title: Gene expression profiles of 3D gene-edited bioprinted cells

authors: Turnbull, Catherine

subjects: Bioprinting

abstract: Cancer is the biggest cause of death in New Zealand each year and places a large economic strain on the health system. Previously, two-dimensional (2D) cell models have provided insight into the development of cancer but now, three-dimensional (3D) models provided more accurate information on how cell-cell interactions in the body affect the efficacy of different drug therapies. This research aims to investigate the gene expression profiles of 3D gene-edited bioprinted lung cancer cells to understand how mutations and the vital trace element Selenium (Se) affects gene expression. The human A549 lung cancer cell line was bioprinted into a set dome shape made of sodium alginate, gelatine, and cell media. The cells were then cultured for 21 days before a six-hour drug exposure with 2μM and 10μM Methylselenic Acid (MSA).
RNA was also extracted from 18 bioprints and the spectrophotometer data suggested high quality with a total RNA concentration range of 9 – 671μg/μL, respectively. The RNA integrity was further examined using the RNA tape station. The solvent control (n=2) and the 2μM MSA treatment (n=3) from the second independent experiment resulted in intact ribosomal bands. The protein concentration was determined using a Bradford assay and ranged from 0.1 – 3.42mg/mL. The electrophoresed protein showed a molecular weight size range from 120kDa – 10kDa. The protein was then transferred onto a western blot membrane and detected by two primary antibodies; (1) HSP70 - correctly fold proteins that are under cellular stress and (2) GAPDH – a loading control reference antibody. The HSP70 protein expression was measured for the drug-treated bioprints for three bioprinting experiment. We hypothesised an upregulation in HSP70 expression based on previous studies. However, the data set could not be replicated to conclusively downregulation in response to 2 and 10μM MSA. Thus, further optimisation into drug accessibility, cell and protein loading concentration is required.
Gene-editing using lipofectamine and CRISPR-Cas9 was attempted using A549 cells but no conclusive evidence that a mutation had been introduced into the Cyclin Dependent Kinase 4 (CDK4) gene was observed (n=5). Three research optimisations have been recommended for gene-editing and establishing a stable gene-edited cell line for future use in 3D bioprinting and gene expression analysis.

text: Gene expression profiles of 3D gene-edited bioprinted cells Cancer is the biggest cause of death in New Zealand each year and places a large economic strain on the health system. Previously, two-dimensional (2D) cell models have provided insight into the development of cancer but now, three-dimensional (3D) models provided more accurate information on how cell-cell interactions in the body affect the efficacy of different drug therapies. This research aims to investigate the gene expression profiles of 3D gene-edited bioprinted lung cancer cells to understand how mutations and the vital trace element Selenium (Se) affects gene expression. The human A549 lung cancer cell line was bioprinted into a set dome shape made of sodium alginate, gelatine, and cell media. The cells were then cultured for 21 days before a six-hour drug exposure with 2μM and 10μM Methylselenic Acid (MSA).
RNA was also extracted from 18 bioprints and the spectrophotometer data suggested high quality with a total RNA concentration range of 9 – 671μg/μL, respectively. The RNA integrity was further examined using the RNA tape station. The solvent control (n=2) and the 2μM MSA treatment (n=3) from the second independent experiment resulted in intact ribosomal bands. The protein concentration was determined using a Bradford assay and ranged from 0.1 – 3.42mg/mL. The electrophoresed protein showed a molecular weight size range from 120kDa – 10kDa. The protein was then transferred onto a western blot membrane and detected by two primary antibodies; (1) HSP70 - correctly fold proteins that are under cellular stress and (2) GAPDH – a loading control reference antibody. The HSP70 protein expression was measured for the drug-treated bioprints for three bioprinting experiment. We hypothesised an upregulation in HSP70 expression based on previous studies. However, the data set could not be replicated to conclusively downregulation in response to 2 and 10μM MSA. Thus, further optimisation into drug accessibility, cell and protein loading concentration is required.
Gene-editing using lipofectamine and CRISPR-Cas9 was attempted using A549 cells but no conclusive evidence that a mutation had been introduced into the Cyclin Dependent Kinase 4 (CDK4) gene was observed (n=5). Three research optimisations have been recommended for gene-editing and establishing a stable gene-edited cell line for future use in 3D bioprinting and gene expression analysis.

year: 2023

title: Sexual Desire in Women Presenting for Antiandrogen Therapy

authors: Conaglen, Helen M.

subjects: antiandrogen therapy

abstract: Women presenting with hirsuties/polycystic ovary syndrome have increased production of androgens. Clinical lore suggests that these women may have increased sexual desire. Treatment of hirsuties commonly involves antiandrogen therapy, a form of therapy with a potential for reducing sexual desire. The present study investigated sexual desire in 29 hirsute women aged 19 to 43 years presenting for therapy. We conducted a questionnaire appraisal of the women’s sexual desire, body and self-esteem, and affect at baseline, 3 months, and 12 months and compared the data with a control group of 30 nonhirsute women of similar mean age. Those in the treatment group also had their Ferriman and Gallwey scores and body mass indices calculated at baseline and end of study for those in the treatment group. We determined hormone levels for those in the treatment group with baseline blood tests. Our hypotheses were that the hirsute women would experience different levels of sexual desire than the control group prior to therapy and that therapy would have a demonstrable effect on the self-reported sexual desire of these women. The study demonstrated that women with hirsuties had mean levels of sexual desire and body esteem that were significantly lower than the control group women. During the year-long course of therapy, the sexual desire levels of the hirsute women decreased progressively, while their self-esteem increased. The women’s Ferriman and Gallwey scores fell, indicating diminishing hirsutism. These findings provide empirical data upon which clinicians can base advice to patients seeking therapy.

text: Sexual Desire in Women Presenting for Antiandrogen Therapy Women presenting with hirsuties/polycystic ovary syndrome have increased production of androgens. Clinical lore suggests that these women may have increased sexual desire. Treatment of hirsuties commonly involves antiandrogen therapy, a form of therapy with a potential for reducing sexual desire. The present study investigated sexual desire in 29 hirsute women aged 19 to 43 years presenting for therapy. We conducted a questionnaire appraisal of the women’s sexual desire, body and self-esteem, and affect at baseline, 3 months, and 12 months and compared the data with a control group of 30 nonhirsute women of similar mean age. Those in the treatment group also had their Ferriman and Gallwey scores and body mass indices calculated at baseline and end of study for those in the treatment group. We determined hormone levels for those in the treatment group with baseline blood tests. Our hypotheses were that the hirsute women would experience different levels of sexual desire than the control group prior to therapy and that therapy would have a demonstrable effect on the self-reported sexual desire of these women. The study demonstrated that women with hirsuties had mean levels of sexual desire and body esteem that were significantly lower than the control group women. During the year-long course of therapy, the sexual desire levels of the hirsute women decreased progressively, while their self-esteem increased. The women’s Ferriman and Gallwey scores fell, indicating diminishing hirsutism. These findings provide empirical data upon which clinicians can base advice to patients seeking therapy.

year: 2003

title: Putting it in there: "PSYC575 Indigenous Psychologies" as a for-credit university paper in New Zealand

authors: Waitoki, Waikaremoana

abstract: Over the past 30 years, scholar advocates have sparked the emergence of indigenous psychology as an important contribution to understanding the experiences and challenges faced by indigenous peoples. Part of this exercise is to also bring indigenous knowledges to the learning table as critical information for meaning making and resolving psychological challenges. Increasingly, academics are writing and teaching papers in the fledgling field as important and legitimate contributions for undergraduate and graduate degree credit. Previously, students and academics were confined to the more flexible research driven, directed-learning, study options rather than papers explicitly advertised and institutionally promoted. In this paper, we discuss how we are going about writing the curricula for a graduate level paper called "Indigenous Psychologies" to be offered in 2016 at the University of Waikato, New Zealand. We discuss a) principles that guide curricula decision-making; b) the programme of lecture topics developed; and c) finding and developing supporting resources. We welcome conversation and resource sharing.

text: Putting it in there: "PSYC575 Indigenous Psychologies" as a for-credit university paper in New Zealand Over the past 30 years, scholar advocates have sparked the emergence of indigenous psychology as an important contribution to understanding the experiences and challenges faced by indigenous peoples. Part of this exercise is to also bring indigenous knowledges to the learning table as critical information for meaning making and resolving psychological challenges. Increasingly, academics are writing and teaching papers in the fledgling field as important and legitimate contributions for undergraduate and graduate degree credit. Previously, students and academics were confined to the more flexible research driven, directed-learning, study options rather than papers explicitly advertised and institutionally promoted. In this paper, we discuss how we are going about writing the curricula for a graduate level paper called "Indigenous Psychologies" to be offered in 2016 at the University of Waikato, New Zealand. We discuss a) principles that guide curricula decision-making; b) the programme of lecture topics developed; and c) finding and developing supporting resources. We welcome conversation and resource sharing.

year: 2015