Novel biomarkers in two gastrointestinal malignancies - the potential of early detection and identification of new therapies

Currently, there are no sufficient biomarkers that can be utilised in hepatocellular cancer (HCC) and neuroendocrine tumours (NETs) that can help in making an early diagnosis, or recognise serious complications that can be associated with the tumour. HCC is usually diagnosed at an advanced stage when curative treatment options are limited. There is no recommended biomarker to use in clinical practice to diagnose early HCC at present. NETs can occur in different sites, but the gastroenteropancreatic NETs are the commonest sites of origin. Serious complications can be associated with this cancer due to the underlying gastrointestinal and cardiac valve fibrosis, including intestinal obstruction and ischaemia, and carcinoid heart disease (CHD). Within this thesis the suitability of prothrombin induced by vitamin K absence-II (PIVKA-II) as a biomarker in HCC will be presented. The aims of this study are to compare PIVKA-II to the traditional marker alpha-fetoprotein (AFP) in diagnosing early HCC, as well as a combination of both biomarkers, assess PIVKA-II levels in patients undergoing ablation therapy, and assess if levels of PIVKA-II increase with disease progression of HCC. The second project in this thesis describes the suitability of vascular adhesion-protein 1 (VAP-1) as a biomarker in NETs and CHD. The aims of this project are to compare the levels of soluble VAP-1 (sVAP-1) in different NET cohorts and healthy volunteers and compare levels of sVAP-1 pre- and post-treatment in midgut NETs. I also aim to study the tissue expression of VAP-1 in midgut NET tissue and CHD. AFP has been the traditional marker used in HCC surveillance. PIVKA-II was not found to be a superior marker to AFP in diagnosing early HCC, and the combination of both markers was only slightly better compared to AFP alone. However, on explant histology, pre-transplant serum PIVKA-II levels were found to be significantly higher in the presence of microvascular invasion and also in moderately- or poorly- differentiated HCC. This was not seen with AFP. In the longitudinal data analysis, PIVKA-II shows a promising role in the surveillance of recurrent HCC post-ablation. Tissue expression of VAP-1 was present in midgut NETs and in the CHD valves, associated with the presence of a dense stromal and collagen network. Significantly higher % area of tissue expression of VAP-1 was seen in CHD valves compared to control valves. Across the healthy controls, midgut NETs and CHD groups, significantly higher levels of circulating sVAP-1 was seen with highest levels found in CHD. The longitudinal data analysis did not show any significant change in the sVAP-1 levels between the pre- and post-treatment samples, similar to the markers currently used in clinical practice (chromogranin A, 24 hour urinary 5-hydroxyindole acetic acid and N-terminal pro b-type natriuretic peptide). PIVKA-II could therefore be a prognostic marker in HCC and may support patient stratification for therapy. Despite a small number in the longitudinal data analysis, PIVKA-II shows a promising role in the surveillance of recurrent HCC post-ablation therapy. The VAP-1 study identifies the presence of VAP-1, both in a circulating form and tissue expression in midgut NETs and in CHD, and therefore could be a potential biomarker and treatment target in NETs

STUDIES ON ENDOPHYTES AND ANTIBACTERIAL ACTIVITY OF SAUSSUREA COSTUS (FALC.)

Different plant parts (root, stem, leaf) of Saussurea costus were used to isolate and investigate endophytic fungal species in summer, rainy and winter seasons. Total ten species of endophytic fungi belonging to seven genera (Aspergillus, Cunninghamella, Myrothecium, Penicillium, Pythium, Rhizopus and Trichoderma) were isolated from the root, stem and leaf in different seasons. The genus Aspergillus was found to be dominant with three species (A. nustus, A. wentii and A. niger). The genus Rhizopus was represented by two species (R. oryzae, R. nigricans).The genera Cunninghamella, Myrothecium, Penicillium, Trichoderma and Pythium were represented by one species each i.e. Cunninghamella elegans, Myrothecium roridum, Penicillium chrysogenum, Trichoderma viride and Pythium sp. respectively. Antibacterial activity of root of Saussurea costus has been investigated using different solvents (methanol, ethanol and acetone) at different concentrations (25%, 50%, 75% and 100%) against three test bacteria namely S. aureus, E. coli and Y. pestis. Root extract of S. costus showed greater antibacterial activity using methanol as solvent followed by acetone and ethanol solvent. In case of methanol extract, maximum inhibition activity was shown against S. aureus and minimum against E. coli. Maximum inhibition was shown against Y. pestis and minimum against E. coli in case of acetone extract whereas ethanol extract exhibited highest inhibitory activity against E. coli and lowest against Y. pestis. On the basis of results compiled above, it is evident that S. costus exhibited antibacterial activity against all the test bacteria in all the solvents used in this study thereby conforming it a good antibacterial agent for future study.   KEY WORDS: Saussurea costus, endophyte isolation, a

Review of randomized controlled trials on psychological interventions in child sexual abuse: Current status and emerging needs in the Indian context

Child sexual abuse (CSA) is a critical, psychologically traumatic and sometimes life-threatening incident often associated with sequel of adverse physical, behavioral, and mental health consequences. Factors such as developmental age of the child, severity of abuse, closeness to the perpetrator, availability of medico-legal-social support network and family care, gender stereotypes in the community complicate the psychological trauma. Although the research on the effects of CSA as well as psychological intervention to reduce the victimization and promote the mental health of the child is in its infancy stage in India, the global research in the past three decades has progressed much ahead. A search was performed using MEDLINE, PubMed, PsycINFO, and Google Scholar from 1984 to 2015 and only 17 randomized controlled trials (RCTs) out of 96 potentially relevant studies were included. While nonspecific therapies covering a wide variety of outcome variables were prominent till 1999s, the trend changed to specific and focused forms of trauma-focused therapies in next one-and-half decades. Novel approaches to psychological interventions have also been witnessed. One intervention (non-RCT) study on effects on general counseling has been reported from India

Dentin post: A new method for reinforcing the tooth

Ideal coronal reconstruction of a fractured tooth is still a challenge for restorative dentistry. Many post systems are available but none of them have all the ideal biological and mechanical properties. In this case report a "dentin post" was used for the reinforcement of fractured endodontically treated tooth by virtue of its biomimetic property. Following root canal treatment, a post space was prepared. A direct wax impression was made of the post space. A freshly extracted maxillary cuspid was used to prepare a dentin post, which was then verified in the post space. The dentin post was then luted with dual-cure resin in the canal. Biological dentin posts are considered a good alternative to conventional post systems as they preserve internal dentin walls and provide excellent adhesion and resilience, similar to the natural tooth structure

Development of SMEDDS using natural lipophile: application to β-Artemether delivery

The objective of the present investigation was to formulate self-microemulsifying drug delivery systems (SMEDDS) using a novel, indigenous natural lipophile (N-LCT) as an oily phase. SMEDDS based on N-LCT and commercially available modified oil (Capryol 90) were formulated and their application in improving the delivery of a lipophilic anti-malarial drug, β-Artemether (BAM) was also evaluated. BAM-loaded SMEDDS were characterized with respect to mean globule size and in vitro drug release profile in comparison to the marketed formulation (Larither®). Comparative in vivo anti-malarial performance of the developed SMEDDS was evaluated against the (Larither®) in Swiss male mice infected with lethal ANKA strain of Plasmodium berghei. The parameters studied were percent parasitemia, activity against time and animal survival period. Both the BAM-SMEDDS showed excellent self-microemulsification efficiency and released >98% of the drug in just 15 min whereas (Larither®) showed only 46% drug release at the end of 1 h. The mean globule size for optimized BAM-SMEDDS was <100 nm. The anti-malarial studies revealed that BAM-SMEDDS resulted in significant improvement in the anti-malarial activity (P < 0.05) as compared to that of (Larither®) and BAM solubilized in the oily phases and surfactant. The developed SMEDDS highlight safety for use and potential applications of indigenous natural lipophile in the development of novel colloidal drug carriers