Potent cytotoxic effects of Calomeria amaranthoides on ovarian cancers

<p>Abstract</p> <p>Background</p> <p>Ovarian cancer remains the leading cause of death from gynaecological malignancy. More than 60% of the patients are presenting the disease in stage III or IV. In spite of combination of chemotherapy and surgery the prognosis stays poor for therapy regimen.</p> <p>Methods</p> <p>The leaves of a plant endemic to Australia, <it>Calomeria amaranthoides</it>, were extracted and then fractionated by column chromatography. <it>In vitro </it>cytotoxicity tests were performed with fractions of the plant extract and later with an isolated compound on ovarian cancer cell lines, as well as normal fibroblasts at concentrations of 1-100 μg/mL (crude extract) and 1-10 μg/mL (compound). Cytotoxicity was measured after 24, 48 and 72 hours by using a non-fluorescent substrate, Alamar blue.</p> <p><it>In vivo </it>cytotoxicity was tested on ascites, developed in the abdomen of nude mice after inoculation with human OVCAR₃cells intraperitoneally. The rate of change in abdomen size for the mice was determined by linear regression and statistically evaluated for significance by the unpaired t test.</p> <p>Results</p> <p>Two compounds were isolated by chromatographic fractionation and identified by ¹H-NMR, ¹³C-NMR and mass spectrometry analyses, EPD, an α-methylene sesquiterpene lactone of the eremophilanolide subtype, and EPA, an α-methylene carboxylic acid.</p> <p>Cytotoxicity of EPD for normal fibroblasts at all time points IC₅₀was greater than 10 μg/mL, whereas, for OVCAR₃cells at 48 hours IC₅₀was 5.3 μg/mL (95% confidence interval 4.3 to 6.5 μg/mL).</p> <p>Both, the crude plant extract as well as EPD killed the cancer cells at a final concentration of 10 μg/mL and 5 μg/mL respectively, while in normal cells only 20% cell killing effect was observed. EPA had no cytotoxic effects.</p> <p>Changes in abdomen size for control versus Cisplatin treated mice were significantly different, P = 0.023, as were control versus EPD treated mice, P = 0.025, whereas, EPD versus Cisplatin treated mice were not significantly different, P = 0.13.</p> <p>Conclusions</p> <p>For the first time both crude plant extract from <it>Calomeria amaranthoides </it>and EPD have been shown to have potent anti-cancer effects against ovarian cancer.</p

KIAA1109 Variants Are Associated with a Severe Disorder of Brain Development and Arthrogryposis.

Whole-exome and targeted sequencing of 13 individuals from 10 unrelated families with overlapping clinical manifestations identified loss-of-function and missense variants in KIAA1109 allowing delineation of an autosomal-recessive multi-system syndrome, which we suggest to name Alkuraya-Kučinskas syndrome (MIM 617822). Shared phenotypic features representing the cardinal characteristics of this syndrome combine brain atrophy with clubfoot and arthrogryposis. Affected individuals present with cerebral parenchymal underdevelopment, ranging from major cerebral parenchymal thinning with lissencephalic aspect to moderate parenchymal rarefaction, severe to mild ventriculomegaly, cerebellar hypoplasia with brainstem dysgenesis, and cardiac and ophthalmologic anomalies, such as microphthalmia and cataract. Severe loss-of-function cases were incompatible with life, whereas those individuals with milder missense variants presented with severe global developmental delay, syndactyly of 2nd and 3rd toes, and severe muscle hypotonia resulting in incapacity to stand without support. Consistent with a causative role for KIAA1109 loss-of-function/hypomorphic variants in this syndrome, knockdowns of the zebrafish orthologous gene resulted in embryos with hydrocephaly and abnormally curved notochords and overall body shape, whereas published knockouts of the fruit fly and mouse orthologous genes resulted in lethality or severe neurological defects reminiscent of the probands' features.This article is freely available via Open Access. Click on the Additional Link above to access the full-text via the publisher's site

Mullerin neoplasia of the ovaries

In order to develop models of human ovarian cancer, attempts were made to establish a variety of neoplastic ovarian tissues as cell lines and as xenografts in nude mice. The ig_yitrg and in_yiyg phenotype of these cells was then characterised using cytogenetics, and DNA flow cytometry. A number of differentiation and tumour progression-related cell surface and cytoplasmic markers were also monitored. Cell invasive properties were studied using" two recently developed 'techniques to investigate metastatic behaviour. Preliminary efforts were made to develop monoclonal antibodies to the JoN cell line. Long-term cell and xenograft lines were successfully established from a high proportion of ovarian carcinoma patients studied, whether derived from solid tumours or malignant effusions. All cell lines grew as monolayers with variable doubling time and morphology. There were seven serous carcinomas (JN, JC, JC P/E, NW, JV, JG and LMcM), two carcinosarcomas (JoN and NF) and one clear cell carcinoma (GG) established and characterised. In the case of the JoN cell line two subpopulations separated by density—gradient centrifugation were established as cell lines and xenografts. Primary cultures (up to three months) were also established from five borderline ovarian tumours and from normal fetal and post—menopausal ovary. In general all the lines and cultures were similar morphologically to their tumours of origin, however the carcinosarcoma-derived lines showed predominantly epithelial features. Regardless of the DNA content of the original tumour, as monitored by flow cytometry, none of the lines established in culture remained diploid beyond the first 3-4 passages. The commonest cytogenetic abnormalities among the cultured cell lines were deletions of chromosome 1, followed by chromosomes 3, 4 and 7. Double minutes were frequent and a 13p duplication developed during culture of one line (JON). There was no single cytogenetic abnormality common to all lines. Epithelial membrane antigen (EMA) and keratin were commonly expressed among the established cell lines, especially in those from serous borderline tumours. Vimentin was detected in six lines, usually in conjunction with keratin. In contrast to the serous carcinoma lines, one subline of the JoN carcinosarcoma line (HDF) stained positive with desmin

New insights in phenotype and treatment of lung disease immuno-deficiency and chromosome breakage syndrome (LICS)

We report five patients with lung disease immuno-deficiency and chromosome breakage syndrome (LICS) but without recurrent infections and severe immunodeficiency. One patient had extended survival to 6.5 years. Hematopoietic stem-cell transplantation failed to cure another patient. Our findings suggest that the immunological abnormalities can be limited and do not fully explain the LICS phenotype

Loss of syntaxin 3 causes variant microvillus inclusion disease

Microvillus inclusion disease (MVID) is a disorder of intestinal epithelial differentiation characterized by life-threatening intractable diarrhea. MVID can be diagnosed based on loss of microvilli, microvillus inclusions, and accumulation of subapical vesicles. Most patients with MVID have mutations in myosin Vb that cause defects in recycling of apical vesicles. Whole-exome sequencing of DNA from patients with variant MVID showed homozygous truncating mutations in syntaxin 3 (STX3). STX3 is an apical receptor involved in membrane fusion of apical vesicles in enterocytes. Patient-derived organoid cultures and overexpression of truncated STX3 in Caco-2 cells recapitulated most characteristics of variant MVID. We conclude that loss of STX3 function causes variant MVID

Cost-effectiveness of laparoscopy as diagnostic tool before primary cytoreductive surgery in ovarian cancer

Objective. To evaluate the cost-effectiveness of a diagnostic laparoscopy prior to primary cytoreductive surgery to prevent futile primary cytoreductive surgery (i.e. leaving >1 cm residual disease) in patients suspected of advanced stage ovarian 'cancer. Methods. An economic analysis was conducted alongside a randomized controlled trial in which patients suspected of advanced stage ovarian cancer who qualified for primary cytoreductive surgery were randomized to either laparoscopy or primary cytoreductive surgery. Direct medical costs from a health care perspective over a 6-month time horizon were analyzed. Health outcomes were expressed in quality-adjusted life-years (QALYs) and utility was based on patient's response to the EQ-5D questionnaires. We primarily focused on direct medical costs based on Dutch standard prices. Results. We studied 201 patients, of whom 102 were randomized to laparoscopy and 99 to primary cytoreductive surgery. No significant difference in QALYs (utility = 0.01; 95% CI 0.006 to 0.02) was observed. Laparoscopy reduced the number of futile laparotomies from 39% to 10%, while its costs were (sic) 1400 per intervention, making the overall costs of both strategies comparable (difference (sic) - 80 per patient (95% CI - 470 to 300)). Findings were consistent across various sensitivity analyses. Conclusion. In patients with suspected advanced stage ovarian cancer, a diagnostic laparoscopy reduced the number of futile laparotomies, without increasing total direct medical health care costs, or adversely affecting complications or quality of life

Laparoscopy to Predict the Result of Primary Cytoreductive Surgery in Patients With Advanced Ovarian Cancer:A Randomized Controlled Trial

Purpose To investigate whether initial diagnostic laparoscopy can prevent futile primary cytoreductive surgery (PCS) by identifying patients with advanced-stage ovarian cancer in whom . 1 cm of residual disease will be left after PCS. Patients and Methods This multicenter, randomized controlled trial was undertaken within eight gynecologic cancer centers in the Netherlands. Patients with suspected awdvanced-stage ovarian cancer who qualified for PCS were eligible. Participating patients were randomly assigned to either laparoscopy or PCS. Laparoscopy was used to guide selection of primary treatment: Either primary surgery or neoadjuvant chemotherapy followed by interval surgery. The primary outcome was futile laparotomy, defined as a PCS with residual disease of.1 cm. Primary analyses were performed according to the intention-To-Treat principle. Results Between May 2011 and February 2015, 201 participants were included, of whom 102 were assigned to diagnostic laparoscopy and 99 to primary surgery. In the laparoscopy group, 63 (62%) of 102 patients underwent PCS versus 93 (94%) of 99 patients in the primary surgery group. Futile laparotomy occurred in 10 (10%) of 102 patients in the laparoscopy group versus 39 (39%) of 99 patients in the primary surgery group (relative risk, 0.25; 95% CI, 0.13 to 0.47; P , .001). In the laparoscopy group, three (3%) of 102 patients underwent both primary and interval surgery compared with 28 (28%) of 99 patients in the primary surgery group (P , .001). Conclusion Diagnostic laparoscopy reduced the number of futile laparotomies in patients with suspected advanced-stage ovarian cancer. In women with a plan for PCS, these data suggest that performance of diagnostic laparoscopy first is reasonable and that if cytoreduction to , 1 cm of residual disease seems feasible, to proceed with PCS