Prevalence of the MDR1 gene mutation in herding dog breeds and Thai Ridgebacks in Thailand

Background and Aim: A canine multi-drug resistance 1 (MDR1) nt230(del4) is a well-known inherited disorder that primarily affects collies and various herding breeds. The most recognized clinical implication for affected dogs is associated with an increased risk of multiple drug toxicity. To date, MDR1 gene mutations have been identified globally, especially in dogs from the USA and European countries. This study aimed to investigate the prevalence of MDR1 nt230(del4) in herding dog breeds and Thai Ridgebacks in Thailand. Materials and Methods: We clarified the prevalence of MDR1 nt230(del4) in 263 dogs of eight purebred dog breeds in Thailand using an allele-specific multiplex polymerase chain reaction method and direct DNA sequencing. Results: Rough Collies, Australian Shepherds, Shetland Sheepdogs, and Old English Sheepdogs were affected by the mutation with mutant allelic frequencies of 57.14%, 12.82%, 11.28%, and 8.33%, respectively. Among these populations, the prevalence of the MDR1 (+/–) genotype was 57.14% (12/21) for Rough Collies, 25.64% (10/39) for Australian Shepherds, 16.13% (15/93) for Shetland Sheepdogs, and 16.67% (2/12) for Old English Sheepdogs, whereas the MDR1 (–/–) mutation was only identified in Rough Collies and Shetland Sheepdogs, with prevalences of 28.57% (6/21) and 3.22% (3/93), respectively. However, the MDR1 nt230(del4) was not identified in Border Collies, German Shepherds, White Swiss Shepherds, or Thai Ridgebacks. Conclusion: This study provides the current situation regarding MDR1 nt230(del4) in herding dog breeds in Thailand. In this survey, we investigated for the first time the status of MDR1 genotype in Thai Ridgebacks. These results are helpful for veterinarians managing effective therapeutic plans for commonly affected dog breeds, and these results will encourage all breeders to improve their selective breeding programs based on the MDR1 nt230(del4) status.

High resolution melting analysis and detection of Leishmania resistance: the role of multi drug resistance 1 gene

Background Pentavalent antimonial compounds are currently used to treat leishmaniasis and resistance to these drugs is a serious problem. Multidrug resistance protein is an efflux pump of the cell membrane that expels foreign compounds. This study designed to evaluate the mutations in the multi-drug resistance 1 (MDR1) gene, in biopsy specimens of Leishmania tropica, with high resolution melting (HRM) method. In this experimental study, genomic DNA was extracted from 130 patients with skin leishmaniasis. Then, nucleotide changes were investigated throughout the gene using HRM and sequencing methods. The samples categorized in 5 groups by differences in the melting temperature (Tm). Result The nucleotide changes analysis showed that 61% of the samples of different groups that were unresponsive to drug had mutations in the MDR1 gene, which were also confirmed by the sequencing method. These mutations can be one of the factors responsible for non-responsiveness to the treatment. Conclusion According to the findings, it seems that mutation in MDR1 gene could be responsible for drug resistance to pentavalent antimonial compounds. Furthermore, HRM method can be used to diagnose drug resistance in leishmaniasis. It is also recommended that further studies be done regarding the importance of drug resistance in the leishmania affected patients.

Nucleolin Promotes Cisplatin Resistance in Cervical Cancer by the YB1-MDR1 Pathway

Purpose. Cervical cancer is the fourth most common cancer in women worldwide and is the main cause of cancer-related deaths in women. Cisplatin (DDP) is one of the major chemotherapeutic drugs for cervical cancer patients. But, drug resistance limits the effectiveness of cancer therapy. Nucleolin (NCL) is a nucleocytoplasmic multifunctional protein involved in the development of cancer. It has been reported that NCL may be a potential target for modulation of drug resistance. However, the precise molecular mechanisms are poorly understood. Materials and Methods. Human cervical cancer Hela cells and their cisplatin-resistant cell line Hela/DDP were used in this study. The protein level of NCL in cervical cancer cells was measured by western blot analysis. Hela cells and Hela/DDP cells were transfected with NCL overexpression plasmid or NCL siRNA separately. MTT and EdU assay were performed to evaluate the cell viability and sensitivity to cisplatin. The drug efflux function of MDR1 protein was assessed by intracellular rhodamine-123 accumulation assay.The promoter activity of MDR1 was assessed by using a dual-luciferase reporter assay. Results. We found that the protein level of NCL was elevated in Hela/DDP cells. Overexpression of NCL increased cervical cancer cell proliferation and attenuated the sensitivity to cisplatin. Overexpression of NCL increased Multidrug resistance (MDR1) gene expression and drug efflux. Our results demonstrated that NCL was highly related with cisplatin resistance in cervical cancer. NCL played an important role in MDR1 gene transcription through regulation of the transcription factor YB1. Conclusion. Our findings revealed the novel role of NCL in cisplatin-resistant cervical cancer and NCL may be a potential therapeutic target for chemoresistance.

First report of nt230(del4) mutation in the MDR1 gene in German Shepherds in Southern Brazil

ABSTRACT: The P-glycoprotein (P-gp) is a transmembrane protein encoded by the MDR1 gene that functions as a biological barrier by extruding toxins and xenobiotics out of cells. The MDR1 gene can carry a mutation called nt230(del4), which is a deletion of four base pairs resulting in the formation of a non-functional protein that may predispose to severe toxicosis, as observed in dogs with sensitivity to ivermectin. Several breeds have been described as carriers of the mutation, including German Shepherds (GS). However, the presence of the mutant allele in this breed has not been described in Brazil. This study aimed to determine the genotypic and allelic frequency of the nt230(del4) mutation in the MDR1 gene in GS from Southern Brazil. Blood samples were collected from 79 GS in the state of Rio Grande do Sul and genotype for the MDR1 gene was performed. Seventy-eight (98.7%) dogs were dominant homozygous genotype (wild) and one (1.3%) was heterozygous. This study showed that there is a low frequency (0.6%) of the mutant allele while the frequency of the wild allele is high (99.4%) in this specific population. This is the first report of the presence of the nt230(del4) mutation in the MDR1 gene in GS in Brazil. This information is important for breeders to prevent dissemination of the mutant allele in the national breeding population and international exchange of animals for breeding; for owners and veterinarians to be aware when dispensing and administering medications for GS dogs in Brazil.

High Resolution Melting Analysis in Leishmania Resistance: The Role of Multi Drug Resistance 1 Gene

Objective Pentavalent antimonial compounds are currently used to treat Leishmaniasis. Resistance to these drugs is a serious problem. The purpose of this study was to evaluate the mutations in the multi-drug resistance 1 (MDR1) gene in biopsy specimens of Leishmania Tropica with high resolution melting (HRM) method.Results In this experimental study, genomic DNA was extracted from 130 patients with skin Leishmaniasis. Then, nucleotide changes were investigated throughout the gene length using HRM and sequencing methods. The results of the nucleotide changes showed that 61% of the samples that were unresponsive to drug had mutations in the MDR1 gene in different groups, which were confirmed by the sequencing method. These mutations can be one of the factors responsible for non-response to the treatment of the disease. HRM method can be used to diagnose drug resistance in Leishmaniasis. It is also recommended that further studies be done regarding the importance of drug resistance in the affected patients.

Genetic evidences for the core biosynthesis pathway, regulation, transport and secretion of liamocins in yeast-like fungal cells

So far, it has been still unknown how liamocins are biosynthesized, regulated, transported and secreted. In this study, a highly reducing polyketide synthase (HR-PKS), a mannitol- 1-phosphate dehydrogenase (MPDH), a mannitol dehydrogenase (MtDH), an arabitol dehydrogenase (ArDH) and an esterase (Est1) were found to be closely related to core biosynthesis of extracellular liamocins in Aureobasidium melanogenum 6-1-2. The HR-PKS was responsible for biosynthesis of 3,5-dihydroxydecanoic acid. The MPDH and MtDH were implicated in mannitol biosynthesis and the ArDH was involved in arabitol biosynthesis. The Est1 catalyzed ester bond formation of them. A phosphopantetheine transferase (PPTase) activated the HR-PKS and a transcriptional activator Ga11 activated expression of the PKS1 gene. Therefore, deletion of the PKS1 gene, all the three genes encoding MPDH, MtDH and ArDH, the EST1, the gene responsible for PPTase and the gene for Ga11 made all the disruptants (Dpks13, Dpta13, Dest1, Dp12 and Dg11) totally lose the ability to produce any liamocins. A GLTP gene encoding a glycolipid transporter and a MDR1 gene encoding an ABC transporter took part in transport and secretion of the produced liamocins into medium. Removal of the GLTP gene and the MDR1 gene resulted in a Dgltp1 mutant and a Dmdr16 mutant, respectively, that lost the partial ability to secrete liamocins, but which cells were swollen and intracellular lipid accumulation was greatly enhanced. Hydrolysis of liamocins released 3,5-dihydroxydecanoic acid, mannitol, arabitol and acetic acid. We proposed a core biosynthesis pathway, regulation, transport and secretion of liamocins in A. melanogenum.