccine platform research that can lead to effective antigen delivery is still ongoing. Here, we constructed AdCLD-CoV19 that could modulate humoral immunity by harboring SARS-CoV-2 antigens onto a chimeric adenovirus 5/35 platform that was effective in cellular immunity. By replacing the S1/S2 furin cleavage sequence of the SARS-CoV-2 Spike (S) protein mounted on AdCLD-CoV19 with the linker sequence, high antigen expression was confirmed in various cell lines. The high levels of antigen expression contributed to antigen-specific antibody activity in mice and non-human primates (NHPs) with a single vaccination of AdCLD-CoV19. Furthermore, the adenovirus-induced Th1 immune response was specifically raised for the S protein, and these immune responses protected the NHP against live viruses. While AdCLD-CoV19 maintained neutralizing antibody activity against various SARS-CoV-2 variants, it was reduced to single vaccination for ß and ο variants, and the reduced neutralizing antibody activity was restored with booster shots. Hence, AdCLD-CoV19 can prevent SARS-CoV-2 with a single vaccination, and the new vaccine administration strategy that responds to various variants can maintain the efficacy of the vaccine.

Response of Scenedesmus quadricauda (Chlorophyceae) to Salt Stress Considering Nutrient Enrichment and Intracellular Proline Accumulation.

Aquatic organisms are exposed to a wide range of salinity, which could critically affect their survival and growth. However, their survival and growth response to salinity stress remain unclear. This study evaluates the growth response and intracellular proline accumulation of green algae, Scenedesmus quadricauda, isolated from brackish water, against dissolved salts stress with N and P enrichment. We tested a hypothesis that nutrient enrichment can relieve the dissolved salts stress of algae by accumulating intracellular proline, thereby improving survival and growth. Four levels of salinity (0, 3, 6, 12 psu) were experimentally manipulated with four levels of nutrient stoichiometry (N:P ratio = 2, 5, 10, 20) at constant N (1 mgN/L) or P levels (0.05 and 0.5 mgP/L). In each set of experiments, growth rate and intracellular proline content were measured in triplicate. The highest level of salinity inhibited the growth rate of S. quadricauda, regardless of the nutrient levels. However, with nutrient enrichment, the alga showed tolerance to dissolved salts, reflecting intracellular proline synthesis. Proline accumulation was most prominent at the highest salinity level, and its maximum value appeared at the highest N:P ratio (i.e., highest N level) in all salinity treatments, regardless of P levels. Therefore, the effects of P and N on algal response to salt stress differ.

Phase I Study of a B Cell-Based and Monocyte-Based Immunotherapeutic Vaccine, BVAC-C in Human Papillomavirus Type 16- or 18-Positive Recurrent Cervical Cancer.

: BVAC-C is a B cell-based and monocyte-based immuno-therapeutic vaccine transfected with a recombinant human papillomavirus (HPV) 16/18 E6/E7 gene and loaded with alpha-galactosyl ceramide, which is a natural killer T cell ligand. This phase I study sought to determine the tolerability and immunogenicity of BVAC-C in platinum-resistant recurrent cervical cancer patients. Patients with HPV 16-positive or 18-positive recurrent or persistent cervical cancer who had received at least one prior platinum-based combination chemotherapy were enrolled. BVAC-C was injected intravenously three times every four weeks, and dose escalation was planned in a three-patient cohort design at doses of 1 × 107, 4 × 107, or 1 × 108 cells/dose. Eleven patients were enrolled, and six (55%) patients had received two or more lines of platinum-based chemotherapy prior to enrollment. Treatment-related adverse events (TRAEs) were observed in 21 cycles. Most TRAEs were mild fever (n = 6, 55%) or myalgia (n = 4, 36%). No dose-limiting toxicities occurred. The overall response rate was 11% among nine patients evaluable, and the duration of response was 10 months. Five patients (56%) achieved a stable disease for 4.2-11 months as their best overall response. The median progression-free survival in all patients was 6.8 months (95% CI, 3.2 to infinite months), and the overall survival rate at 6 and 12 months was 89% (95% CI, 71 to 100%) and 65% (95% CI, 39 to 100%), respectively. BVAC-C induced the activation of natural killer T cells, natural killer cells, and HPV 16/18 E6/E7-specific T cells upon vaccination in all patients evaluated. BVAC-C was well tolerated and demonstrated a durable anti-tumor activity with an immune response in HPV 16-positive or 18-positive recurrent cervical carcinoma patients. A Phase 2 efficacy trial is currently underway.

Akinete germination chamber: An experimental device for cyanobacterial akinete germination and plankton emergence.

Understanding how algal resting cells (e.g. akinetes) germinate and what factors influence their germination rate is crucial for elucidating the development of algal blooms and their succession. While laboratory studies have demonstrated algal germination rate and some key factors affecting the germination, the use of artificially induced akinetes and/or removal of the sediments are obviously limiting in simulating the natural environment when designing such controlled experiments. This study introduce a laboratory Akinete Germination Chamber (AGC) that facilitates research for cyanobacterial akinete germination and emergence in an environment similar to natural conditions while minimizing sediment disturbance. The fundamental difference between AGC method and the conventional microplate method is that AGC incorporates the substrate from the natural environment whereas the microplate method does not employ sediment. Therefore, authors of this study assume that the characteristics of akinete germination between the two methods differ because the sediment influences the germination environment. The present study developed the AGC method as an efficient tool to understand harmful cyanobacterial bloom formation. For validation of the AGC method, this study evaluated akinete germination of Dolichospermum circinale (Anabaena circinalis) with different temperature and nutrient condition and then compared the results with those generated by conventional methods The results showed a marked difference in the maximal germination rate between two methods (78% and 35% in the AGC and the microplate, respectively; p < 0.05) at optimum germination temperature (25 °C for both the AGC and the microplate). The nutrient effect also demonstrated clear difference (p < 0.01) in the germination rate between two methods; 88%, 68% and 78% in the AGC and 15%, 20% and 15% in the microplate with -N+P, +N-P, and +N+P condition of CB medium, respectively. Importantly, both DW and -N-P treatments in the AGC induced a little germination of akinete (4.2 ±â€¯1.4% and 5.0 ±â€¯7.1%, respectively), whereas no germination was occurred in the DW treatment in the microplate, suggesting a possible positive effect of sediment on akinete germination. With these results, this study suspects that these differences were largely attributable to natural sediment. Also sediment-accompanied properties, possibly such as nutrient availability, heat budget, micronutrients, and bacteria might have some potential effects on akinete germination. The AGC method can overcome the limitations of the conventional microplate method, and that it is applicable in studies on pelagic-benthic coupling.

IL-21-mediated reversal of NK cell exhaustion facilitates anti-tumour immunity in MHC class I-deficient tumours.

During cancer immunoediting, loss of major histocompatibility complex class I (MHC-I) in neoplasm contributes to the evasion of tumours from host immune system. Recent studies have demonstrated that most natural killer (NK) cells that are found in advanced cancers are defective, releasing the malignant MHC-I-deficient tumours from NK-cell-dependent immune control. Here, we show that a natural killer T (NKT)-cell-ligand-loaded tumour-antigen expressing antigen-presenting cell (APC)-based vaccine effectively eradicates these advanced tumours. During this process, we find that the co-expression of Tim-3 and PD-1 marks functionally exhausted NK cells in advanced tumours and that MHC-I downregulation in tumours is closely associated with the induction of NK-cell exhaustion in both tumour-bearing mice and cancer patients. Furthermore, the recovery of NK-cell function by IL-21 is critical for the anti-tumour effects of the vaccine against advanced tumours. These results reveal the process involved in the induction of NK-cell dysfunction in advanced cancers and provide a guidance for the development of strategies for cancer immunotherapy.

Unpredictability of fighter pilots' g duration tolerance by anthropometric and physiological characteristics.

BACKGROUND: While the factors affecting fighter pilots' G level tolerance have been widely accepted, the factors affecting fighter pilots' G duration tolerance have not been well understood. METHODS: Thirty-eight subjects wearing anti-G suits were exposed to sustained high G forces using a centrifuge. The subjects exerted AGSM and decelerated the centrifuge when they reached the point of loss of peripheral vision. The G profile consisted of a +2.3 G onset rate, +7.3 G single plateau, and -1.6 G offset rate. Each subject's G tolerance time was recorded and the relationship between the tolerance time and the subject's anthropometric and physiological factors were analyzed. RESULTS: The mean tolerance time of the 38 subjects was 31.6 s, and the min and max tolerance times were 20 s and 58 s, respectively. The correlation analysis indicated that none of the factors had statistically significant correlations with the subjects' G duration tolerance. Stepwise multiple regression analysis showed that G duration tolerance was not dependent on any personal factors of the subjects. After the values of personal factors were simplified into 0 or 1, the t-test analysis showed that subjects' heights were inversely correlated with G duration tolerance at a statistically significant level. However, a logistic regression analysis suggested that the effect of the height factor to a pilot's G duration tolerance was too weak to be used as a predictor of a pilot's G tolerance. CONCLUSION: Fighter pilots' G duration tolerance could not be predicted by pilots' anthropometric and physiological factors.

DISCRIMINATIVE POWER OF NUCLEAR rDNA SEQUENCES FOR THE DNA TAXONOMY OF THE DINOFLAGELLATE GENUS PERIDINIUM (DINOPHYCEAE)(1).

The genus Peridinium Ehrenb. comprises a group of highly diversified dinoflagellates. Their morphological taxonomy has been established over the last century. Here, we examined relationships within the genus Peridinium, including Peridinium bipes F. Stein sensu lato, based on a molecular phylogeny derived from nuclear rDNA sequences. Extensive rDNA analyses of nine selected Peridinium species showed that intraspecies genetic variation was considerably low, but interspecies genetic divergence was high (>1.5% dissimilarity in the nearly complete 18S sequence; >4.4% in the 28S rDNA D1/D2). The 18S and 28S rDNA Bayesian tree topologies showed that Peridinium species grouped according to their taxonomic positions and certain morphological characters (e.g., epithecal plate formula). Of these groups, the quinquecorne group (plate formula of 3', 2a, 7″) diverged first, followed by the umbonatum group (4', 2a, 7″) and polonicum group (4', 1a, 7″). Peridinium species with a plate formula of 4', 3a, 7″ diverged last. Thus, 18S and 28S rDNA D1/D2 sequences are informative about relationships among Peridinium species. Statistical analyses revealed that the 28S rDNA D1/D2 region had a significantly higher genetic divergence than the 18S rDNA region, suggesting that the former as DNA markers may be more suitable for sequence-based delimitation of Peridinium. The rDNA sequences had sufficient discriminative power to separate P. bipes f. occultaum (Er. Lindem.) M. Lefèvre and P. bipes f. globosum Er. Lindem. into two distinct species, even though these taxa are morphologically only marginally discriminated by spines on antapical plates and the shape of red bodies during the generation of cysts. Our results suggest that 28S rDNA can be used for all Peridinium species to make species-level taxonomic distinctions, allowing improved taxonomic classification of Peridinium.

Relationship between cyanobacterial biomass and total microcystin-LR levels in drinking and recreational water.

Cyanobacterial biomass, chlorophyll-a, and microcystin-LR levels were monitored in drinking and recreational water in Seoul, South Korea and three satellite cities from Oct 2006 to Aug 2007. Total microcystin-LR was the sum of particulate and dissolved microcystin. Except during cold periods, toxic cyanobacteria, including Anabaena flos-aquae, were found at all sites. The total microcystin-LR levels were below guideline danger levels (<1.0 µg/L) except one time (1.27 µg/L in October), whereas chl-a (111.7 µg/L) and cell levels (2.6 × 105 cells/mL) were at 'vigilance' and 'alert' levels for drinking water and at 'guidance' level for recreational water, respectively. Discrepancies in these parameters may thus lead to frequent unnecessary alerts, thereby increasing water management costs.

Broad-spectrum antiviral effect of Agrimonia pilosa extract on influenza viruses.

Influenza virus continues to emerge and re-emerge, posing new threats for humans. Here we tested various Korean medicinal plant extracts for potential antiviral activity against influenza viruses. Among them, an extract of Agrimonia pilosa was shown to be highly effective against all three subtypes of human influenza viruses including H1N1 and H3N2 influenza A subtypes and influenza B virus. The EC(50) value against influenza A virus, as tested by the plaque reduction assay on MDCK cells, was 14-23 microg/ml. The extract also exhibited a virucidal effect at a concentration of 160-570 ng/ml against influenza A and B viruses when the viruses were treated with the extract prior to plaque assay. In addition, when tested in embryonated chicken eggs the extract exhibited a strong inhibitory effect in ovo on the H9N2 avian influenza virus at a concentration of 280 ng/ml. Quantitative RT-PCR analysis data showed that the extract, to some degree, suppressed viral RNA synthesis in MDCK cells. HI and inhibition of neuraminidase were observed only at high concentrations of the extract. And yet, the extract's antiviral activity required direct contact between it and the virus, suggesting that its antiviral action is mediated by the viral membrane, but does not involve the two major surface antigens, HA and NA, of the virus. The broad-spectrum antiviral activity of Agrimonia pilosa extract on various subtypes of influenza viruses merits further investigation as it may provide a means of managing avian influenza infections in poultry farms and potential avian-human transmission.

Selective inhibitory potential of silver nanoparticles on the harmful cyanobacterium Microcystis aeruginosa.

Silver nanoparticles (SNPs) at 1 mg/l inhibited the growth of the toxic cyanobacterium, Microcystis aeruginosa, by 87%. Similar results were obtained in field experiments. M. aeruginosa was more sensitive to SNPs than were green algae. SNPs may be a useful selective biocidal agent for the control of M. aeruginosa.

Molecular mechanisms of ginsenoside Rp1-mediated growth arrest and apoptosis.

Ginsenoside Rp1, a semi-synthesized ginseng saponin, was shown to have chemopreventive action and anti-metastatic potential. However, the molecular mechanisms of Rp1 on cell growth and death are not fully understood. In this study, the antiproliferative effect of Rp1 on HeLa cells in vitro was investigated. Treatment with Rp1 at 40 microM inhibited the proliferation and partial accumulation of cells at the G1 phase. Rp1-mediated G1 arrest was accompanied by decreased expression of cyclin D1, E, and A and increased expression of p21 without any significant change in p53 or phospho-p53 (Ser15). On the other hand, prolonged incubation with Rp1 at 40 microM caused apoptosis and activation of caspase-3, -8, and -9. The participation of these three caspases in apoptosis was more clearly shown in experiments using inhibitors, which markedly prevented Rp1-induced apoptosis in the case of each caspase. Cleavage of the polyADP-ribose polymerase, often used as an apoptotic marker, was also found in Rp1-induced apoptosis. Among Bcl-2 family proteins (Bad, Bax, Bid, Bcl-2), Bax and Bid were activated by Rp1 treatment, which resulted in the release of cytochrome c from mitochondria, following activation of caspase-9. These observations indicate that multiple cell cycle regulatory proteins and apoptosis-inducing proteins are regulated by Rp1 and contribute to Rp1-induced growth arrest and apoptosis.

Selective bactericidal potential of rice (Oryza sativa L. var. japonica) hull extract on Microcystis strains in comparison with green algae and zooplankton.

We examined the selective inhibitory potential of rice hull extract on the toxic cyanobacterium Microcystis aeruginosa, in comparison with inhibitory effects on two green algae (Ankistrodesmus convolutus and Scenedesmus quadricauda) and a zooplankton (Daphnia magna) species. The inhibitory effect of rice hull extract, measured by algal growth or zooplankton survival using four different concentrations of extract (1, 10, 100 and 1000 microg L(-1)), was highest on Microcystis strains (average: 98%, range: 95%-99%), followed by D. magna (average: 22%, range: 10%-47%), A. convolutus (average: 20%, range: 16%-24%), and S. quadricauda (average: 8%, range: 0%-15%). Rice hull extract had only a small effect on the growth of the green algae and Daphnia, particularly in the range 1-100 microg L(-1), and the inhibitory effect was somewhat diminished even at the 1,000 microg L(-1) level, at the end of the experimental period, especially for Daphnia. Our study indicates that rice hull extract has a strong specific algicide potential when used to combat M. aeruginosa.

Growth inhibition of Cyanobacteria by ultrasonic radiation: laboratory and enclosure studies.

The growth of Microcystis aeruginosa UTEX 2388 was repressed by ultrasonic radiation and resulted in an increased chlorophyll a content and cell size, suggesting the inhibition of cell division. However, growth was recovered immediately after the interruption of ultrasonication. In addition to the disruption of gas vesicles, other mechanisms of growth inhibition were also investigated. Although free radicals were produced by ultrasonication and hydrogen peroxide, the resulting lipid peroxidation in the cells was not comparable, indicating minimal damage by the free radicals. Ultrasonic radiation late in the day was found to be most effective in reducing the growth rate of M. aeruginosa, and this timing also corresponded to the phase of daily cell division. In an enclosure experiment, ultrasonic radiation reduced the pH, DO, total nitrogen, and total phosphorus, whereas it increased the water temperature, conductivity, and orthophosphate concentration. The algal cell density and chlorophyll a concentration drastically decreased after 3 d of ultrasonication, plus the cyanobacterial proportion was selectively reduced as compared to other algal species. Accordingly, ultrasonic radiation would appear to have considerable potential as an effective control method for cyanobacterial blooms.