Anticancer Effects of Fucoxanthin through Cell Cycle Arrest, Apoptosis Induction, and Angiogenesis Inhibition in Triple-Negative Breast Cancer Cells.

The absence of progesterone receptors, estrogen receptors, and human epidermal growth factor receptor-2 restricts the therapy choices for treating triple-negative breast cancer (TNBC). Moreover, conventional medication is not highly effective in treating TNBC, and developing effective therapeutic agents from natural bioactive compounds is a viable option. In this study, the anticancer effects of the natural compound fucoxanthin were investigated in two genetically different models of TNBC cells: MDA-MB-231 and MDA-MB-468 cells. Fucoxanthin had a significant anticancer effect in both cell lines at a concentration range of 1.56-300 µM. The compound decreased cell viability in both cell lines with higher potency in MDA-MB-468 cells. Meanwhile, proliferation assays showed similar antiproliferative effects in both cell lines after 48 h and 72 h treatment periods. Flow cytometry and Annexin V-FITC apoptosis assay revealed the ability of fucoxanthin to induce apoptosis in MDA-MB-231 only. Cell cycle arrest analysis showed that the compound also induced cell cycle arrest at the G1 phase in both cell lines, accompanied by more cell cycle arrest in MDA-MB-231 cells at S-phase and a higher cell cycle arrest in the MDA-MB-468 cells at G2-phase. Wound healing and migration assay showed that in both cell lines, fucoxanthin prevented migration, but was more effective in MDA-MB-231 cells in a shorter time. In both angiogenic cytokine array and RT-PCR studies, fucoxanthin (6.25 µM) downregulated VEGF-A and -C expression in TNF-α-stimulated (50 ng/mL) MDA-MB-231, but not in MDA-MB-468 cells on the transcription and protein levels. In conclusion, this study shows that fucoxanthin was more effective in MDA-MB-231 TNBC cells, where it can target VEGF-A and VEGF-C, inhibit cell proliferation and cell migration, and induce cell cycle arrest and apoptosis-the most crucial cellular processes involved in breast cancer development and progression.

The Anticancer Effects of Marine Carotenoid Fucoxanthin through Phosphatidylinositol 3-Kinase (PI3K)-AKT Signaling on Triple-Negative Breast Cancer Cells.

Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer that lacks specific targets such as estrogen, progesterone, and HER2 receptors. TNBC affects one in eight women in the United States, making up 15-20% of breast cancer cases. Patients with TNBC can develop resistance to chemotherapy over time, leading to treatment failure. Therefore, finding other options like natural products is necessary for treatment. The advantages of using natural products sourced from plants as anticancer agents are that they are less toxic, more affordable, and have fewer side effects. These products can modulate several cellular processes of the tumor microenvironment, such as proliferation, migration, angiogenesis, cell cycle arrest, and apoptosis. The phosphatidyl inositol 3-kinase (PI3K)-AKT signaling pathway is an important pathway that contributes to the survival and growth of the tumor microenvironment and is associated with these cellular processes. This current study examined the anticancer effects of fucoxanthin, a marine carotenoid isolated from brown seaweed, in the MDA-MB-231 and MDA-MB-468 TNBC cell lines. The methods used in this study include a cytotoxic assay, PI3K-AKT signaling pathway PCR arrays, and Wes analysis. Fucoxanthin (6.25 µM) + TNF-α (50 ng/mL) and TNF-α (50 ng/mL) showed no significant effect on cell viability compared to the control in both MDA-MB-231 and MDA-MB-468 cells after a 24 h treatment period. PI3K-AKT signaling pathway PCR array studies showed that in TNF-α-stimulated (50 ng/mL) MDA-MB-231 and MDA-MB-468 cells, fucoxanthin (6.25 µM) modulated the mRNA expression of 12 genes, including FOXO1, RASA1, HRAS, MAPK3, PDK2, IRS1, EIF4EBP1, EIF4B, PTK2, TIRAP, RHOA, and ELK1. Additionally, fucoxanthin significantly downregulated the protein expression of IRS1, EIF4B, and ELK1 in MDA-MB-231 cells, and no change in the protein expression of EIF4B and ELK1 was shown in MDA-MB-468 cells. Fucoxanthin upregulated the protein expression of RHOA in both cell lines. The modulation of the expression of genes and proteins of the PI3K-AKT signaling pathway may elucidate fucoxanthin's effects in cell cycle progression, apoptotic processes, migration, and proliferation, which shows that PI3K-AKT may be the possible molecular mechanism for fucoxanthin's effects. In conclusion, the results obtained in this study elucidate fucoxanthin's molecular mechanisms and indicate that fucoxanthin may be considered a promising candidate for breast cancer-targeted therapy.

Anticancer Effects of Fucoxanthin through Cell Cycle Arrest, Apoptosis Induction, Angiogenesis Inhibition, and Autophagy Modulation.

Cancer accounts for one in seven deaths worldwide and is the second leading cause of death in the United States, after heart disease. One of the standard cancer treatments is chemotherapy which sometimes can lead to chemoresistance and treatment failure. Therefore, there is a great need for novel therapeutic approaches to treat these patients. Novel natural products have exhibited anticancer effects that may be beneficial in treating many kinds of cancer, having fewer side effects, low toxicity, and affordability. Numerous marine natural compounds have been found to inhibit molecular events and signaling pathways associated with various stages of cancer development. Fucoxanthin is a well-known marine carotenoid of the xanthophyll family with bioactive compounds. It is profusely found in brown seaweeds, providing more than 10% of the total creation of natural carotenoids. Fucoxanthin is found in edible brown seaweed macroalgae such as Undaria pinnatifida, Laminaria japonica, and Eisenia bicyclis. Many of fucoxanthin's pharmacological properties include antioxidant, anti-tumor, anti-inflammatory, antiobesity, anticancer, and antihypertensive effects. Fucoxanthin inhibits many cancer cell lines' proliferation, angiogenesis, migration, invasion, and metastasis. In addition, it modulates miRNA and induces cell cycle growth arrest, apoptosis, and autophagy. Moreover, the literature shows fucoxanthin's ability to inhibit cytokines and growth factors such as TNF-α and VEGF, which stimulates the activation of downstream signaling pathways such as PI3K/Akt autophagy, and pathways of apoptosis. This review highlights the different critical mechanisms by which fucoxanthin inhibits diverse cancer types, such as breast, prostate, gastric, lung, and bladder development and progression. Moreover, this article reviews the existing literature and provides critical supportive evidence for fucoxanthin's possible therapeutic use in cancer.

The Role of Apoptotic Genes and Protein-Protein Interactions in Triple-negative Breast Cancer.

BACKGROUND/AIM: Compared to other breast cancer types, triple-negative breast cancer (TNBC) has historically had few treatment alternatives. Therefore, exploring and pinpointing potentially implicated genes could be used for treating and managing TNBC. By doing this, we will provide essential data to comprehend how the genes are involved in the apoptotic pathways of the cancer cells to identify potential therapeutic targets. Analysis of a single genetic alteration may not reveal the pathogenicity driving TNBC due to the high genomic complexity and heterogeneity of TNBC. Therefore, searching through a large variety of gene interactions enabled the identification of molecular therapeutic genes. MATERIALS AND METHODS: This study used integrated bioinformatics methods such as UALCAN, TNM plotter, PANTHER, GO-KEEG and PPIs to assess the gene expression, protein-protein interaction (PPI), and transcription factor interaction of apoptosis-regulated genes. RESULTS: Compared to normal breast tissue, gene expressions of BNIP3, TNFRSF10B, MCL1, and CASP4 were downregulated in UALCAN. At the same time, BIK, AKT1, BAD, FADD, DIABLO, and CASP9 was down-regulated in bc-GeneExMiner v4.5 mRNA expression (BCGM) databases. Based on GO term enrichment analysis, the cellular process (GO:0009987), which has about 21 apoptosis-regulated genes, is the top category in the biological processes (BP), followed by biological regulation (GO:0065007). We identified 29 differentially regulated pathways, including the p53 pathway, angiogenesis, apoptosis signaling pathway, and the Alzheimer's disease presenilin pathway. We examined the PPIs between the genes that regulate apoptosis; CASP3 and CASP9 interact with FADD, MCL1, TNF, TNFRSRF10A, and TNFRSF10; additionally, CASP3 significantly forms PPIs with CASP9, DFFA, and TP53, and CASP9 with DIABLO. In the top 10 transcription factors, the androgen receptor (AR) interacts with five apoptosis-regulated genes (p<0.0001; q<0.01), followed by retinoic acid receptor alpha (RARA) (p<0.0001; q<0.01) and ring finger protein (RNF2) (p<0.0001; q<0.01). Overall, the gene expression profile, PPIs, and the apoptosis-TF interaction findings suggest that the 27 apoptosis-regulated genes might be used as promising targets in treating and managing TNBC. Furthermore, from a total of 27 key genes, CASP2, CASP3, DAPK1, TNF, TRAF2, and TRAF3 were significantly correlated with poor overall survival in TNBC (p-value <0.05); they could play important roles in the progression of TNBC and provide attractive therapeutic targets that may offer new candidate molecules for targeted therapy. CONCLUSION: Our findings demonstrate that CASP2, CASP3, DAPK1, TNF, TRAF2, and TRAF3 were substantially associated with the overall survival rate (OS) difference of TNBC patients out of a total of 27 specific genes used in this study, which may play crucial roles in the development of TNBC and offer promising therapeutic interventions.

South African physiotherapists' perspectives on the competencies needed to work in special schools for learners with special needs.

BACKGROUND: Investigation into, and description of competencies in the various sectors in which the physiotherapy profession is practised, contribute to the standardisation of practice, professional education, and guides research and administration, and is necessary in South Africa. OBJECTIVE: To identify the competencies implemented by physiotherapists working in an educational setting for learners with special needs and to determine physiotherapists' opinions on the identified competencies. METHODS: A sequential mixed method research design was implemented to explore the competencies that physiotherapists implement during their intervention for children with special needs through focus group discussions (FGDs). A questionnaire based on the statements that emerged from the thematic analysis of the transcribed FGDs, and validated, was implemented in a cross-sectional survey amongst all physiotherapists employed in special schools. SPSS version 24 was used for the analysis of closed responses and thematic analysis was done on open-ended responses (n = 22). RESULTS: The respondents' knowledge and skills regarding physiotherapy theories and implementation ranged from 'good' to 'very good'. However, integration of the therapeutic knowledge and skills in different aspects of the special educational environment, and community integration, were rated 'poor' to 'fair'. Support of physiotherapists to implement policies and procedures, and to attend continuing professional development, ranged from 'fair' to 'poor'. CONCLUSION: Lack of knowledge in educational policies and procedures in classroom strategies negatively influence the integration of therapeutic strategies in the special educational environment. CLINICAL IMPLICATIONS: The contribution of our study to learners with special needs in schools was outlined.

Differential bacterial dynamics promote emergent community robustness to lake mixing: an epilimnion to hypolimnion transplant experiment.

Lake mixing disrupts chemical and physical gradients that structure bacterial communities. A transplant experiment was designed to investigate the influence of post-mixing environmental conditions and biotic interactions on bacterial community composition. The experimental design was 3x2 factorial, where water was incubated from three different sources (epilimnion, hypolimnion, and mixed epilimnion and hypolimnion) at two different locations in the water column (epilimnion or hypolimnion). Three replicate mesocosms of each treatment were removed every day for 5 days for bacterial community profiling, assessed by automated ribosomal intergenic spacer analysis. There were significant treatment effects observed, and temperature was the strongest measured driver of community change (r=-0.66). Epilimnion-incubated communities changed more than hypolimnion-incubated. Across all treatments, we classified generalist, layer-preferential and layer-specialist populations based on occurrence patterns. Most classified populations were generalists that occurred in both strata, suggesting that communities were robust to mixing. In a network analysis of the mixed-inocula treatments, there was correlative evidence of inter-population biotic interactions, where many of these interactions involved generalists. These results reveal differential responses of bacterial populations to lake mixing and highlight the role of generalist taxa in structuring an emergent community-level response.

Provision of relapse prevention interventions in UK NHS Stop Smoking Services: a survey.

BACKGROUND: UK NHS Stop Smoking Services provide cost effective smoking cessation interventions but, as yet, there has been no assessment of their provision of relapse prevention interventions. METHODS: Electronic questionnaire survey of 185 UK Stop Smoking Services Managers. RESULTS: Ninety six Stop Smoking Service managers returned completed questionnaires (52% response rate). Of these, 58.3% (n = 56) ran NHS Stop Smoking Services which provided relapse prevention interventions for clients with the most commonly provided interventions being behavioural support: telephone (77%), group (73%), and individual (54%). Just under half (48%, n = 27) offered nicotine replacement therapy (NRT), 21.4% (n = 12) bupropion; 19.6% (n = 11) varenicline. Over 80% of those providing relapse prevention interventions do so for over six months. Nearly two thirds of all respondents thought it was likely that they would either continue to provide or commence provision of relapse prevention interventions in their services. Of the remaining respondents, 66.7% (n = 22) believed that the government focus on four-week quit rates, and 42.9% (14 services) believed that inadequate funding for provision of relapse prevention interventions, were major barriers to introducing these interventions into routine care. CONCLUSIONS: Just over half of UK managers of NHS Stop Smoking Services who responded to the questionnaire reported that, in their services, relapse prevention interventions were currently provided for clients, despite, at that time, there being a weak evidence base for their effectiveness. The most commonly provided relapse prevention interventions were those for which there was least evidence. If these interventions are found to be effective, barriers would need to be removed before they would become part of routine care.

Relapse prevention in UK Stop Smoking Services: a qualitative study of health professionals' views and beliefs.

BACKGROUND: NHS Stop Smoking Services in the UK provide cost effective smoking cessation interventions, but approximately 75% of smokers who are abstinent at 4 weeks relapse to smoking by 12 months. This study aimed to explore health professionals' understanding of relapse prevention interventions (RPIs), the feasibility of offering such support and whether and how these are currently used in UK NHS Stop Smoking Services. METHODS: Sixteen health professionals working in UK NHS Stop Smoking Services, selected from those attending a national conference were interviewed and Framework Analysis was used to identify recurrent key themes and concepts in their perceptions and experiences of providing relapse prevention interventions (RPIs). RESULTS: Interviewees had diverse perceptions of relapse prevention as a concept. Whilst relapse prevention was largely seen as support to prevent abstinent smokers from relapsing to smoking, some interviewees stated that RPIs were being delivered to lapsed or relapsed smokers. Current provision of RPIs was most commonly described as behavioural counselling being offered predominantly after completed cessation treatment, often in the format of 'rolling groups' which the client was encouraged to attend. Commonly identified barriers to the introduction of RPIs were funding and government targets focussed on short term cessation, smokers' low uptake of offered RPIs and an uncertain evidence base for their effectiveness. Interviewees were positive about the potential use of pharmacotherapy for relapse prevention, but were negative about the possibility of introducing proactive telephone counselling for this purpose. CONCLUSION: There is currently no shared understanding of the concept of relapse prevention amongst this sample of health professionals working in UK NHS Stop Smoking Services. For RPIs to be systematically delivered via these services, a commonly-held, widely-accepted and understood definition of relapse prevention would be needed. Other barriers towards introducing RPIs, such as their weak evidence and the short term cessation-focussed targets against which UK stop smoking services are measured, would also need addressing and interventions which are acceptable to abstinent smokers would need to be developed.

Community structure explains antibiotic resistance gene dynamics over a temperature gradient in soil.

Soils are reservoirs of antibiotic resistance genes (ARGs), but environmental dynamics of ARGs are largely unknown. Long-term disturbances offer opportunities to examine microbiome responses at scales relevant for both ecological and evolutionary processes and can be insightful for studying ARGs. We examined ARGs in soils overlying the underground coal seam fire in Centralia, PA, which has been burning since 1962. As the fire progresses, previously hot soils can recover to ambient temperatures, which creates a gradient of fire impact. We examined metagenomes from surface soils along this gradient to examine ARGs using a gene-targeted assembler. We targeted 35 clinically relevant ARGs and two horizontal gene transfer-related genes (intI and repA). We detected 17 ARGs in Centralia: AAC6-Ia, adeB, bla_A, bla_B, bla_C, cmlA, dfra12, intI, sul2, tetA, tetW, tetX, tolC, vanA, vanH, vanX and vanZ. The diversity and abundance of bla_A, bla_B, dfra12 and tolC decreased with soil temperature, and changes in ARGs were largely explained by changes in community structure. We observed sequence-specific biogeography along the temperature gradient and observed compositional shifts in bla_A, dfra12 and intI. These results suggest that increased temperatures can reduce soil ARGs but that this is largely due to a concomitant reduction in community-level diversity.

Abstinence and relapse among smokers who use varenicline in a quit attempt-a pooled analysis of randomized controlled trials.

BACKGROUND AND AIMS: Varenicline increases the likelihood of long-term abstinence following a quit attempt. It has been suggested that (1) part of its benefit arises from 'recruiting into abstinence' smokers who are not able to stop on the target quit date and (2) there may be a higher rate of relapse after treatment. This study addressed these issues. METHODS: Meta-analyses of data from randomized controlled trials (RCTs) of varenicline identified from the 2012 Cochrane review of nicotine receptor partial agonists for smoking cessation were used to compare the abstinence and relapse patterns on active drug and placebo. Studies of varenicline compared with placebo in adult daily smokers with longest follow-up at either 6 or 12 months and with at least three follow-ups in the first month were included. Biochemically verified abstinence rates at each of six follow-up time-points (2, 3, 4, 12, 24 and 52 weeks) were pooled for studies reporting point prevalence abstinence. Biochemically verified abstinence rates at three follow-up time-periods (9-12 weeks, 9-24 weeks and 9-52 weeks) were pooled for studies reporting continuous abstinence. Random effects meta-analysis was used to estimate pooled proportions with 95% confidence intervals. RESULTS: Nineteen RCTs were included. In varenicline-treated participants, point-prevalence abstinence increased by 22 percentage points from week 2 [32%: 95% confidence interval (CI) = 25-40%] to week 12 (54%: 95% CI = 48-61%). The increase was 8 percentage points in the placebo group: 16% (95% CI = 11-21%) to 24% (95% CI = 17-33%). In varenicline-treated participants the relapse from weeks 9-12 to week 52 was 55%: 49% abstinent in weeks 9-12 (95% CI = 45-53%) versus 22% at week 52 (95% CI = 19-25%). In placebo-treated participants it was 53%: 17% (95% CI = 13-25%) in weeks 9-12 versus 8% (95% CI = 6-12%) at week 52. CONCLUSIONS: Varenicline recruits smokers into abstinence following the target quit date to a greater extent than placebo. Relapse rates from end of treatment to 52 weeks are similar in varenicline- and placebo-treated smokers.

The fractional postcoital test: a reappraisal.

Postcoital tests (PCTs) were conducted following artificial insemination with either live or dead spermatozoa. The cervical mucus samples were analyzed for sperm counts and acid phosphatase activity. The results indicate that the final distribution of mucus and sperm in the PCT sampling catheter is not truly representative of their original distribution within the cervix. Moreover, PCTs done after insemination with semen containing only dead sperm yield unrealistically high sperm counts and high cervical mucus acid phosphatase levels. This is apparently due to the translocation of semen from the area of the external os into the mucus sample by the sampling catheter. This problem can be largely prevented by removing a plug of mucus from the external os prior to the catheter sampling procedure.

Papillary serous carcinoma of the cervix: ultrasonographic findings.

BACKGROUND: Papillary serous carcinoma of the uterine cervix is a rare malignancy only recently described. Optimal treatment is unknown. CASE REPORT: A 65 year old white female, approximately 15 years postmenopausal, presented with a recent history of postmenopausal bleeding. Cervical cytology was negative for any abnormalities. Ultrasonography of the pelvis revealed a thickened anterior endometrium with fluid present along with a papillary mass in the endocervix. Hysteroscopy, dilatation and curettage revealed a low uterine segment mass consistent with a pathologic diagnosis of grade 2 endometrioid adenocarcinoma of the endometrium. A total abdominal hysterectomy, bilateral salpingo-oophorectomy and bilateral pelvic and paraaortic lymphadenectomy revealed a papillary serous carcinoma of the cervix metastatic to pelvic and paraaortic lymph nodes. CONCLUSION: Cervical papillary serous carcinoma can be discovered pre-operatively by ultrasonography. Because of its relatively recent description and relative rarity, optimal treatment is unknown.

Gynecologic and obstetric problems of the female dancer.

The effect of athletics and exercise on women is a new area of medicine, and little factual information is currently available. This discussion briefly reviews some of the gynecologic and obstetric information available to give guidelines for the woman involved in strenuous physical activity.

PIP: This paper reviews the gynecologic and obstetric literature on problems encountered by women involved in strenuous physical exercise. Those who begin training prior to puberty tend to have delayed menarche as compared with nonactive controls and to experience significantly more menstrual dysfunction. Attainment of a minimum weight or a certain percentage of body fat appears to be an important factor. Increased androgen levels resulting from stress on the adrenal gland or ovary may affect the female athlete's hypothalamus, and weight loss may cause additional abnormalities of the endocrine system. Physical examination of athletes with menstrual dysfunction should include evaluation of height and weight, general nutrition, the presence or absence of galactorrhea, and the presence of obvious hormonal abnormalities. Management of the pregnant athlete is controversial; however, there is general agreement that a woman can continue athletic activity at medium effort until the 8th month of pregnancy, at which point activity should be reduced until the 4th week postpartum. Barrier contraception is generally recommended for physically active women. Currently, there is little factual information on the effects of athletics and exercise on women. For example, it is not known whether there is a lasting negative effect on the young dancer's menstrual cycles and her reproductive capacity after discontinuation of active training. Until more research evidence is accumulated, careful medical follow-up of female dancers can enable them to safely perform with minimal risk of present or future health problems.

The effect of simplified adhesives on the bond strength to dentin of dual-cure resin cements.

The purpose of this study was to compare the shear bond strengths to dentin of two dual-cure resin cements, one with a unique initiator, NX3 (Kerr Corp), and the other with a traditional redox-initiator system, Calibra (Dentsply), when used in combination with simplified or nonsimplified adhesive agents. The two dual-cure resin cements, in either self- or dual-cure activation modes, were bonded to human dentin with four dental adhesives to create 16 subgroups of 10 specimens each. After 24 hours of storage in distilled water at 37°C, the specimens were tested in shear in a universal testing machine. With both NX3 and Calibra, bond strengths significantly increased when the specimens were dual cured. In addition, with either cement in either mode, the nonsimplified adhesives performed significantly better than did the simplified adhesive bonding agents. When used specifically with simplified adhesives in either cure mode, NX3 did not produce significantly higher bond strengths than did Calibra. In general, lower dentin bond strengths were found with simplified adhesives or self-cure activation with either resin cement.