Molecular Medicine

Poly (ADP-ribose) Polymerase (PARP) Inhibitor and its Role in DNA Repair and Treatment of Triple Negative Breast Cancer

by Stephanie Sandvick

This is my senior thesis review paper. My interest in PARP inhibitor was sparked by research I did at Stanford University.

Triple negative (TN) breast cancer accounts for 10% of all sporadic breast cancers and lack three specific cellular... more Triple negative (TN) breast cancer accounts for 10% of all sporadic breast cancers and lack three specific cellular receptors, estrogen receptor-α, progesterone receptor, and HER2 receptor/NEU oncogene (termed ER-, PR-, HER2). Interestingly, preclinical research has found that TN breast cancers are phenotypically similar to BRCA1 and BRCA2 mutated breast cancers. Research also suggests that the BRCA1 gene is involved in several types of DNA repair mechanisms including double-stranded break repair, nucleotide excision repair, and DNA cross-link repair and that mutations in BRCA1 result in defects in DNA repair. These mutations have been shown to be highly sensitive to chemotherapy drugs such as mitomycin C and cisplatin. Recently, it has been indicated that drugs called Poly (ADP-ribose) polymerase (PARP) inhibitors may have a primary role in treating BRCA mutated cancer. This review discusses the biological mechanisms of PARP and PARP inhibition as well as PARP inhibitor’s implications and limitations.

HLA CLASS II HAPLOTYPES ASSOCIATION IN RHEUMATOID ARTHRITIS, INSULIN-DEPENDENT DIABETES MELLITUS, MULTIPLE SCLEROSIS AND HASHIMOTO THYROIDITIS IN A GREEK POPULATION

by Andreas Anestis

Tissue Antigens, 2010 (Conference Abstract)

DRB1* subtypes associations in four autoimmune diseases in a Greek population

by Andreas Anestis

Tissue Antigens 2009 (Conference abstract)

Autoimmunity results in complex diseases by reason of many interacting
environmental and genetic components. In... more Autoimmunity results in complex diseases by reason of many interacting
environmental and genetic components. In addition, the complex nature of almost all
the class II associated autoimmune diseases includes polygenetic contributions from
HLA system.
We report the associations of HLA-DR4 subtypes among 4 autoimmune diseases:
rheumatoid arthritis (RA), insulin-dependent diabetes mellitus (IDDM), multiple
sclerosis (MS) and Hashimoto thyroiditis (HT). Distribution of HLA-DRB1*
genotypes, including DRB1*04 subtypes, was evaluated in four patient groups (RA,
HT, MS, IDDM) of 125 subjects each, and in 500 healthy control individuals by
PCRSSP.
Data were analyzed by chi-square test and Bonferroni method was used for statistical
correlations. Multivariate analyses (PCA, SCA) were performed for the identification
of grouping trends of genotypes. DRB1*04/03 frequency was higher than any other
genotype in all patient groups.
In each group concerning other DRB1* genotypes, DRB1*04 allele was combined -
HT: 11, 13, MS: 15, RA and IDDM: 04 homozygosity. In control group, compared to
each of the patient groups 11/07, 11/4 and 11/11 genotypes were found higher.
Concerning DRAˆ 1*04 subtyping, *0401, *0404 and *0405 subtypes were found in
HT,RAand IDDM, while inMS 0401 and 0403 were more frequent. In control group,
16/0403 and 11/0402 were significantly higher compared to all patient groups.
The multifactorial nature of autoimmune diseases makes the study of predisposing
factors problematic. The slight conformational differences in the binding sites of DR4
molecules could explain their differential participation in disease characteristics. The
identification of susceptibility genes may provide important markers for the early
identification of ‘‘high-risk’’ subjects.

GENETIC STUDIES OF DUCHENENE MUSCULAR DYSTROPHY AND PRIMARY MICROCEPHALY IN PAKISTANI POPULATION

by Saqib Mahmood

Intragenic Deletions In the Dystrophin Gene In 211 Pakistani Duchenne Muscular Dystrophy Patients

by Saqib Mahmood

A Recurrent Intragenic Deletion Mutation In DSG4 Gene In Three Pakistani Families With Autosomal Recessive Hypotrichosis

by Saqib Mahmood

Serum Melatonin Level and Oxidative Stress In Sickle Cell Anemia

by Edis Belini Junior

This study evaluated serum melatonin levels in patients with sickle cell anemia (SCA) and compared the results to... more This study evaluated serum melatonin levels in patients with sickle cell anemia (SCA) and compared the results to lipid peroxidation by determining thiobarbituric acid-reactive substances (TBARS) and Trolox equivalent antioxidant capacity (TEAC). The group studied was composed of 15 SCA patients and 24 subjects without hemoglobinopathies. The average melatonin level was significantly reduced in the SCA patients (p<0.001) when compared to the control group. The SCA patients showed significantly higher values for TBARS and TEAC when compared to values obtained for the control group (p<0.001 and p<0.01). Results from the correlation analysis in the SCA group were not statistically significant for any parameters except for TBARS and TEAC levels, which had a positive correlation (r=0.51; p=0.04), suggesting the participation of melatonin in antioxidant defense. The use of melatonin could be a possible therapeutic target for improving antioxidant defense and to reduce oxidative damage, alleviating symptoms associated with SCA

The Xmnl polymorphic site 5 ' to the gene G gamma in a Brazilian patient with sickle cell anaemia - fetal haemoglobin concentration, haematology and clinical features

by Edis Belini Junior

We report a 20-year-old female with sickle cell anaemia and with an HbF concentration of 15.8%. The patient was not... more We report a 20-year-old female with sickle cell anaemia and with an HbF concentration of 15.8%. The patient was not using hydroxyurea and was not receiving regular blood transfusions. The patient never had chronic manifestations of sickle cell anaemia, only pain crises of a mild intensity. After laboratory tests, we found that she was homozygous for HbS with the Bantu/atypical haplotype, and was heterozygous for the Xmnl site. The influence of the Xmnl site on the expression of HbF can explain the amelioration in clinical features in this haplotype association in a case of sickle cell anaemia

The profile of beta thalassemia obtained by data mining analysis in a database

by Edis Belini Junior

Thesis Abstract-Oxidative stress in sickle cell patients: influence of haplotypes and specific medication

by Edis Belini Junior

Einstellungen von Medizinstudentinnen und -studenten zu humangenetischer Forschung und genetischer Diagnostik

by Mike S. Schäfer

with  Weißflog, Gregor. 2005. German Medical Science. 22/2.

Metabolic markers of the head and neck cancers--clinical applications and the biochemical background

by Anna M. Czarnecka

The problem of diagnosis in the field of head and neck region is still valid. Specific diagnosis and precise... more The problem of diagnosis in the field of head and neck region is still valid. Specific diagnosis and precise estimation of the tumor's size with the use of CT and MRI imaging is generally unsatisfactory. The Positron Emission Tomography (PET) supports this process with additional information about the tumor's metabolism. Numerous publications show that PET-CT has a great influence on the evaluation of the size of the tumor, presence of lymph node metastases, choice of treatment and the prognosis of the recurrence. Cancer cells represent a specific metabolic state. These cells intake large quantities of glucose and utilize it in the process of glycolysis. The oxidative phosphorylation is not efficient in the transformed cells and defects in mitochondrial functions are at the heart of malignant cell transformation. Disruption of the oxidative phosphorylation chain has been described in the neoplasms. As a consequence, in cancer the glycolysis is active even in the normoxic environment. This metabolic shift in cell transformation has been described in early XX century and so called Warburg's hypothesis profoundly influenced the present perception of cancer metabolism, positioning what is termed aerobic glycolysis in the mainstream of clinical oncology. Today we know that neoplastic cells differ at the proteomic level. A subset of different proteins such as hexokinase II or HIF are upregulated. These abnormalities might be used as the neoplastic markers.

The impact of mtDNA mutations on proteins structure in selected types of cancer

by Anna M. Czarnecka

Recently published papers report a large number of mitochondrial DNA mutations in many different cancer types, but... more Recently published papers report a large number of mitochondrial DNA mutations in many different cancer types, but their significance for electron transport chain proteins remains unknown. This review covers structural mutations of mitochondrial genes, choosing prostate cancer, esophageal cancer and epithelioma as research models. As all mitochondrial genes encode subunits of the electron transport chain, the review focuses on the consequences of structural mutations on cell metabolism

Metabolic markers of the head and neck cancers--clinical applications and the biochemical background

by Anna M. Czarnecka

The problem of diagnosis in the field of head and neck region is still valid. Specific diagnosis and precise... more The problem of diagnosis in the field of head and neck region is still valid. Specific diagnosis and precise estimation of the tumor's size with the use of CT and MRI imaging is generally unsatisfactory. The Positron Emission Tomography (PET) supports this process with additional information about the tumor's metabolism. Numerous publications show that PET-CT has a great influence on the evaluation of the size of the tumor, presence of lymph node metastases, choice of treatment and the prognosis of the recurrence. Cancer cells represent a specific metabolic state. These cells intake large quantities of glucose and utilize it in the process of glycolysis. The oxidative phosphorylation is not efficient in the transformed cells and defects in mitochondrial functions are at the heart of malignant cell transformation. Disruption of the oxidative phosphorylation chain has been described in the neoplasms. As a consequence, in cancer the glycolysis is active even in the normoxic environment. This metabolic shift in cell transformation has been described in early XX century and so called Warburg's hypothesis profoundly influenced the present perception of cancer metabolism, positioning what is termed aerobic glycolysis in the mainstream of clinical oncology. Today we know that neoplastic cells differ at the proteomic level. A subset of different proteins such as hexokinase II or HIF are upregulated. These abnormalities might be used as the neoplastic markers.

Aggressive osteoblastoma of the sphenoid bone

by Anna M. Czarnecka

For osteoblastoma, with its predilection for the spinal column and appendicular skeleton, the skull is an unusual... more For osteoblastoma, with its predilection for the spinal column and appendicular skeleton, the skull is an unusual site, and paranasal sinus involvement is very rare. Herein, we report on a case in which the disease was located within the sphenoid bone. Tto the best of our knowledge, this is the 4th reported case of osteoblastoma with a sphenoid origin (1). We report an osteoblastoma of the sphenoid sinus in a 12-year-old girl who presented with exophthalmos. Ccomputed tomography (CTct) demonstrated an expansile lesion of the sphenoid which caused the orbital contents to be compressed and deviated to the right. In the magnetic resonance imaging scan, the lesion was found to invade the cranial base in the frontal and temporal region, approximating to the cavernous sinus and internal carotid artery on the right. Bilateral fronto-orbital craniotomy was performed. Histologically, the lesion was composed of proliferating osteoblasts along with vascular stroma. tumor was described as an aggressive osteoblastoma. In the follow-up CTct four months later, a pathological mass was observed in the area of the nasal septum, and a signal void was present on all sequences in the densely sclerotic areas. A second resection was performed. Tthe patient has been disease-free for 61 months. Herein, we present the diagnosis and management of this unusual lesion

Upon oxidative stress, the antiapoptotic Hsp60/procaspase-3 complex persists in mucoepidermoid carcinoma cells.

by Anna M. Czarnecka

Hsp60, a mitochondrial chaperonin highly conserved during evolution, has been found elevated in the cytosol of cancer... more Hsp60, a mitochondrial chaperonin highly conserved during evolution, has been found elevated in the cytosol of cancer cells, both in vivo and in vitro, but its role in determining apoptosis during oxidative stress (OS) has not yet been fully elucidated. The aim of the present work was to study the effects of OS on Hsp60 levels and its interactions with procaspase- 3 (p-C3) and p53 in tumor cells. NCI-H292 (mucoepidermoid carcinoma) cells were exposed to various concentrations of hydrogen peroxide (H2O2) for 24 hours. Cell viability was determined by Trypan blue and MTT assays. DNA damage was assessed by the Comet assay, and apoptosis was measured by the AnnexinV cytofluorimetric test. Exposure to increasing concentrations of H2O2 resulted in a reduction of cell viability, DNA damage, and early apoptotic phenomena. Hsp60, p-C3, p53, and p21 were assessed by Western blotting and immunocytochemistry before and after OS. Hsp60 and p-C3 were present before and after OS induction. Immunoprecipitation experiments showed an Hsp60/p-C3 complex before OS that persisted after it, while an Hsp60/p53 complex was not detected in either condition. The presence of wild type (wt) p53 was confirmed by RT-PCR, and p21 detection suggested p53 activation after OS. We postulate that, although OS may induce early apoptosis in NCI-H292 cells, Hsp60 exerts an anti-apoptotic effect in these cells and, by extension, it may do so in other cancer cells.

The value of immunohistochemical research on PCNA, p53 and heat shock proteins in prostate cancer management

by Anna M. Czarnecka

This review addresses the significance of the expression of proliferating cell nuclear antigen (PCNA), p53 and some... more This review addresses the significance of the expression of proliferating cell nuclear antigen (PCNA), p53 and some heat shock proteins (Hsps) in prostate carcinoma (PC). In fact, PCNA and p53 are two widely discussed tools in PC diagnosis, mainly because of the controversy regarding the significance of their expression during prostate cancer development and progression. At the same time, only few studies have shown the potential role of Hsps in carcinogenesis and their overexpression in pre-neoplastic and neoplastic lesions of the prostate. We briefly describe the physiological roles of Hsps in normal cells, and the significance of their immunohistochemical detection in PC as well as in pre-cancerous lesions of the prostate. We will also discuss the possible functional interactions of these molecules in both dysplastic and neoplastic cells.

Multi-Chaperones-Interactors Network in Mitochondria (MtCIN): Its Role in Carcinogenesis and Methodology of Analysis

by Anna M. Czarnecka

In human mitochondria, the characterization of multiple functional and structural interactions of mitochondrial... more In human mitochondria, the characterization of multiple functional and structural interactions of mitochondrial chaperones is needed for the understanding of the cellular protein biogenesis as well as of the process of carcinogenesis. In light of current
available data, we propose that mitochondrial Hsp70, Hsp60/10, DnaJ-like proteins, GrpE-like proteins, Clp/Hsp100, and Hsp90 families should be analyzed as a part of multi-chaperones-interactors network, and that single protein-protein interaction
analysis does not constitute sufficient explanatory framework of carcinogenic potential of mitochondrial chaperones. We also believe that new chaperones and cell-cycle regulatory interactors are to be defined in the near future, but many technical and
methodological efforts are needed before any generalization of the data will be possible and a reliable map of multi-chaperones-interactors network will be constructed.

Mitochondrial chaperones in cancer: from molecular biology to clinical diagnostics

by Anna M. Czarnecka

Mitochondria are cell organelles involved in processes of cell life and death, and therefore also in tumoral... more Mitochondria are cell organelles involved in processes of cell life and death, and therefore also in tumoral transformation. Indeed, mitochondria dysfunction is a prominent feature of cancer cells. Mitochondrial proteins and DNA have also been previously studied as markers of tumorigenesis. Heat shock proteins (HSPs) are ubiquitous evolutionary conserved proteins. HSPs enhance their expression in stressed cells and they are involved in gene expression regulation, DNA replication, signal transduction, differentiation, apoptosis, cellular senescence or immortalization. This review reflects recent views on the role of some mitochondrial molecular chaperones as prohibitin, mortalin and HSP60/HSP10 complex and their modifications leading to cell transformation and cancer development. These molecules could represent modern molecular biomarkers for oncological management.

CD1a down-regulation in primary invasive ductal breast carcinoma may predict regional lymph node invasion and patient outcome.

by Anna M. Czarnecka

AIMS: CD1a is a molecule belonging to the highly conserved group of CD1 proteins. Its expression in dendritic cells is... more AIMS: CD1a is a molecule belonging to the highly conserved group of CD1 proteins. Its expression in dendritic cells is related to the presentation of tumour-derived glycolipid antigens to T cells and, consequently, the development of a successful antitumour response. The aim was to investigate the presence of CD1a+ cells in both primary tumours and lymph nodes (LN) of a series of 35 invasive ductal carcinomas by both immunohistochemistry and reverse transcription-polymerase chain reaction. METHODS AND RESULTS: CD1a antigen was more expressed in N0 than N1 breast cancer (P < 0.0001) in both primary lesions and LN metastases and correlated positively and significantly with oestrogen (ER) (P = 0.0025) and progesterone (P = 0.0226) receptor (PR) status, as well as CD4+ and CD8+ T-lymphocyte infiltration. CONCLUSIONS: This is the first report to show a link between CD1a+ mononuclear cells in breast cancer and in paired LN metastases. The positive and significant correlations between the number of CD1a+ cells and positivity of the primary tumour for ER and PR suggest a possible role for CD1a as a prognostic marker for breast cancer, raising the possibility that hormone receptor-positive breast cancer patients may have a better prognosis in the presence of greater dendritic cell infiltration.