FORMULATION AND PHYSICAL CHARACTERIZATION OF CURCUMIN NANOPARTICLE TRANSDERMAL PATCH

Objective: This study was conducted to formulate curcumin nanoparticles transdermal patches and to evaluate their physical characterization. Methods: Curcumin nanoparticles transdermal patches were formulated by the casting evaporation method. Transdermal patches were made using combinations of hydroxypropyl methylcellulose (HPMC) and ethyl cellulose (EC) at ratios of 4.5:1.5 for Formula 1 (F1), 4:1 for Formula 2 (F2), 3.5:1.5 for Formula 3 (F3), 3:2 for Formula 4 (F4), and 2.5:2.5 for Formula 5 (F5). Physical characterization evaluation (organoleptic properties, pH, weight uniformity, thickness uniformity, percent moisture content, and tensile strength) was then performed. The permeation of curcumin nanoparticles into the skin was evaluated using Franz diffusion cells. Results: Curcumin nanoparticles transdermal patches could be formulated by the casting evaporation method with the organoleptic properties characterized as smooth, dry, yellow in color, having menthol odor, and transparent. The pH values ranged between 5.0 and 6.0. The thickness of the patches ranged from 0.1 to 0.2 mm. The average of the patches’ weight was 0.7 g, and the percent moisture content ranged from 1.0 to 6.0%. The tensile strength values were 1.0 to 2.0 N/mm. Curcumin nanoparticles could penetrate into the skin with flux values being 1.271 µg. cm-2 (F1), 0.938 µg. cm-2 (F2), 0.775 µg. cm-2 (F3), 0.837 µg. cm-2 (F4), and 0.569 µg. cm-2 (F5). Conclusion: All patches met the requirement of the physical characterization for the transdermal patch

Accumulation of copy number alterations and clinical progression across advanced prostate cancer

Background: Genomic copy number alterations commonly occur in prostate cancer and are one measure of genomic instability. The clinical implication of copy number change in advanced prostate cancer, which defines a wide spectrum of disease from high-risk localised to metastatic, is unknown. Methods: We performed copy number profiling on 688 tumour regions from 300 patients, who presented with advanced prostate cancer prior to the start of long-term androgen deprivation therapy (ADT), in the control arm of the prospective randomised STAMPEDE trial. Patients were categorised into metastatic states as follows; high-risk non-metastatic with or without local lymph node involvement, or metastatic low/high volume. We followed up patients for a median of 7 years. Univariable and multivariable Cox survival models were fitted to estimate the association between the burden of copy number alteration as a continuous variable and the hazard of death or disease progression. Results: The burden of copy number alterations positively associated with radiologically evident distant metastases at diagnosis (P=0.00006) and showed a non-linear relationship with clinical outcome on univariable and multivariable analysis, characterised by a sharp increase in the relative risk of progression (P=0.003) and death (P=0.045) for each unit increase, stabilising into more modest increases with higher copy number burdens. This association between copy number burden and outcome was similar in each metastatic state. Copy number loss occurred significantly more frequently than gain at the lowest copy number burden quartile (q=4.1 × 10−6). Loss of segments in chromosome 5q21-22 and gains at 8q21-24, respectively including CHD1 and cMYC occurred more frequently in cases with higher copy number alteration (for either region: Kolmogorov–Smirnov distance, 0.5; adjusted P<0.0001). Copy number alterations showed variability across tumour regions in the same prostate. This variance associated with increased risk of distant metastases (Kruskal-Wallis test P=0.037). Conclusions: Copy number alteration in advanced prostate cancer associates with increased risk of metastases at diagnosis. Accumulation of a limited number of copy number alterations associates with most of the increased risk of disease progression and death. The increased likelihood of involvement of specific segments in high copy number alteration burden cancers may suggest an order underlying the accumulation of copy number changes

Abiraterone acetate and prednisolone with or without enzalutamide for high-risk non-metastatic prostate cancer: a meta-analysis of primary results from two randomised controlled phase 3 trials of the STAMPEDE platform protocol

© 2022 The Authors. Published by Elsevier. This is an open access article available under a Creative Commons licence. The published version can be accessed at the following link on the publisher’s website: https://www.thelancet.com/journals/lancet/article/PIIS0140-6736(21)02437-5/fulltextBackground Men with high-risk non-metastatic prostate cancer are treated with androgen-deprivation therapy (ADT) for 3 years, often combined with radiotherapy. We analysed new data from two randomised controlled phase 3 trials done in a multiarm, multistage platform protocol to assess the efficacy of adding abiraterone and prednisolone alone or with enzalutamide to ADT in this patient population. Methods These open-label, phase 3 trials were done at 113 sites in the UK and Switzerland. Eligible patients (no age restrictions) had high-risk (defined as node positive or, if node negative, having at least two of the following: tumour stage T3 or T4, Gleason sum score of 8–10, and prostate-specific antigen [PSA] concentration ≥40 ng/mL) or relapsing with high-risk features (≤12 months of total ADT with an interval of ≥12 months without treatment and PSA concentration ≥4 ng/mL with a doubling time of <6 months, or a PSA concentration ≥20 ng/mL, or nodal relapse) non-metastatic prostate cancer, and a WHO performance status of 0–2. Local radiotherapy (as per local guidelines, 74 Gy in 37 fractions to the prostate and seminal vesicles or the equivalent using hypofractionated schedules) was mandated for node negative and encouraged for node positive disease. In both trials, patients were randomly assigned (1:1), by use of a computerised algorithm, to ADT alone (control group), which could include surgery and luteinising-hormone-releasing hormone agonists and antagonists, or with oral abiraterone acetate (1000 mg daily) and oral prednisolone (5 mg daily; combination-therapy group). In the second trial with no overlapping controls, the combination-therapy group also received enzalutamide (160 mg daily orally). ADT was given for 3 years and combination therapy for 2 years, except if local radiotherapy was omitted when treatment could be delivered until progression. In this primary analysis, we used meta-analysis methods to pool events from both trials. The primary endpoint of this meta-analysis was metastasis-free survival. Secondary endpoints were overall survival, prostate cancer-specific survival, biochemical failure-free survival, progression-free survival, and toxicity and adverse events. For 90% power and a one-sided type 1 error rate set to 1·25% to detect a target hazard ratio for improvement in metastasis-free survival of 0·75, approximately 315 metastasis-free survival events in the control groups was required. Efficacy was assessed in the intention-to-treat population and safety according to the treatment started within randomised allocation. STAMPEDE is registered with ClinicalTrials.gov, NCT00268476, and with the ISRCTN registry, ISRCTN78818544. Findings Between Nov 15, 2011, and March 31, 2016, 1974 patients were randomly assigned to treatment. The first trial allocated 455 to the control group and 459 to combination therapy, and the second trial, which included enzalutamide, allocated 533 to the control group and 527 to combination therapy. Median age across all groups was 68 years (IQR 63–73) and median PSA 34 ng/ml (14·7–47); 774 (39%) of 1974 patients were node positive, and 1684 (85%) were planned to receive radiotherapy. With median follow-up of 72 months (60–84), there were 180 metastasis-free survival events in the combination-therapy groups and 306 in the control groups. Metastasis-free survival was significantly longer in the combination-therapy groups (median not reached, IQR not evaluable [NE]–NE) than in the control groups (not reached, 97–NE; hazard ratio [HR] 0·53, 95% CI 0·44–0·64, p<0·0001). 6-year metastasis-free survival was 82% (95% CI 79–85) in the combination-therapy group and 69% (66–72) in the control group. There was no evidence of a difference in metatasis-free survival when enzalutamide and abiraterone acetate were administered concurrently compared with abiraterone acetate alone (interaction HR 1·02, 0·70–1·50, p=0·91) and no evidence of between-trial heterogeneity (I2 p=0·90). Overall survival (median not reached [IQR NE–NE] in the combination-therapy groups vs not reached [103–NE] in the control groups; HR 0·60, 95% CI 0·48–0·73, p<0·0001), prostate cancer-specific survival (not reached [NE–NE] vs not reached [NE–NE]; 0·49, 0·37–0·65, p<0·0001), biochemical failure-free-survival (not reached [NE–NE] vs 86 months [83–NE]; 0·39, 0·33–0·47, p<0·0001), and progression-free-survival (not reached [NE–NE] vs not reached [103–NE]; 0·44, 0·36–0·54, p<0·0001) were also significantly longer in the combination-therapy groups than in the control groups. Adverse events grade 3 or higher during the first 24 months were, respectively, reported in 169 (37%) of 451 patients and 130 (29%) of 455 patients in the combination-therapy and control groups of the abiraterone trial, respectively, and 298 (58%) of 513 patients and 172 (32%) of 533 patients of the combination-therapy and control groups of the abiraterone and enzalutamide trial, respectively. The two most common events more frequent in the combination-therapy groups were hypertension (abiraterone trial: 23 (5%) in the combination-therapy group and six (1%) in control group; abiraterone and enzalutamide trial: 73 (14%) and eight (2%), respectively) and alanine transaminitis (abiraterone trial: 25 (6%) in the combination-therapy group and one (<1%) in control group; abiraterone and enzalutamide trial: 69 (13%) and four (1%), respectively). Seven grade 5 adverse events were reported: none in the control groups, three in the abiraterone acetate and prednisolone group (one event each of rectal adenocarcinoma, pulmonary haemorrhage, and a respiratory disorder), and four in the abiraterone acetate and prednisolone with enzalutamide group (two events each of septic shock and sudden death). Interpretation Among men with high-risk non-metastatic prostate cancer, combination therapy is associated with significantly higher rates of metastasis-free survival compared with ADT alone. Abiraterone acetate with prednisolone should be considered a new standard treatment for this population.Cancer Research UK, UK Medical Research Council, Swiss Group for Clinical Cancer Research, Janssen, and Astellas.Published versio

Implementasi Keadilan Gender di Pondok Pesantren Sabilurrosyad Kota Malang

Gender equality still becomes a sensitive issue in Islamic educational institutions, especially in pesantren, a traditional Islamic educational institution. There is a strong assumption that in the socio-religious tradition of pesantren, women's subordination still practices widely. However, some pesatren take serious attention to overcome this issue through their daily activities within pesantren. One of which is Pesantren Sabilurrosyad in Malang, East Java. This article aims to elaborate on the realization of gender justice in the pesantren. This research uses a qualitative approach through observation and in-depth interviews with the board members of pesantren, as well as its male and female students. The results showed that the Pesantren Sabilurrosyad had implemented the values of gender justice in their socio-religious activities. The implementation forms include providing opportunities for female students to become head of student association; female students are given freedom to recite the Koran directly to the kyai; and female teachers (ustadzah) are given the opportunity to share in one forum with male students.Kesetaraan gender masih menjadi isu sensitive di lembaga pendidikan Islam, khususnya di pondok pesantren. Terdapat anggapan bahwa dalam tradisi sosial-keagamaan di pesantren subordinasi perempuan masih terjadi. Hal ini menjadi perhatian bagi beberapa pesantren yang ingin menjadikan isu ini sebagai bagian dari aktivitas pesantren, salah satunya adalah Pondok Pesantren Sabilurrosyad di Malang. Artikel ini bertujuan untuk mengelaborasi perwujudan keadilan gender di pondok tersebut. Penelitian ini menggunakan pendekatan kualitatif melalui observasi dan wawancara mendalam terhadap para pengurus ponpes dan santri putra dan santri putri. Hasil penelitian menunjukkan bahwa pondok pesantren Sabilurrosyad telah mengimplementasikan nilai-nilai keadilan gender dalam tradisi sosial-keagamaan pesantren. Wujud implementasi tersebut diantaranya adalah memberikan kesempatan kepada santri putri untuk menjadi ketua pondok, santri putri diberikan kebebasan untuk mengaji langsung kepada kyai, serta pengajar putri (ustadzah) diberikan peluang untuk bersama 1 (satu) forum dengan santri putra

HIV-1 virological synapse formation enhances infection spread by dysregulating Aurora Kinase B.

HIV-1 spreads efficiently through direct cell-to-cell transmission at virological synapses (VSs) formed by interactions between HIV-1 envelope proteins (Env) on the surface of infected cells and CD4 receptors on uninfected target cells. Env-CD4 interactions bring the infected and uninfected cellular membranes into close proximity and induce transport of viral and cellular factors to the VS for efficient virion assembly and HIV-1 transmission. Using novel, cell-specific stable isotope labeling and quantitative mass spectrometric proteomics, we identified extensive changes in the levels and phosphorylation states of proteins in HIV-1 infected producer cells upon mixing with CD4+ target cells under conditions inducing VS formation. These coculture-induced alterations involved multiple cellular pathways including transcription, TCR signaling and, unexpectedly, cell cycle regulation, and were dominated by Env-dependent responses. We confirmed the proteomic results using inhibitors targeting regulatory kinases and phosphatases in selected pathways identified by our proteomic analysis. Strikingly, inhibiting the key mitotic regulator Aurora kinase B (AURKB) in HIV-1 infected cells significantly increased HIV activity in cell-to-cell fusion and transmission but had little effect on cell-free infection. Consistent with this, we found that AURKB regulates the fusogenic activity of HIV-1 Env. In the Jurkat T cell line and primary T cells, HIV-1 Env:CD4 interaction also dramatically induced cell cycle-independent AURKB relocalization to the centromere, and this signaling required the long (150 aa) cytoplasmic C-terminal domain (CTD) of Env. These results imply that cytoplasmic/plasma membrane AURKB restricts HIV-1 envelope fusion, and that this restriction is overcome by Env CTD-induced AURKB relocalization. Taken together, our data reveal a new signaling pathway regulating HIV-1 cell-to-cell transmission and potential new avenues for therapeutic intervention through targeting the Env CTD and AURKB activity

Western blot of cell mixing conditions.

(A) Jurkat producer cells and uninfected SupT1 target cells were mixed for 0, 5 and 60 minutes. Cells were lysed, separated by SDS-PAGE and immunoblotted with the indicated antibodies. Consistent with previous studies and our mass spec results, no changes were observed in viral Env or gag or in total cellular actin or MAPK14. Phospho-MAPK14 was unchanged 5 min after mixing and decreased by 60 min. In contrast, p-AKT increased at 5 and 60 min post mixing. (B) Quantitation of p-AKT and p-MAPK14. The data shown are the average mean values obtained in an experiment performed with quadruplicate samples and are representative of three independent experiments. Error bars indicate the standard deviation of the data. P-values were calculated using a standard Student’s t-test. (EPS)</p

Decision tree used to identify proteins involved in HIV cell to cell spread.

Initial screening was performed using producer cell mixing with TZM-BL target cells (Fig 4A). Potential hits were confirmed by further analysis as shown. Compounds were rejected if either producer or target cells viability was negatively affected by chemical treatment (Fig 4B). Next, potential hits were rejected if chemical treatment inhibited cell free virion infection of target cells (Fig 4D). Finally, if inhibitors enhanced syncytia formation, they were classified as a specific class of hit that affected the fusogenic activity of Env and selected for further study (Fig 5). (EPS)</p

AURKB regulation of HIV spread through virological synapse requires the C-terminal domain of Env.

Schematic of HIV-1 virological synapse. Cytoplasmic AURKB exerts an unknown negative effect on HIV-1 spread through the virological synapse which reduces the fusion activity of the HIV-1 Env protein. HIV-1 overcomes this through the CTD of HIV-1 Env which induces premature nuclear localization of AURKB to the CPC.</p

Producer cell phosphopeptides identified after VS formation.

Phosphopeptides identified by protein group, UniProt ID, representative name, and phosphorylation isoform. Columns show log2 fold changes and q-values in phosphopeptide level in experiments 1 and 2 at 5 and 60 minutes relative to 0 min controls. True in the Significant column means the phosphopeptide met criteria of a q-value ≤ 0.1 and a fold change ≥ 1.5 in magnitude in at least one experiment and time point. (XLSX)</p