DNA

Ubiquitin is an omnipresent protein found in all eukaryotes so far analysed. It is involved in several important processes, including protein turnover, chromosome structure and stress response. Parsley (Petroselinum crispum) contains at least two active polyubiquitin (ubi4) genes encoding hexameric precursor proteins. The deduced amino acid sequences of the ubiquitin monomers are identical to one another and to ubiquitin sequences from several other plant species. Analysis of the promoter region of one ubi4 gene revealed putative regulatory elements. In parsley plants, the ubi4 mRNAs were the predominant ubiquitin mRNAs and were present at comparable levels in all plant organs tested. In cultured parsley cells, high levels of ubiquitin gene expression remained unaffected by heat shock, elicitor or light treatment.

Originally prenatal diagnosis was confined to the diagnosis of metabolic disorders and depended on assaying enzyme levels in amniotic fluid. With the development of recombinant DNA technology, molecular diagnosis became possible for some genetic conditions late in the 1970s. Here we briefly review the history of molecular prenatal diagnostic testing, using Duchenne muscular dystrophy as an example, and describe how over the last 30 years we have moved from offering testing to a few affected individuals using techniques, such as Southern blotting to identify deletions, to more rapid and accurate PCR-based testing which identifies the precise change in dystrophin for a greater number of families. We discuss the potential for safer, earlier prenatal genetic diagnosis using cell free fetal DNA in maternal blood before concluding by speculating on how more recent techniques, such as next generation sequencing, might further impact on the potential for molecular prenatal testing. Progress is not without its challenges, and as cytogenetics and molecular genetics begin to unite into one, we foresee the main challenge will not be in identifying the genetic change, but rather in interpreting its significance, particularly in the prenatal setting where we frequently have no phenotype on which to base interpretation.

BACKGROUND. The diagnosis of malignant mesothelioma (MM) in serous effusions is difficult but may be achieved by the application of adjuvant methods. METHODS. The authors cytologically diagnosed 33 effusions as suspicious or positive for MM cells by using DNA-image cytometry (DNA-ICM), immunocytochemistry and AgNOR analysis. The authors further detected 9p21 deletions by chromosomal fluorescence in situ hybridization (FISH). In addition, 31 cases of metastatic carcinomas and 39 of tumor cell-negative effusions were investigated. All diagnoses were confirmed by histologic and/or clinical follow-up. RESULTS. DNA aneuploidy was found in 71% of MMs, 100% of metastatic carcinomas, and in none of the negative effusions. Calretinin was positive in 100% of MMs, in none of the metastatic carcinomas, and in 94.9% of negative effusions. BerEP4 showed positivity in 15.6% of MMs, 87.1% of metastatic carcinomas, and in none of the negative effusions. With AgNOR analysis, 89.3% of MMs and 96.7% of metastatic carcinomas showed 2.5 AgNOR dots as satellites and 4.5 as total AgNOR counts. 9p21 deletions were demonstrated in 90.9% of MM cases, 45.2% of metastatic carcinomas, and in none of the negative effusions. By cytology alone, 81.8% of MMs were identified unequivocally. Addition of DNA-ICM improved the prevalence of tumor cell detection to 87.9% and of AgNOR analysis to 97%. The introduction of 9p21 deletions by FISH improved this prevalence to 100%.

One of the best discriminators for the fertilization potential of human spermatozoa is sperm morphology. The problem in the assessment of the sperm morphological characteristics is their pleiomorphism. Examination of spermatozoa with the light microscope can provide only limited information on their internal structure. More detailed examination of sperm structure using electron microscopy can reveal major, often unsuspected ultrastructural abnormalities. Results and cut-off values for sperm analysis depend on the criteria for normal morphology. World Health Organization recommendations provide a classification suitable for clinical practice. Clinically reliable cut-off limits for normal sperm morphology according to strict Tygerberg criteria were suggested to be 4% in in-vitro fertilization procedures. Patients with severe sperm head abnormalities have a lower chance of establishing successful pregnancies, even though fertilization may be achieved. The outcome of intracytoplasmic sperm injection is not related to any of the standard semen parameters or to sperm morphology. Sperm decondensation defects and DNA anomalies may be underlying factors for the unrecognized derangements of the fertilizing capacity of spermatozoa, regardless of sperm morphology. Centrosome dysfunction may also represent a class of sperm defects that cannot be overcome simply by the insertion of a spermatozoon into the ooplasm. In this article an overview on the composition and ultrastructure of spermatozoa is presented, while emphasizing sperm ultrastructural and sperm DNA anomalies and their effects on fertilization.

T he acquired immunodeficiency syndrome (AIDS) emerged in the human population in the summer of 1981. According to the latest United Nations estimates, worldwide over 33 million people are infected with human immunodeficiency virus (HIV) and the prevalence rates continue to rise globally. To control the alarming spread of HIV, an urgent need exists for developing a safe and effective vaccine that prevents individuals from becoming infected or progressing to disease. To be effective, an HIV/AIDS vaccine should induce broad and long-lasting humoral and cellular immune responses, at both mucosal and systemic level. However, the nature of protective immune responses remains largely elusive and this represents one of the major roadblocks preventing the development of an effective vaccine. Here we summarize our present understanding of the factors responsible for resistance to infection or control of progression to disease in human and monkey that may be relevant to vaccine development and briefly review recent approaches which are currently being tested in clinical trials. Finally, the rationale and the current status of novel strategies based on nonstructural HIV-1 proteins, such as Tat, Nef and Rev, used alone or in combination with modified structural HIV-1 Env proteins are discussed. † These authors have contributed equally to this chapter.

Adnan Oktar (Harun Yahya)

İnsan, sadece bir kaç dakika için üzerinde yaşadığı dünyayı ve kendisine yaşam veren şeyleri dikkatlice düşündüğünde hayrete düşecektir. Devasa büyüklükte galaksiler barındıran bir boşlukta, uçsuz bucaksız büyüklükteki galaksilerden birinin içinde bulunan, yaşam için özel olarak var edilmiş bir gezegen üzerinde yaşamaktadır. Bu gezegen, yani Dünya, uçsuz bucaksız boşluğun içinde hiç durmadan dönmekte, evrendeki milyarlarca yıldızdan sadece biri olan Güneş yine aynı boşluk içinde yeryüzüne ışınlar yollamakta, bu ışınlar sayesinde Dünya ısınmakta, besin döngüsü, su döngüsü, azot döngüsü gerçekleşmekte, insan; hayvan, bitki ve mikroorganizmalarla birlikte kendisine sağlanan sayısız sebep vesilesiyle yaşayabilmektedir. Milyonlarca, milyarlarca detay bir araya getirilmiş, en güzel ve en kusursuz şekli ile insana sunulmuştur. Kimisi insanın yaşaması için gereken ihtiyaçları karşılarken, kimisi de bir güzellik, bir nimet olarak ona ikram edilmiştir. Bu detayların her biri bir sanattır, bir yaratılış harikasıdır. Bazen bu detaylar o kadar hayatidir ki, olmamaları durumunda yaşam durur, Dünya ölü bir gezegen halini alır. İnsanın ise, bunların çok büyük bir bölümünü değil yoktan var etmeye, benzerlerini bile yapmaya gücü yetmez.

Allah insana yaşamı boyunca nimetler sunar. İnsan, her an kopyalanan DNA'sı, her an aldığı nefes, her an atan kalbi, her nefeste soluduğu oksijen, her an dönen Dünya, her an hareket eden atomlar ve daha sayısız detay sayesinde yaşayabilmektedir. Çeşit çeşit yiyecekler, Dünya'ya "renk" getiren güneş ışınları, fotosentez yapan bitkiler, gökten inen su, bitkilere besin sunan mikroorganizmalar, denizler ve daha sayısız nimet bu dünyada yaşamın devamlılığına vesile olur. Dünya'nın konumu, Güneş'in, Ay'ın varlığı, bunların Dünya'ya uzaklıkları, Samanyolu galaksisi içindeki yerleri, büyüklükleri, eğimleri, yörüngeleri, içerdikleri tüm gazlar, moleküller, atomlar, insanın var olmasına uygun özel koşullarla yaratılmışlardır. İnsan, hayatta kalabilmesini sağlayacak sayısız nimetle birlikte kusursuz bir şekilde yaratıldığını düşünmelidir ve bunların tümü, kendisine önemli bir gerçeğin hatırlatıcılarındandır:

İnsanı yoktan yaratmış olan Allah'tır. Onu ve onun etrafını saran tüm güzellikleri, farkında olduğu veya olmadığı tüm nimetleri, bu nimetlerin en küçüğünü ve en büyüğünü sürekli olarak yaratan ve bunların her birinde hayranlık uyandırıcı detaylar var eden Yüce Allah'tır. Bu,  Allah'ın detay sanatıdır. Allah, sonsuz aklı ile insanların kavrayamadıkları, henüz detaylarını keşfedemedikleri sistemler yaratmış, her detayın içinde Kendi Yüceliğini ve kudretini gösteren daha da ince güzellikler var etmiştir. Yoktan var eden, her şeyi dilediği gibi takdir eden ve onları her an dilediği gibi yaratmaya kadir olan Yüce Allah için kuşkusuz bu son derece kolaydır. Allah dilerse, elbette bunların tümünü giderip yok edebilir. İnsana düşen, kendisine karşılıksız sunulan bu nimetlere şükretmek, Allah'a muhtaç olduğunu bilmek ve yalnızca O'na yönelmektir.

Adnan Oktar (Harun Yahya)

Today is the day of a major milestone in history. The materialist philosophy, once imposed on the mass of humanity under the guise of science, is ironically being defeated today by science itself.

Materialism, the philosophy which holds that everything is composed of matter and which denies the existence of God, is actually the contemporary version of paganism. Ancient pagans used to worship non-living beings like wooden or stone totem poles and considered them divine beings. Materialist philosophy, on the other hand, bases its claim on the belief that man and all other beings are created by atoms and molecules. According to this superstitious view, non-living atoms somehow organized themselves and over time acquired life and consciousness finally bringing man into being.

This superstitious belief of materialism is called "evolution". The belief in evolution, first introduced in the pagan cultures of the ancient Sumerians and then the ancient Greeks, was in a way revived in the 19th century by a group of materialist scientists and brought onto the world agenda. Charles Darwin is the best known of these scientists. The theory of evolution he advanced wasted the time of the world of science for 150 years, and despite its widely acknowledged flawed nature, has been hitherto sustained for purely ideological reasons.

However, as mentioned earlier, today, materialism is collapsing with a big boom. It is often stated that there were three important materialist theorists who steered the 19th century: Freud, Marx and Darwin. The theories of the first two were examined, tested, and, proving to be invalid, subsequently rejected in the 20th century. Nowadays, the theory of Darwin is also collapsing.

Non-syndromic hearing impairment (NSHI) is the most common form of deafness and presents with no other symptoms or sensory defects. Mutations in the gap junction gene GJB2 account for a high proportion of recessive NSHI. The GJB2 gene encodes connexin 26, which forms plasma membrane channels between cochlear cells. In Caucasian populations a single mutation, 35delG, accounts for most cases of NSHI. This mutation appears to be most prevalent in individuals of Mediterranean European descent, with carrier frequencies estimated as being as high as one in thirty. The 35delG region may be a mutational hotspot. The mutation arises from the deletion of a guanine from a six-guanine stretch and nearby microsatellite markers show little evidence for linkage disequilibrium. We believe that 35delG is an old mutation in a chromosomal region of high recombination. The genetic context of the 35delG mutation was examined to distinguish between an old or a recurring mutation. We identified two singlenucleotide polymorphisms (SNPs) immediately upstream of the first exon of GJB2. Polymerase chain reaction/restriction fragment length polymorphism analysis determined the SNP genotype of 35delG containing chromosomes from various populations, including Italy, Brazil, and North America. We found the same, relatively rare, polymorphism associated with the 35delG mutation in all populations studied. We have also examined microsatellite markers D13S175, which is 80 kb telomeric to GJB2, and D13S1316, which is 80 kb centromeric to GJB2. D13S175 appears to be in weak linkage disequilibrium with 35delG, while D13S1316 is less so. SNPs located between the 35delG mutation and the microsatellite markers show strong evidence of linkage disequilibrium. Taken together, these results indicate there has been substantial recombination near the 35delG mutation; however, we present evidence that the 35delG mutation arose in European and Middle Eastern populations from a single mutational event on a founder chromosome.

Temperature and pH are important parameters in food processing. Their effect on degradation of plasmid and plant DNA was investigated. Application of HPLC permitted us to quantitatively distinguish between covalently closed circular and nicked plasmid DNA. Nicking of plasmid DNA in buffer increased strongly at decreased pH (99% within 90 min at 65 °C/pH 4.0). A virtually identical degradation rate was observed in tomato serum. Temperature exerted only minor effects (15% within 90 min at 85 °C/pH 8.4). The combined effect of the parameters was additive. Degradation of plant DNA was monitored by determining the maximal detectable fragment length using polymerase chain reaction. With transgenic maize DNA in buffer, fragments up to 957 bp remained detectable for at least 90 min upon application of 65 °C at pH 4.0. Soymilk and tofu were produced from genetically modified soybeans. After boiling (10 min) of raw soymilk, the detectable fragment length decreased from 1,714 to 1,339 bp. The study of the final tofu product did not reveal any further DNA degradation. In commercial soy protein isolate, a rather highly processed food, DNA fragments of at least 714 bp were detected.

The ability to monitor the progress of single-molecule enzyme reactions is often limited by the need to use fluorogenic substrates. A method based on the principle of the Millikan oil drop experiment was developed to monitor the change in charge of substrates bound to a nanoparticle and offers a means of detecting single-enzyme reactions without fluorescence detection. As a proof of principle of the ability to monitor reactions that result in a change in substrate charge, polymerization on a single DNA template was detected. A custom oligonucleotide was synthesized that allowed for the attachment of single DNA templates to gold nanoparticles with a single polymer tether. The nanoparticles were then tethered to the surface of a microfluidic channel where the positions of the nanoparticles, subjected to an oscillating electric field, were monitored using dark field microscopy. With short averaging times, the signal-tonoise level was low enough to discriminate changes in charge of less than 1.2%. Polymerization of a long DNA template demonstrated the ability to use the system to monitor single-molecule enzymatic activity. Finally, nanoparticle surfaces were modified with thiolated moieties to reduce and/or shield the number of unproductive charges and allow for improved sensitivity.

In vitro sulfadoxine and pyrimethamine resistance has been associated with point mutations in the dihy- dropteroate synthase and dihydrofolate reductase domains, respectively, but the in vivo relevance of these point mutations has not been well established. To analyze the correlation between genotype and phenotype, 10 Cam- eroonian adult patients were treated with sulfadoxine-pyrimethamine and followed up for 28 days. After

Plasmid DNA (pDNA) used in vaccination and gene therapy has to be highly pure and homogenous, which point out necessity to develop efficient, reproducible and scalable downstream process. Convective Interaction Media (CIM) monolithic chromatographic supports being designed for purification of large molecules and nanoparticles seem to be a matrix of choice for pDNA purification. In present work we describe a pDNA purification process designed on two different CIM monolithic columns, based on anion-exchange (AEX) chromatography and hydrophobic interaction chromatography (HIC) chemistry. HIC monolith enabled separation of supercoiled (sc) pDNA from open circular (oc) pDNA, genomic DNA (gDNA) and endotoxins regardless to flow rates in the range at least up to 380 cm/h. Dynamic binding capacity of new HIC monolith is up to 4 mg of pDNA per milliliter of support. Combination of both chromatographic steps using optimized CaCl 2 precipitation enabled production of pure pDNA, satisfying all regulatory requirements. Process was found to be reproducible, scalable, and exhibits high productivity. In addition, in-line monitoring of pDNA purification process is shown, using CIM DEAE disk monolithic columns.

Site-specific insertion of 5-(3-aminopropyl)-2′-deoxyuridine (Z3dU) and 7-deaza-dG into the Dickerson-Drew dodecamers 5′-d((named DDD 2+Z10 )(X = Z3dU; Z = 7-deaza-dG) suggests a mechanism underlying the formation of interstrand N+2 DNA cross-links by nitrogen mustards, e.g., melphalan and mechlorethamine. Analysis of the DDD 2+Z10 duplex reveals that the tethered cations at base pairs A 5 · X 20 and X 8 · A 17 extend within the major groove in the 3′-direction, toward conserved Mg 2+ binding sites located adjacent to N+2 base pairs C 3 · Z 22 and Z 10 · C 15 . Bridging waters located between the tethered amines and either Z 10 or Z 22 O 6 stabilize the tethered cations and allow interactions with the N + 2 base pairs without DNA bending. Incorporation of 7-deaza-dG into the DDD 2+Z10 duplex weakens but does not eliminate electrostatic interactions between tethered amines and Z 10 O 6 and Z 22 O 6 . The results suggest a mechanism by which tethered N7-dG aziridinium ions, the active species involved in formation of interstrand 5′-GNC-3′ cross-links by nitrogen mustards, modify the electrostatics of ‡ Crystallographic coordinates and structure factors have been deposited in the Protein Data Bank as entries 2QEG (DDD Z10 ) and 2QEF (DDD 2+Z10 ).

Mouse embryonic stem (ES) cells were used as an experimental model to study the effects of electromagnetic fields (EMF). ES-derived nestin-positive neural progenitor cells were exposed to extremely low frequency EMF simulating power line magnetic fields at 50 Hz (ELF-EMF) and to radiofrequency EMF simulating the Global System for Mobile Communication (GSM) signals at 1.71 GHz (RF-EMF). Following EMF exposure, cells were analyzed for transcript levels of cell cycle regulatory, apoptosis-related, and neural-specific genes and proteins; changes in proliferation; apoptosis; and cytogenetic effects. Quantitative RT-PCR analysis revealed that ELF-EMF exposure to ES-derived neural cells significantly affected transcript levels of the apoptosis-related bcl-2, bax, and cell cycle regulatory "growth arrest DNA damage inducible" GADD45 genes, whereas mRNA levels of neural-specific genes were not affected. RF-EMF exposure of neural progenitor cells resulted in down-regulation of neural-specific Nurr1 and in up-regulation of bax and GADD45 mRNA levels. Short-term RF-EMF exposure for 6 h, but not for 48 h, resulted in a low and transient increase of DNA double-strand breaks. No effects of ELF-and RF-EMF on mitochondrial function, nuclear apoptosis, cell proliferation, and chromosomal alterations were observed. We may conclude that EMF exposure of ES-derived neural progenitor cells transiently affects the transcript level of genes related to apoptosis and cell cycle control. However, these responses are not associated with detectable changes of cell physiology, suggesting compensatory mechanisms at the translational and posttranslational level.

Circulating DNA is present in the blood of all individuals, but it has been found that cancer patients and patients with a variety of other conditions have increased amounts of these circulating DNA fragments in their blood. Even though more than 30 years of research have been done on this subject, the origin of these nucleic acid molecules is still not clear. We present evidence that does not support the general notion that apoptosis or necrosis is the major source of circulating free DNA. Active release of free circulating DNA by living cells may be a plausible mechanism. Disturbance of the equilibrium between the release of DNA by living cells and the mechanisms used for clearing this DNA may play the main role in the appearance of increased amounts of circulating DNA in the blood of individuals with different ailments. Elucidating the origin and the mechanism that cells use to release free circulating DNA into the blood may enhance the diagnostic and prognostic value of these nucleic acid molecules.

not seen. Using a lower estimate of the cluster mass, 2 £ 10 4 M ( (ref. 28), in stars sufficiently young for the present analysis, I estimate that the true deficit beyond 150 M ( in R136 is roughly four stars-that is, the result in the present work is more statistically significant by this measure. If the deficit of massive stars in R136 is real, then it represents another measurement of the upper mass cut-off.

In this paper, we propose a new methodology for analysis of microarray images. First, a new gridding algorithm is proposed for determining the individual spots and their borders. Then, a Gaussian mixture model (GMM) approach is presented for the analysis of the individual spot images. The main advantages of the proposed methodology are modeling flexibility and adaptability to the data, which are well-known strengths of GMM. The maximum likelihood and maximum a posteriori approaches are used to estimate the GMM parameters via the expectation maximization algorithm. The proposed approach has the ability to detect and compensate for artifacts that might occur in microarray images. This is accomplished by a model-based criterion that selects the number of the mixture components. We present numerical experiments with artificial and real data where we compare the proposed approach with previous ones and existing software tools for microarray image analysis and demonstrate its advantages.

The early evolution of the major groups of derived non-avialan theropods is still not well understood, mainly because of their poor fossil record in the Jurassic. A well-known result of this problem is the 'temporal paradox' argument that is sometimes made against the theropod hypothesis of avian origins 1 . Here we report on an exceptionally well-preserved small theropod specimen collected from the earliest Late Jurassic Tiaojishan Formation of western Liaoning, China 2 . The specimen is referable to the Troodontidae, which are among the theropods most closely related to birds. This new find refutes the 'temporal paradox' 1 and provides significant information on the temporal framework of theropod divergence. Furthermore, the extensive feathering of this specimen, particularly the attachment of long pennaceous feathers to the pes, sheds new light on the early evolution of feathers and demonstrates the complex distribution of skeletal and integumentary features close to the dinosaur-bird transition.

The structural and spectroscopic properties of [Ru(phen) 2 (dppz)] 2+ and [Ru(tap) 2 (dppz)] 2+ (phen = 1,10phenanthroline; tap= 1,4,5,8-tetraazaphenanthrene; dppz = dipyridophenazine ) have been investigated by means of density functional theory (DFT), time-dependent DFT (TD-DFT) within the polarized continuum model (IEF-PCM) and quantum mechanics/molecular mechanics (QM/MM) calculations. The model of the Δ and Λ enantiomers of Ru(II) intercalated in DNA in the minor and major grooves is limited to the metal complexes intercalated in two guanine-cytosine base pairs. The main experimental spectral features of these complexes reported in DNA or synthetic polynucleotides are better reproduced by the theoretical absorption spectra of the Δ enantiomers regardless of intercalation mode (major or minor groove). This is especially true for [Ru(phen) 2 (dppz)] 2+ . The visible absorption of [Ru(tap) 2 (dppz)] 2+ is governed by the MLCT tap transitions regardless of the environment (water, acetonitrile or bases pair), the visible absorption of [Ru(phen) 2 (dppz)] 2+ is characterized by transitions to metal-to-ligand-charge-transfer MLCT dppz in water and acetonitrile and to MLCT phen when intercalated in DNA. The response of the IL dppz state to the environment is very sensitive. In vacuum, water and acetonitrile these transitions are characterized by significant oscillator strengths and their positions depend significantly on the medium with blue shifts of about 80 nm when going from vacuum to solvent. When the complex is intercalated in the guanine-cytosine base pairs the 1 IL dppz transition contributes mainly to the band at 370 nm observed in the spectrum of [Ru(phen) 2 (dppz)] 2+ and to the band at 362 nm observed in the spectrum of [Ru(tap) 2 (dppz)] 2+ .

Genitopatellar syndrome (GPS) and Say-Barber-Biesecker-Young-Simpson syndrome (SBBYSS or Ohdo syndrome) have both recently been shown to be caused by distinct mutations in the histone acetyltransferase KAT6B (a.k.a. MYST4/MORF). All variants are de novo dominant mutations that lead to protein truncation. Mutations leading to GPS occur in the proximal portion of the last exon and lead to the expression of a protein without an activation domain. Mutations leading to SBBYSS occur either throughout the gene, leading to nonsense-mediated decay, or more distally in the last exon. Features present only in GPS are contractures, anomalies of the spine, ribs and pelvis, renal cysts, hydronephrosis and agenesis of the corpus callosum. Features present only in SBBYSS include long thumbs and long great toes and lacrimal duct abnormalities. Several features occur in both, such as intellectual disability, congenital heart defects, genital and patellar anomalies. We propose that haploinsufficiency or loss of a function mediated by the Cterminal domain causes the common features, whereas gain-of-function activities would explain the features unique to GPS. Further molecular studies and the compilation of mutations in a database for genotype-phenotype correlations (www.LOVD.nl/KAT6B) might help tease out answers to these questions and understand the developmental programs dysregulated by the different truncations.

Ancient DNA research from its inception in the 1980s is reviewed in the context of pre-Columbian archaeology. Changing approaches to the recovery of DNA from human remains are followed, and the accumulated records considered in the light of hypotheses about the post-Columbian depletion and augmentation of the New World gene pool.

A new method for biolistic delivery of nucleic acids using a combination of cationic micro-and nanoparticles is reported. The new method is simpler to perform than the conventional calcium/ spermidine-based formulations and shows eleven-fold improved nucleic acid binding capacity and dose-dependent performance both for in vitro and in vivo applications relative to either the conventional preparation or our recently reported direct cationic microparticle method. These features may enable higher throughput gene delivery and genetic immunization programs and open new venues for biolistic delivery method.

38 sc ientific american mind augu st /s eptember 20 0 8 a n tO n i O m . rO s a r i O G et t y I mage s ( no se) ; G e t t Y i m aG e s ( un smiling c ouple) ; J U Pit e rimaGe s ( armpit and smiling c ouple) ; i s t O c K P H O t O : J O e m c da n i e L ( dir t y sock s ) , Ja KU B s e m e n i U K ( lemon s ) , c Hri s t ine GL ade (urine s ample) , OK taY Or taKc iO GLU

Context: Primary adrenal insufficiency (PAI) is a life-threatening condition that is often due to monogenic causes in children. Although congenital adrenal hyperplasia (CAH) occurs commonly, several other important molecular causes have been reported, often with overlapping clinical and biochemical features. The relative prevalence of these conditions is not known, but making a specific diagnosis can have important implications for management.

Causes of late Quaternary extinctions of large mammals (''megafauna'') continue to be debated, especially for continental losses, because spatial and temporal patterns of extinction are poorly known. Accurate latest appearance dates (LADs) for such taxa are critical for interpreting the process of extinction. The extinction of woolly mammoth and horse in northwestern North America is currently placed at 15,000 -13,000 calendar years before present (yr BP), based on LADs from dating surveys of macrofossils (bones and teeth). Advantages of using macrofossils to estimate when a species became extinct are offset, however, by the improbability of finding and dating the remains of the last-surviving members of populations that were restricted in numbers or confined to refugia. Here we report an alternative approach to detect 'ghost ranges' of dwindling populations, based on recovery of ancient DNA from perennially frozen and securely dated sediments (sedaDNA). In such contexts, sedaDNA can reveal the molecular presence of species that appear absent in the macrofossil record. We show that woolly mammoth and horse persisted in interior Alaska until at least 10,500 yr BP, several thousands of years later than indicated from macrofossil surveys. These results contradict claims that Holocene survival of mammoths in Beringia was restricted to ecologically isolated high-latitude islands. More importantly, our finding that mammoth and horse overlapped with humans for several millennia in the region where people initially entered the Americas challenges theories that megafaunal extinction occurred within centuries of human arrival or were due to an extraterrestrial impact in the late Pleistocene. extinction ͉ permafrost ͉ megafauna ͉ Beringia

A 22-year-old captive Brazilian rainbow boa (Epicrates cenchria cenchria) gave birth to four offspring after being housed with a vasectomized male. Sexual reproduction as a result of failed prior vasectomy, recanalization of the vas deferens, or prolonged sperm storage was ruled out using the clinical history, histopathology, and gross necropsy. Short tandem repeat (STR) DNA markers were genotyped in the male, female, and four offspring. None of the offspring possessed a diagnostic STR allele present in the potential sire. In addition, all offspring were homozygous at each STR locus evaluated, supporting parthenogenetic reproduction. This is the first report of parthenogenesis in a Brazilian rainbow boa and has implications for the conservation of reptiles maintained in captive breeding programs.

Introduction B cells have many different roles in systemic lupus erythematosus (SLE), ranging from autoantigen recognition and processing to effector functions (for example, autoantibody and cytokine secretion). Recent studies have shown that intracellular nucleic acid-sensing receptors, Toll-like receptor (TLR) 7 and TLR9, play an important role in the pathogenesis of SLE. Dual engagement of rheumatoid factor-specific AM14 B cells through the B-cell receptor (BCR) and TLR7/9 results in marked proliferation of autoimmune B cells. Thus, strategies to preferentially block innate activation through TLRs in autoimmune B cells may be preferred over non-selective B-cell depletion.

In a healthy body, ROS (reactive oxygen species) and antioxidants remain in balance. When the balance is disrupted towards an overabundance of ROS, oxidative stress (OS) occurs. OS results from an imbalance between prooxidants (free radical species) and the body's scavenging ability (antioxidants). ROS are a double-edged swordthey serve as key signal molecules in physiological processes but also have a role in pathological processes. The production of ROS is a normal physiological event in various organs including the testis. Overproduction of ROS can be detrimental to sperm and being associated with male infertilities. The excessive generation of ROS by abnormal spermatozoa, contaminating leukocytes and by a various type of pollutants has been identified as detrimental etiologies for male infertilities Free radicals are substances with one or more unpaired electrons, which are formed as a results of many physiological and pathological cellular metabolic processes, especially in mitochondria. Enzymatic (Catalase, superoxide dismutase) and non enzymatic (vitamins A and E) natural antioxidant defense mechanisms exist; however, these mechanisms may be overcome, causing lipid peroxidation to take place. For example, breakdown in the cells results in the formation of molecules whose further metabolism in the cell leads to ROS production. Thus increased OS stimulates the activity of enzymes called cytochrome P450, which contribute to ROS production. . Oxidative stress index (OSI) was calculated as ([TOS/TAS] x 100). TOS and OSI were significantly higher and PON-1 activity and TAS were significantly lower in subfertile male with abnormal semen parameters than in male with idiopathic subfertility and fertile donors. PON-1 activity was also strongly correlated with sperm concentration, motility, and morphology in the overall group. The receiver operating characteristic curve analysis revealed a high diagnostic value for PON-1 activity with respect to male-factor sub fertility. ROS may cause infertility by two principal mechanisms, first ROS damage the sperm membrane which in turn reduces the sperm motility and ability to fuse with the oocyte secondly, and ROS directly damage sperm DNA, compromising the paternal genomic contribution to the embryo. Oxidative stress due to excessive production of ROS, impaired antioxidant defense mechanisms, or both precipitates a range of pathologies that are currently believed to negatively affect the male reproductive function. Oxidative stress-induced damage to sperm may be mediated by lipid peroxidation of the sperm plasma membrane, reduction of sperm motility, and damage to the DNA in the sperm nucleus, as the production of ROS is one of the principal mechanisms by which neutrophils destroy pathogens, it is not surprising that seminal leukocytes have the potential to cause oxidative stress. Despite the established role of OS in the pathogenesis of male infertility, there is a lack of consensus as to the clinical utility of seminal OS testing in an infertility clinic. One important reason for the inability to utilize the OS test in clinical practice is related to the lack of a standard protocol for assessment of seminal OS. Antioxidants are powerful and there are few trials investigating antioxidant supplementation in male reproduction. Several researches indicate that the diagnostic and prognostic capabilities of the seminal OS test are beyond those of conventional tests of sperm quality and function. The OS test can accurately discriminate between fertile and infertile male and identify male with a clinical diagnosis of male-factor infertility that are likely to initiate a pregnancy when followed over a period of time. We strongly believe that incorporating such a test into the routine andrology workup is an important step for the future of the male infertility practice. The resulting state of the cell, known as (OS) can lead to cell injury. ROS production and Lipid peroxidation, free radical and oxidative stress in relation to fertility are the aim of this review [Magda M El-Tohamy. The mechanisms by which Oxidative Stress and Free Radical Damage produces Male infertility.

Humans have been doing genetic engineering, a technology which is transforming our world, for thousands of years on a wide range of plants, animals and micro organism and have applications in the field of medicine, research, industry and agriculture. The rapid developments in the field of genetic engineering have given a new impetus to biotechnology. This introduces the possibility of tailoring organisms in order to optimize the production of established or novel metabolites of commercial importance and of transferring genetic material from one organism to another. In order to achieve potential benefits of genetic engineering the only need is to develop perfect tools and techniques. Once it has been perfected then all of the problems associated with food production can be solved, the world environment can be restored, and human health and lifestyle will improve beyond imagination. No doubt that there are almost no limits to what can be achieved through responsible genetic engineering. Classical field of genetic engineering and some of its advancements are discussed in this review.

The rapid identification and characterization of genes of neurological relevance holds great potential for offering insight into the diagnosis, management, and understanding of the pathophysiologic mechanisms of neurological diseases. This volume in the Methods in Molecular Biology™ series was conceived to highlight many of the contemporary methodological approaches utilized for the characterization of neurologically relevant gene mutations and their protein products. Although an emphasis has been placed upon descriptions of methodologies with a defined clinical utility, it is hoped that Neurogenetics: Methods and Protocols will appeal not only to clinical laboratory diagnosticians, but also to clinicians, and to biomedical researchers with an interest in advances in disease diagnosis and the functional consequences of neurologically relevant gene mutations. To meet this challenge, more than 60 authors graciously accepted my invitation to contribute to the 32 chapters of this book. Through their collective commitment and diligence, what has emerged is a comprehensive and timely treatise that covers many methodological aspects of mutation detection and screening, including discussions on quantitative PCR, trinucleotide repeat detection, sequence-based mutation detection, molecular detection of imprinted genes, fluorescence in situ hybridization (FISH), in vitro protein expression systems, and studies of protein expression and function. I would like to take this opportunity to formally thank my colleagues for their effort and dedication to this work. This book would not have been possible without the guidance and wisdom of the Series Editor, Professor John M. Walker, whose intimate knowledge of the nuances of the editorial process made my job somewhat less intimidating. I would also like to thank Thomas Lanigan, President of Humana Press, who enthusiastically embraced the book concept and my original prospectus from the very beginning, and Craig Adams, also at Humana Press, for transforming the individual chapters into their final form.

Adnan Oktar (Harun Yahya)

İlk ortaya atıldığı günden bu yana bazı çevrelerce ideolojik bir zorunluluk olarak ilgi gören evrim teorisi, günümüzde de aynı zorunluluk sebebiyle yoğun bir propaganda ile savunulmaktadır. Kimi bilimsel yayınlarda, okullarda ve bazı medya organlarında canlılığın kökeni konusunda ispatlanmış bir gerçek gibi sunulmaktadır. Evrim teorisi bilim adamlarının bir kısmı tarafından da büyük bir bağlılıkla savunulduğu için, insanların birçoğu bu teoriyi bilimsel açıdan geçerli zannetmektedirler.

Oysa evrim teorisi, 150 yıldır paleontoloji, genetik, mikrobiyoloji, biyokimya, biyofizik gibi bilim dalları tarafından çürütülmüştür! Bu bilim dalları tarafından ortaya konan sayısız bulgu evrimin hiçbir zaman gerçekleşmediğini çok açık ve kesin olarak göstermektedir.

Bu yazıyı okuyan birçok insanın aklına doğal olarak şöyle bir düşünce gelebilir: "Evrim teorisini savunanlar bilim adamlarıdır. Evrim teorisinin geçersiz olduğunu ispatlayan delilleri bulanlar da yine bilim adamlarıdır. Bu durumda bilim adamları herhalde iki gruba bölünmüştür; bir kısmı evrimi savunurken, diğerleri evrimi çürüten delilleri bulmuşlardır."

Oysa bu düşünce tam olarak doğruyu ifade etmemektedir. Çünkü evrim teorisini savunanlar, evrimin gerçekleştiğine dair hiçbir delil bulamazlarken, bizzat kendileri evrim teorisini çürüten delilleri yine kendi elleriyle bulmaktadırlar.

Kuşkusuz bu son derece çelişkili ve samimiyetsiz bir durumdur. Bu insanların, hem ellerindeki teoriyi çürütecek delilleri bulurken, hem de o teoriyi ısrarla savunmaya çalışmaları son derece anlamsızdır. Ancak gerçek işte tam da böyledir.