Title: Molecular Cloning of TUL4 gene of Francisella tularensis

Abstract:

Francisella tularensis is a pathogenic gram-negative bacterium and causes a pneumonic type of often fatal disease called Tularaemia. F. tularensis can infect a large number of mammalian species including humans. It is a fastidious, facultative intracellular bacterium, which requires cysteine for growth. Tularemia is treated with anti-microbials, for example, aminoglycosides, antibiotic  medications, or fluoroquinolones. The objective of the assignment is to clone a PCR target gene (TUL4) of F.tularensis in E.coli. PCR was performed using genomic DNA of F. tularensis as a template. After PCR, the amplicon was purified and cloned into a cloning vector. Positive clones were selected based on colony PCR screening. Recombinant plasmids from the positive E. coli clones were purified and kept for subsequent downstream applications.

Title:

Abstract:

Gene cloning or molecular cloning is an important step in recombinant DNA technology wherein a sequence of one or more DNA fragment is introduced into another DNA sequenced for the purpose of amplification or functional studies. It is also used for downstream applications such as sequencing, mutagenesis, genotyping or heterologous expression of a protein. The traditional techniques of gene cloning involve the transfer of DNA fragment of interest from one organism to a self-replicating genetic element such as a bacterial plasmid. The process involves amplification of the desired gene fragment, splicing of the gene with restriction enzyme, if required and ligating into a plasmid. The final assembly includes promoter element fused to a reporter gene under the control of a ubiquitous promote.  The assembly is then  inserted  into  a host,  which  may  be  an eukaryotic or a prokaryotic cell. Gene cloning is important to many areas of modern biology and medicine.

Borrelia is a spirochete bacterium which cause Lyme Disease in humans. Transmission to human is caused by ticks of Ixodes spp. Several species of Borrelia cause Lyme disease some of which are B. burgdorferi, B. afzelii and B. garinii. Of these B. burdorferi is the most important and the most studied cause of Lyme Disease. The disease is worldwide in distribution including India (Vasudevan and Chatterjee, 2013; Praharaj et al., 2008). B. burgdorferi enters the skin at the site of the tick bite, the organism then migrate locally in the skin around the bite, spread via the lymphatics to cause regional adenopathy or disseminate in blood to organs or other skin sites. Clinical symptom may include fever, chills, headache, fatigue, muscle, joint aches and erythema migrans rash.  For effective treatment, early diagnosis is of paramount importance. Among the many techniques employed for diagnosis of Lyme Disease in the past, molecular detection using PCR remains the most rapid and sensitive test (Marconi and Garon, 1992)

The present work involved cloning of a segment of a 16S rRNA gene of Borrelia burgdorferri for further characterization and downstream applications.  The process involves PCR amplification of a fragment of the 16S rRNA gene of Borrelia burgdorferi, ligating into a plasmid vector and transformation into an E. coli host.

Title: Energy Alterations In Patient with Fragile X Syndrome

Abstract:

Fragile X syndrome is a gentic condition caused by less FMR1 gene (which is on chromossome X), that is a protein that helps nerve cells “ talk” to one another. Thi slack of FMR1 gene can affect child’s learning, health, appearance and behaviour. The purpose of this study is to demonstrate that children with Fragile X Syndrome have energy deficinecy in the five internal massive organs ( LIver, Heart, Spleen, Lung and Kidney) , according to traditional Chinese medicine’s theories, and the treatment of this condition can improve the quallity of life of these patients and improve their symptoms. Methods: through one case report of a 14 year-old male patient that went to my clinic due to fear, diffucult in speech, problems in the school due to difficult in learning, anxiety. In his physical examination, there had a big ears, a long thin face and pectus excavatum. He had a diagnosis of Fragile X syndrome after investigation on the São Paulo University when he was yourger. I did the radiesthesia procedure to evaluate his energy situation in the internal five massive organs. Results: what I found was that all his internal organs were in the lowest level of energy, rated one out eight, with exception of the seventh chakra, that was normal, rated in eight. The treatment consisted in Chinese dietary counseling, auricular acupuncture with apex ear bloodleting and it was used homeopathy medications according to the theory of the Constitutional Homeopathy Five Elements Based on Traditional Chinese Medicine and crystal-based medications ( to replenish the energy of the five internal massive organs). After the first session of treatment, the patient started to fell much better, less anxiety and fear, more happy and smiling every time he saw me, and he also started to talk more and comunicate better with family members and in the school. Conclusion: the conclusion of this study is that patient with Fragile X syndrome have energy alterations in the five internal massive organs and the treatment of this condition, replenishing these energies using highly dilluted medications ,together with tools used to balance the state of Yin, Yang, Qi and Blood using Chinese dietary counseling , auricular acupuncture with apex ear bloodleting is of major importance to improve the quality of life of these patients. Studies using more patient with this same problem will be important to prove this findings.

Biography:

Huang Wei Ling, born in Taiwan, raised and graduated in medicine in Brazil, specialist in infectious and parasitic diseases, a General Practitioner and Parenteral and Enteral Medical Nutrition Therapist. Once in charge of the Hospital Infection Control Service of the City of Franca’s General Hospital, she was responsible for the control of all prescribed antimicrobial medication and received an award for the best paper presented at the Brazilian Hospital Infection Control Congress in 1998. Since 1997, she works with the approach and treatment of all chronic diseases in a holistic way, with treatment guided through the teachings of Traditional Chinese Medicine and Hippocrates. Researcher in the University of São Paulo, in the Ophthalmology department from 2012 to 2013. Author of the theory Constitutional Homeopathy of the Five Elements Based on Traditional Chinese Medicine. Author of more than 100 publications about treatment of variety of diseases rebalancing the internal energy using Hippocrates thoughts.