Saturday, January 25, 2020

ERG11 in Drug-resistant of C. Krusei

ERG11 in Drug-resistant of C. Krusei Erg11 mutations and up-regulation in clinical itraconazoleresistant isolates of Candida krusei Some C. krusei isolates were resistant the antifungal drugs. Mutations including T939C, T642C and A756T were found in ERG11 of C. krusei. ERG11 was highly expressed in resistant C. krusei strains.. Abstract Objective We aimed to provide with light for better understanding of ERG11 gene in drug-resistance mechanisms in Candida krusei (C. krusei). Methods C. krusei strains were isolated from patients in the Dermatology from Jan 2010 to May 2013. Susceptibility assays, including 5-fluorocytosine (5-FC), amphotericin B (AMB), voriconazole (VCR), fluconazole (FLC) and itraconazole (ITR), was performed by broth microdilution method according to the National Committee for Clinical Laboratory Standards M27-A2. Isolates were divided into susceptible strains and resistant strains based on their susceptibility to ITR. Mutations in the ERG11 gene sequence were detected using PCR amplification and gene sequencing. Expression levels of ERG11 were measured by real-time PCR. Differences of ERG11 expression levels between susceptible strains and resistant strains were compared by two-tailed Student t test. Results A total of 15 C. krusei strains were obtained, among which 8.0, 6.0 and 3.0% were resistant to FCA, ITR and 5-FC, respectively, whereas all isolates were found to be susceptible to AMB and VRC. Three synonymous codon substitutions were found in ERG11among all the C. krusei strains, including T939C, T642C and A756T. Expression level of ERG11 was significantly higher in resistant C. krusei strains (1.34  ± 0.08) than that in susceptible C. krusei strains (0.94  ± 0.14) (t = 3.74, P Conclusions Our study demonstrates that point mutations accompanied with the overexpression of ERG11 might be involved in the molecular mechanisms of drug resistance in C. krusei. Keywords: ERG11; drug resistance; Candida krusei; mutation; overexpression. Introduction Candida species are pathogenic microorganisms for systemic and local opportunistic infections and the fourth leading cause of nosocomial bloodstream infections worldwide [1]. In recent years, an increasing number of infections due to Candida krusei has been witnessed [2]. C. krusei generally causes infections among immunocompromised patients, especially those suffering from Human immunodeficiency virus (HIV)-acquired immune deficiency syndrome and hematological malignancies [3]. Mortality rates among such patients with C. krusei fungemia are reported to be as high as 60-80% [4]. Moreover, the increasing use of immunosuppressive drugs has inevitably increased the risk of C. krusei infection, and C. krusei has ranked in fifth place among all the species of Candida [5, 6]. C. krusei has been regarded as a multidrug-resistant (MDR) fungal pathogen by the reason of its intrinsic resistance to fluconazole (FLC) as well as its considerable reduction in susceptibility to flucytosine and amphotericin B (AMB) [2]. Resistance to these antifungal drugs is a major problem among patients with acute myelogenous leukemia, neutropenia and/or critically ill, since these drugs are frequently used for prophylaxis of C. krusei infection [7]. Therefore, a better understanding of resistance mechanisms in C. krusei was urgently needed to effectively prevent and control infections caused by C. krusei. It has been reported that multiple mechanisms are involved in drug resistance in C. krusei , including overexpression of several genes such as multidrug transporters (encoded by CDR1, CDR2, and MDR1), which lead to decreased intracellular accumulation of FLC. Nowadays, an increasing evidence proposed that alterations and overexpression of the ERG11 gene, which codes the target enzyme cytochrome P450 lanosterol 14ÃŽ ±-demethylase, are major resistance mechanism in C. krusei [8, 9]. However, the scarce information available regarding ERG11 gene mutation and gene expression in the drug-resistant strains makes C. krusei resistance to azoles poorly understood [10]. In the present study, we evaluated the susceptibility profiles of 15 C. krusei isolates, investigated the potential alterations in the ERG11 gene sequence of C. krusei strains, and further detected the differentially expressed levels of ERG11 between susceptible and resistant isolates of C. krusei. We aimed to provide with light for better understanding of molecular mechanisms in drug resistance of C. krusei. Methods 2.1 Fungal strains and media The strains used in this study were isolated from patients in the Dermatology of the Second Hospital of Shanxi Medical University from Jan 2010 to May 2013. The standard strain, Candida krusei ATCC 6258, was purchased from fungi and fungal disease research center of Peking University and included in each test run for quality control. C. Krusei were maintained on agar YPD medium (2% peptone, 1% yeast extract, 2% dextrose) and stored in our Fungi Laboratory. RPMI 1640 medium with L-glutamate (Sigma, St. Louis, Mo.) was used as recommended for susceptibility assays and buffered to pH 7.0 with 0.165 M morpholinepropanesulfonic acid (MOPS). 2.2 Susceptibility assays The susceptibility assays of the isolates was performed in 96-well polystyrene microtiter plates by broth microdilution method described in the National Committee for Clinical Laboratory Standards M27-A2 (NCCLS) [11]. The antifungal drugs including 5-fluorocytosine (5-FC), amphotericin B (AMB), voriconazole (VCR), fluconazole (FLC) and itraconazole (ITR), were obtained from their respective manufacturers and used for susceptibility assays. MIC was defined as the concentration of the drug that reduced the fungus growth by 80% compared to that grown in the absence of the drug. The interpretive criteria for susceptibility to the above drugs were published by the NCCLS and listed in Table 1. Samples were divided into susceptible strains and resistant strains based on their susceptibility to ITR. 2.3 PCR amplification and sequence alignment of ERG11 gene To amplify ERG11 gene, genomic DNA was firstly isolated from C. krusei cells using UNIQ-10 Column Genomic DNA Isolation Kit (Sangon Biotech, Shanghai, China) according to the manufacturer’s instruction and used as a template for amplification of the ERG11 genes. Specific primers of ERG11 (Table 2) were designed by Primer 3 [12], based on the available sequence information of C. Krusei ERG11 gene (GI:163311561) at the National Center for Biotechnology Information (NCBI). The PCR amplification of ERG11 gene was conducted using 2  µl of genomic DNA, 2  µl specific forward and reverse primers (50  µmol/L) and Taq PCR Master Mix (TIANGEN, Beijing, China). The PCR condition was set as denaturation for 5 min at 94  °C, followed by 35 cycles: 94  °C for 30 s, 55  °C for 30 s and 72  °C for 30 s, and a final step of elongation (72  °C for 8 min). PCR products were then separated and sized on a 1.5% agarose gel by electrophoresis, and visualized under UV light after stai ning with ethidium bromide. Successfully amplified PCR products were sent for sequencing (Invitrogen, Shanghai, China). To verify the point mutations, sequencing results were aligned with the reference sequence of C. Krusei ERG11 gene (GI:163311561) using BLAST (Basic Local Alignment Search Tool) program in NCBI. 2.4 Real-time PCR analysis For quantitative real-time PCR analysis, total RNA was extracted from C. krusei cultures with an Yeast RNAiso Kit (TaKaRa, Dalian, China) and reversely transcribed to cDNA with PrimeScript RT Master Mix (TaKaRa, Dalian, China) according to the instructions of the manufacturer. For the ERG11 target genes and GAPDH reference gene, a primer pair were designed with Primer 5.0 program (Table 2). Real-time PCR was processed with a 25- µl volume containing the following reagents: 12.5  µl of SYBR ® Premix Ex TaqTM II (TaKaRa, Dalian, China), 2  µl of total RNA sample, 1 µl of each primer pair at a concentration of 10  µM and distilled water up to the final volume. Samples were subjected to an initial step at 95  °C for 5 min, followed by 40 cycles each of which consisted of 10 s at 95  °C and 30 s at 60  °C. Melting curves were recorded every 5 s during the 65-95  °C by PCR amplifier. Fluorescence data (Ct) in each reaction were collected and were analyzed with the Rotor -Gene Q Series Software 2.0.2 software. A 2à ¢- ³Ãƒ ¢- ³Ct algorithm was applied to analyze relative expression levels of ERG11 at susceptible strains and resistant strains. 2.5 Statistical analysis Statistical analysis was performed using SPSS 17.0 software (SPSS incà ¯Ã‚ ¼Ã…’Chicagoà ¯Ã‚ ¼Ã…’US). All data were presented as mean  ± standard deviation (SD). The two-tailed Student t test was conducted to compare the differences of ERG11 expression levels between susceptible strains and resistant strains. A p-value Results Antifungal susceptibilities of C. krusei isolates A total of 15 C. krusei strains were isolated from clinical samples during Jan 2010 to May 2013 in our laboratory, among which 14 isolates were from urine and 1 from hydrothorax (Table 3). Table 4 shows the rates of azole resistance for these C. krusei isolates. Among 15 C. krusei isolates, 8.0, 6.0 and 3.0% were resistant to FCA, ITR and 5-FC, respectively, whereas all isolates were found to be susceptible to AMB and VRC. Ultimately, by using an MIC ≠¥ 1  µg/ml to define resistance to the investigational ITR, the 15 isolates included 6 that were resistant and 9 that were susceptible. Mutational analysis in ERG11of C. krusei isolates DNA fragment with consistent length was successfully amplified from C. krusei isolates (Figure 1). Sequence alignment showed three synonymous codon substitutions in ERG11among all the C. krusei strains, including T939C, T642C and A756T (Figure 2). Among the three synonymous mutations, T642C and A756T only presented in ITR-resistant strains, while T939C in susceptible as well as resistant C. krusei strains. Nevertheless, no point mutation was observed in the standard strain. Different expression levels of ERG11gene in susceptible and resistant C. krusei strains Real-time PCR was performed to verify the varied expression levels of ERG11 in ITR-susceptible and ITR-resistant C. krusei strains. Result showed that mRNA transcription level of ERG11 was significantly higher in resistant C. krusei strains (1.34  ± 0.08) than that in susceptible C. krusei strains (0.94  ± 0.14) (t = 3.74, P Discussion With the drug-resistance character, C. krusei has emerged as one of the leading agents causing candidemia, especially in immunocompromised patients [13]. Previous studies have shown the crucial role of ERG11 gene in FLC-resistant clinical isolates of Candida species, while the molecular mechanism specially in C. krusei remains unclear. Herein, we evaluated the susceptibility profiles of 15 C. krusei isolates, subsequently searched for mutations in the ERG11 gene sequence of C. krusei using PCR amplification and gene sequencing, and further detected the differentially expressed levels of ERG11 between susceptible and resistant isolates by real-time PCR. We found three synonymous codon substitutions in ERG11 of C. krusei which have not yet been described previously. Among the three point mutations, T642C and A756T only presented in ITR-resistant strains, while T939C were also presented in ITR-susceptible strains. Moreover, mRNA transcription level of ERG11 was significantly higher in I TR-resistant C. krusei strains than that in ITR-susceptible C. krusei strains. It is reported that point mutations in the ERG11 gene can lead to conformational changes which may affect the affinity of the drug with the target, but not influence the enzyme function in ergosterol biosynthesis [14]. Many studies have been focused on the ERG11 alterationz in Candida species . Ricardo et al. [9] found two different types of mutations by sequencing the C. krusei ERG11 gene, including a heterozygous alteration at 1,389 bp (T→C) presented in all of the susceptible and resistant C. krusei strains in their study, and a missense mutation in two strains at position 418 bp (T→C) which yields a Tyr→His amino acid change. Tavakoli et al. [15] displayed a heterozygous polymorphism at position 939 (T→C) in ERG11 coding region, and speculated that this polymorphism might play a key role in the transcriptional regulation of genes and be involved in the processes of ergosterol biosynthesis. Sionov et al. [16] have indentified a single missense mutation at amino acid 145 in the ERG11 of C. neoformans strain isolated from an FLC-treated patient, and verified that this mutation was sufficient to lead high FLC resistance. In the present study, we discovered three novel synonymous codon substitutions in ERG11 of C. krusei, among which T642C and A756T only presented in ITR-resistant strains. These alteration in ERG11might be involved in the resistance mechanism of C. krusei. Many studies have also been designed to explore the exact molecular mechanism behind the ERG11 up-regulation in response to azoles and other antifungal drugs [14, 17, 18]. A well-characterized matched pair of FLC-susceptible and FLC-resistant C. albicans isolates was analyzed and the resistant strains were found mainly associated with up-regulation of ERG11 gene [14]. Henry et al. [17] demonstrated that treating C. krusei with the triazole FLC at a concentration of 2 to 9 mg/ml could resulted in four- to five-fold increase in ERG11 RNA levels. Accompanied with previous reports, they hypothesized that the the upregulation of ERG11 gene contributed at least partly to the ability of C. krusei to tolerate azole [19-21]. The overexpression of ERG11was also directly shown to confer FLC resistance in S. cerevisiae [18]. In accordance with previous studies, our results reflected that mRNA transcription level of ERG11 was found to be significantly higher in ITR-resistant C. krusei strains com pared with that in ITR-susceptible C. krusei strains. The increased production of 14a-demethylase was reported to exceed the inhibitory capacity of the antifungal drugs. Therefore, we speculated that ERG11 gene overexpression might be relevant in the drug resistance in C. krusei. In conclusion, there synonymous codon substitutions were observed in ERG11 of C. krusei. These point mutations accompanied with the overexpression of ERG11 might be involved in the molecular mechanisms of drug resistance in C. krusei.

Friday, January 17, 2020

Lateralization of Functions: Left and Right Hemispheres

Brain is the most complex structure ever evolved in Universe such that it contains billions and billions of neurons that act and react in interconnected ways leading to the emergence of thought, consciousness, feelings, emotions and creative thinking, all subsumed under the enigmatic term mind. The brain is a multilayered structure that is folded into many folds, the surface of which is known as the gray matter and the inner areas deep within the cortex is known as the white matter (the color is because of the fatty substance).The cerebral cortex is the area of associative functions of the brain that is divided into many fissures such as lateral fissure and central fissure. The four lobes formed based on these fissures are the frontal lobes, parietal lobes, temporal lobes and occipital lobes. The cerebral hemisphere is divided into two halves: the left hemisphere and the right hemispheres with asymmetric functions.Corpous callousum is the structure that joints the two hemispheres so that the brain functions according to the Lashley principle of mass action, that is, it is the integrative functions of the two hemispheres that result in the coordinated activates of individuals, that is behavior. Lateralization of Functions: Left and Right Hemispheres Lateralization of brain function means that each hemisphere, that is the right or the left, specializes in certain functions so that the inactivity of a hemisphere result in the damage to that function or that function totally disappears from the behavior repertoire.Left hemisphere or right hemisphere dominances are responsible for the controlling of those respective functions. Lateraisation of functions may be studied using different methods like the ancient method of inserting an electrode in the brain and the modern methods of imaging, scanning and intravenous injections. The Wada Test introduces an anesthetic to one hemisphere of the brain, that is intravenous injection, following which, that is after anesthetize d, neuropsychological tests are conducted to determine the effect of hemispheric paralysis or inactivity.Less invasive techniques such as Magnetic Resonance Imaging, Computerized Axial Tomography, SQUID and Poisson Emission Tomography are used to study dominance. Brain function lateralization is also determined in the phenomena of right or left handedness and of right or left ear preference (Lateralization of brain functions, 2009). However it is to be stated that preferences are not always a clear indication of hemispheric specialization. Ninety five percent of the right handed people have left hemisphere dominance for language and only eighteen point eight per cent of left handed people have right hemisphere dominance for language functions.Sodium amytal tests indicate that 95 percent of right handed people are specialized in left hemisphere for language functions (Milner, 1974). Left hemisphere is specialized for language functions (Corina, et al, 1992). Left hemisphere strokes t hat leads to right sided paralysis (the left side of the brain controls the right side of the body and right side of the hemisphere controls the left side of the body known as ipsilateral control) result in serious language problems .The left hemisphere temporal lobe region known as the Wernicke’s area is involved in the understanding of spoken and written language. The language region in the lower frontal lobe is known as Broca’s area, the area involved in speech production The damage to the Broca’s area leads to the slow and infrequent speech thereby the fluency is impaired. The left brain does all the functions related to analysis, logical, mathematical thinking, cause-effect relations and scientific thinking. In figurative language, the left brain is said to be, the Western brain whereas the right brain is the Eastern brain.The sequential nature of processing of information carried out by the left hemisphere is contrasted with the global processing of right hemisphere (Springer and Deutsch, 1989). Right hemisphere specializes in â€Å"soft† functions characterized as appreciation of music, art, production of relaxation, quietness, peace, spatial relations, recognition and memory patterns of stimulation (Morgan, etal. 1993) and the patterns are visual, tactile or spatial. Damage to these areas prevents these functions so that the person may fail to recognize the face of other people, fail to appreciate music and fail to relax.People practicing mediation is said to show changes in the electrical activity of the brain which means that electrical waves called alpha dominate in their lives. Significant changes in the electrical activity of people practicing mediation and not practicing mediation are located in studies. Emotional expression is another function that dominates the right hemisphere (Zaidel, 1994). In short, identification of faces, facial expressions of emotions, line slopes or dot locations occur more quickly when these are ‘shown’ to the right hemisphere. The distinct functions are explained in Table 1.Split Brain Studies Different functional specialization of the hemispheres are observed in the studies of split brain patients, who are characterized by the severing of corpus callousum so that there is no anatomical connection between the two halves and the two hemispheres act independently. Roger S. Sperry has conducted many studies with split brain patients whose brain functioned independently without transferring message between the two halves. With split brain patients visual input can be restricted to the left or right hemisphere because of the anatomical connections between the eyes and the brain.From the right visual field, stimuli go to the left hemisphere and the input from the left visual field goes to the right hemisphere. In this way the visual image of the word ‘ring’ reaches the left hemisphere, while the word ‘key’ reaches the right hemisphere in a form of experiment. It is found that split brain patients report the word ‘ring’ and they do not report the word ‘key’ sent to the right hemisphere, implying that there is no connection between the two halves of the brain. Moreover the left hemisphere specializes in the language function. And it is also found that the patients can recognize the word â€Å"key† by non-verbal language.

Thursday, January 9, 2020

The Cultural Practices Of Buddhism - 1460 Words

In a general ranking of all the major religions in the world, Buddhism is a top contender. Practiced mainly in the eastern hemisphere and originating from ancient India, it has a following of hundreds of millions of people. Founded by a young man by the name of Siddhartha Gotama from Lumbini, now called Nepal, it characterises itself as a religion of truth and morality. Siddhartha Gotama, now referred to as Buddha, taught many things; however, the main principles of Buddhism can be summarized into the Law of Karma, the Four Noble Truths and the Noble Eightfold Path. With major strides in globalization in the last few decades, the teachings and principles of Buddhism have become more familiar to western cultures. The increase in the exposure to Buddhism has also been accompanied by a greater effort to be culturally aware and ultimately respectful to practicing Buddhists. In healthcare, where there is a wide cross section of individuals, the need for cultural awareness exists even more . This paper will discuss some of the cultural practices of Buddhists, as it relates to their beliefs, and the impact they have on the wellness and illness or Buddhists. It will also show how the ignorance of cultural differences in Buddhism can be remedied by cultural competency which would be expressed in the form of encourage therapeutic care. In Buddhism, the law of Karma is a law of cause and effect. According to Buddhism, everything that exists must have a cause (Rodgers Yen, 2002).Show MoreRelatedChina And Jap Spirituality, Belief And Faith Are Personal1271 Words   |  6 PagesStudy of Religion in China and Japan Spirituality, belief and faith are personal. This statement means that an individual takes his own path on religion. 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According to (https://www.google.com/#q=buddhismBuddhism) is a non-theistic religion or philosophy that encompasses a variety of traditions, beliefs and spiritual practices largely based on teachings attributed to the Buddha. Buddhism inhabits several traits that consider it a religion and a philosophy. There is not a clear distinction that defines buddhism as one or the other. Some practices of Buddhism display

Wednesday, January 1, 2020

The Argument On The Existence Of God - 1629 Words

The idea of God has been a part of man’s history for centuries. Since time began there has been various combination of believers, and non-believers. Individuals who believes in God, belong to many different religion. Whereas, skeptics find the existence of God somewhat baffling, and have continually sought answers to His existence through scientific methods. As the world progresses in scientific, and technological advancement, the human race still faces the question of God’s existence. Many philosophers, and scholars, such as Aquinas, Anselm, and Rene Descartes has applied their theory, in an effort to answer the question. Word.50. In this paper I will argue in favor of Descartes’ argument on the existence of God from his writings, Meditations of First Philosophy. I find that the following claim of Descartes’ ontological argument existence of God (clear and distinct idea of a supremely perfect being) provides convincing argument. The validity of the argument support his claim of the existence of God, for the following reasons. The idea of God is consistent with Scripture. Colossians 1:16 â€Å"For by Him all things were created; things in heaven and on earth, visible and invisible, whether thrones or dominion, principalities or powers, all things were created by Him and for him.† Our belief in the existence of God has enormous implications on our views of life, morality, humanity, and destiny. In the misinterpretation of Descartes’ argument, of necessary existence, some haveShow MoreRelatedThe Arguments For The Existence Of God1056 Words   |  5 Pages16 November 2015 Rough Draft for The arguments for the Existence of God. The question Does God Exist? is a well-known asked question in the world. Most people believe they know the answer to it. The religious people would say, well of course he does, while the non-religious people or atheist would say no He does not exist. Because evil exist and chaos exists, God cannot be all-powerful. In the modern world, there are many different opinions as to whether a God exists or not. This has been an issueRead MoreThe Argument For The Existence Of God1674 Words   |  7 Pagesfind the three arguments I analyzed satisfactory for the existence of God. The existence of God simply cannot be proven. Regardless of how strong a person’s faith is, or how many miracles they claim to have witnessed, God can only ever be a possibility. First, I will discuss why Pascal’s wager is not a satisfying argument for the existence of God. I will then examine C.D. Broad’s â€Å"Argument for the Existence of God†, and why it is also not a satisfying argument for the existence of God. Finally, IRead MoreArguments For The Existence Of God974 Words   |  4 PagesArguments for the existence of God through critical thinking and rationalization are called ontological, cosmological, teleological, or pragmeatic arguments. The most widely known of such arguments is that of St. Anselm from Proslogium of St. Anselm, which states that God is considered a perfect being unlike humans or any other world subject. The fact that he is perfect in a world of imperfection proves his existence. God is also the highest conceivable idea of perfection, and thereforeRead MoreThe Existence Of God : An Argument881 Words   |  4 PagesThe Existence of God The philosophical arguments presented in this document are not of religious text, nor scientific observation or established fact. Rather the premise of this God proof is bring together and share the various theories on which other God proofs have established foundations. I have heard it quoted that â€Å"Philosophy goes where hard science can t, or won t. Philosophers have a license to.† Therefore, with this in mind, I attest that it is more than problematic to construct anRead MoreArguments on the Existence of God602 Words   |  2 PagesGod’s existence may actually depend upon our belief in his existence but it is more plausible to believe that God exists using the different types of arguments such as the cosmological argument and ontological argument, Leibniz and the Principle of Sufficient Reason and the Problem of Evil, and the definition of basic belief as evidence. The Cosmological argument can be simplified into three reasons that everything that begins to exist has a cause; the universe began to exist, therefore the universeRead MoreArguments For The Existence Of God1137 Words   |  5 PagesArguments for the existence of God come in many different forms; some draw on history, some on science, some on personal experience, and some on philosophy. Descartes offered two arguments towards the existence of God: an informal proof in the third meditation and the ontological proof in the fifth meditation. Descartes believed that with the employment of a rational method of inquiry which applied some of the methods of analytic geometry to the study of philosophy, our ability to attain certaintyRead MoreThe Argument Of The Existence Of God1480 Words   |  6 PagesThe arguments trying to â€Å"prove† the existence of God are by far some of the most controversial philosophical arguments out there. When some of the people who created these philo sophies it was illegal or even punishable by death to even question his existence, let alone try to come up with a logical explanation to â€Å"prove† he is real. The two main arguments used today are the ontological argument and the cosmological argument. Neither one of these arguments are correct nor incorrect; moreover, theRead MoreThe Arguments For The Existence Of God940 Words   |  4 Pagesp. 209, question# 1 Among the numerous arguments for the existence of God, the argument of design stands as the most persuasive in terms of providing a logical basis for the absolute presence of God. This argument is concerned with the intricate nature of creation and existence: one must believe that there is a Supreme Being that designed the characteristics and features of every existing thing in the entire universe, both living and non-living. The precise and complicated design of the universeRead MoreThe Argument For The Existence Of God1411 Words   |  6 PagesMy paper scrutinizes numerous logical disputes for and alongside the presence of God. I shall argue that there’s no adequate evidence or inclusive arguments for the existence of God. It is grounded on the views of certain great philosophers and scientists of all of mankind. Generally speaking for myself, I would correspond to have faith that there is â€Å"God†. Regrettably, it’s awfully well-defined that the being built up on insightful faith is no longer a suitable custom to shadow. During the courseRead MoreThe Cosmological Argument For The Existence Of God Essay1556 Words   |  7 Pagesconcerning the existence of God. If God exists, we probably have to make him accountable. The universe would probably have a meaning and a purpose. Also, our very existence may not be cease after physical death. But if God does not exist, we are probably here by chance and we have no accountability to any transcendent. This life is probably all we have, so we should live as we please. The question arises - Does God exist? At first glance, it seems contradictory to prove the existence of something