DNase Research Articles

DNase X Is a Glycosylphosphatidylinositol-anchored Membrane Enzyme That Provides a Barrier to Endocytosis-mediated Transfer of a Foreign Gene

DNase X is the first mammalian DNase to be isolated that is homologous to DNase I. In this study, we have examined its function using a novel monoclonal antibody and showed it to be expressed on the cell surface as a glycosylphosphatidylinositolanchored membrane protein. High level expression was observed in human muscular tissues and in myotubes obtained in vitro from RD rhabdomyosarcoma cells. We observed that RD myotubes incorporated a foreign gene, lacZ, by endocytosis but that expression of the encoded coding product, beta-galactosidase, was strongly inhibited. Overexpression of DNase X inhibited endocytosis-mediated gene transfer, whereas knockdown of DNase X with small interfering RNA had the opposite effect. These results reveal that DNase X provides a cell surface barrier to endocytosis-mediated gene transfer.

Degradation of nuclear DNA by DNase II‐like acid DNase in cortical fiber cells of mouse eye lens

The eye lens is composed of fiber cells that differentiate from epithelial cells on its anterior surface. In concert with this differentiation, a set of proteins essential for lens function is synthesized, and the cellular organelles are degraded. DNase II-like acid DNase, also called DNase IIbeta, is specifically expressed in the lens, and degrades the DNA in the lens fiber cells. Here we report that DNase II-like acid DNase is synthesized as a precursor with a signal sequence, and is localized to lysosomes. DNase II-like acid DNase mRNA was found in cortical fiber cells but not epithelial cells, indicating that its expression is induced during the differentiation of epithelial cells into fiber cells. Immunohistochemical and immunocytochemical analyses indicated that DNase II-like acid DNase was colocalized with Lamp-1 in the lysosomes of fiber cells in a relatively narrow region bordering the organelle-free zone, and was often found in degenerating nuclei. A comparison by microarray analysis of the gene expression profiles between epithelial and cortical fiber cells of young mouse lens indicated that some genes for lysosomal enzymes (cathepsins and lipases) were strongly expressed in the fiber cells. These results suggest that the lysosomal system plays a role in the degradation of cellular organelles during lens cell differentiation.

Application of different test systems and biochemical indicators for environmental monitoring of the Troitsa Bay, Sea of Japan

The DNase express test system and the SF-bioassay (sea urchin sperm fertilization bioassay) were used to assess the quality of sea and bottom sediment water sampled at 8 stations of the Troitsa Bay (Possyet Bay, Sea of Japan). The specific activities of 8 enzymes (alkaline and acid DNases, RNases, phosphatases and phosphodiesterases) were determined for the liver and gill tissues of the mussel Crenomytilus grayanus sampled at the same stations. The activities of all enzymes were higher in the liver than in the gills, and the activity of acid nucleases in both tissues was significantly higher than that of alkaline nucleases. The specific activities of some nucleases in the mussel tissues correlated to the level of seawater pollution. The activities of acid RNases and phosphodiesterases decreased in both tissues upon an increase in water pollution, up to 1.5–3 times in the gills, while the activities of alkaline and acid phosphatases and phosphodiesterases increased 1.5–4 times. Thus, the specific activities of acid RNases, phosphatases and phosphodiesterases from mussel liver can be a useful indicator of marine pollution. This approach can be used for biotesting of ocean water and for assessment of the biological condition of invertebrates adapting to various ecological and anthropogenic effects.

The modulator effect of GH on skeletal muscle lysosomal enzymes is dietary protein dependent

Objective The purpose of this work is to determine whether changes in dietary protein level could alter the modulator effect that GH has on the muscle lysosomal system by influencing the hydrolytic activities of cathepsin D, acid RNase and DNase II and the participation of these enzymes in muscle growth. Design BALB/c female mice were fed a diet containing 20% (HP) or 12% (MP) protein ad libitum and were treated with either saline (s) or rhGH (GH) (74 ng/g) for 29 days. Body weight and feed intake were recorded daily. At 25, 30, 35, 40, 45 and 50 days of age, five mice from each group were slaughtered and nucleic acids and protein concentrations and cathepsin D, acid RNase and DNase II activities in gastrocnemius muscle were analysed. Correlation coefficients were used to analyse the links between the activity of each enzyme with its substrate. Results GH-treatment induced a depletion–recovery response in muscle growth through a compensatory mechanism. Changes in protein content, DNA and RNA concentrations were related to changes in lysosomal enzyme activities. Muscle cathepsin D activity in saline mice fell as the dietary protein concentration increased. GH-treatment reversed this effect by enhancing the proteolytic activity in muscle of well-fed mice and inhibiting it in mice fed a 12% protein diet. This inversion appears to be related to the different mechanism elicited by GH-treatment on skeletal muscle protein growth in each dietary group. An opposite trend was observed in muscle acid nuclease activities. Acid RNase and DNase II increased according to the dietary protein concentration, since a 12% protein diet induced a lower catabolism, especially on muscle DNA of saline mice. In contrast, GH-treatment decreased acid RNase and DNase II activities, but only in mice fed a 20% protein diet, perhaps leading to spare muscle RNA for protein synthesis, as well as to the inhibition of DNA degradation during catch-up growth. A lower dietary protein concentration appeared to reverse the GH protective effect on nucleic acids. Conclusions GH seems to act as a dietary protein-dependent modulator of the skeletal muscle lysosomal enzyme activity. These lysosomal enzymes play a role during muscle growth in GH-treated post-weaning mice by modifying muscle protein and DNA and RNA degradation.

Purification of monocytes from cryopreserved mobilized apheresis products by elutriation with the Elutra® device

The Elutra® biomedical device allows semi-automatic enrichment of monocytes by elutriation, using a single-use, closed and cGMP compliant tubing set, in a cost effective way. The procedure has been validated using fresh apheresis products from nonmobilized donors. We here evaluated the possibility of using Elutra® to enrich monocytes from frozen/thawed apheresis products collected from mobilized healthy donors. Frozen apheresis products from 6 G CSF mobilized donors were thawed and used in 16 elutriation procedures. We compared the recovery and purity of enriched monocytes using different buffer compositions and elutriation profiles. Elutriated monocytes were cultured to generate mature dendritic cells (DCs). Depending in part of the initial granulocyte contamination in the apheresis product, the use of Desoxyribo Nuclease (DNAse) to avoid aggregation, was needed through only the initial steps or throughout the elutriation process. The average monocyte recovery was 85 ± 31%. The average purity was 73 ± 9%. The recovery of mature DC at d8 of culture was 20 ± 6% of the input monocyte numbers. We conclude that Elutra® allows the purification of monocytes from thawed mobilized apheresis. It requires no pre-processing of the cell product before elutriation, and allows the generation of phenotypically mature DC in quantities that are compatible with a clinical use.

Deoxyribonuclease 1 aggravates acetaminophen-induced liver necrosis in male CD-1 mice

An overdose of acetaminophen (APAP) (N-acetyl-p-aminophenol) leads to hepatocellular necrosis induced by its metabolite N-acetyl-p-benzoquinone-imine, which is generated during the metabolic phase of liver intoxication. It has been reported that DNA damage occurs during the toxic phase; however, the nucleases responsible for this effect are unknown. In this study, we analyzed the participation of the hepatic endonuclease deoxyribonuclease 1 (DNASE1) during APAP-induced hepatotoxicity by employing a Dnase1 knockout (KO) mouse model. Male CD-1 Dnase1 wild-type (WT) (Dnase1+/+) and KO (Dnase1-/-) mice were treated with 2 different doses of APAP. Hepatic histopathology was performed, and biochemical parameters for APAP metabolism and necrosis were investigated, including depletion of glutathione/glutathione-disulfide (GSH+GSSG), beta-nicotinamide adenine dinucleotide (NADH+NAD+), and adenosine triphosphate (ATP); release of aminotransferases and Dnase1; and occurrence of DNA fragmentation. As expected, an APAP overdose in WT mice led to massive hepatocellular necrosis characterized by the release of aminotransferases and depletion of hepatocellular GSH+GSSG, NADH+NAD+, and ATP. These metabolic events were accompanied by extensive DNA degradation. In contrast, Dnase1 KO mice were considerably less affected. In conclusion, whereas the innermost pericentral hepatocytes of both mouse strains underwent necrosis to the same extent independent of DNA damage, the progression of necrosis to more outwardly located cells was dependent on DNA damage and only occurred in WT mice. Dnase1 aggravates APAP-induced liver necrosis.

Involvement of DNase II-Like Acid DNase in the Cataract Formation of the UPL Rat and the Shumiya Cataract Rat

The loss of organelles and DNA is important to ensure transparency of the lenses, and DNase II-like acid DNase (also called DNase IIbeta, DLAD) is related to the loss of organelles and DNA in the lenses. We investigated the relation between the degradation of DNA and DLAD mRNA expression in the lenses of two hereditary cataract rats, the UPL rat (UPLR) and the Shumiya cataract rat (SCR), during cataract development. Undigested DNA was detected in the lens cortexes of normal UPLRs and SCRs, and undigested DNA was degraded in the lens nuclei of normal UPLRs and SCRs. DLAD does not affect common cataract formation, since DLAD mRNA expression levels in the lenses of cataractous SCRs were not changed with an increase in age, and undigested DNA was degraded in the lens nuclei of cataractous SCRs. On the other hand, an accumulation of undigested DNA was found in the lens nuclei of cataractous UPLRs at 46 and 53 d of age with opaque lenses, and the decrease in DLAD mRNA expression levels occurred prior to the accumulation of undigested DNA in the lens nuclei. It is possible that UPLRs are a good model for cataract caused by a decrease of DNA degradation in the lenses.

Severe Defects in Proliferation and Differentiation of Lens Cells in Foxe3 Null Mice

During mouse eye development, the correct formation of the lens occurs as a result of reciprocal interactions between the neuroectoderm that forms the retina and surface ectoderm that forms the lens. Although many transcription factors required for early lens development have been identified, the mechanism and genetic interactions mediated by them remain poorly understood. Foxe3 encodes a winged helix-forkhead transcription factor that is initially expressed in the developing brain and in the lens placode and later restricted exclusively to the anterior lens epithelium. Here, we show that targeted disruption of Foxe3 results in abnormal development of the eye. Cells of the anterior lens epithelium show a decreased rate of proliferation, resulting in a smaller than normal lens. The anterior lens epithelium does not properly separate from the cornea and frequently forms an unusual, multilayered tissue. Because of the abnormal differentiation, lens fiber cells do not form properly, and the morphogenesis of the lens is greatly affected. The abnormally differentiated lens cells remain irregular in shape, and the lens becomes vacuolated. The defects in lens development correlate with changes in the expression of growth and differentiation factor genes, including DNase II-like acid DNase, Prox1, p57, and PDGFalpha receptor. As a result of abnormal lens development, the cornea and the retina are also affected. While Foxe3 is also expressed in a distinct region of the embryonic brain, we have not observed abnormal development of the brain in Foxe3(-/-) animals.

Sequence-dependent cleavage of HBV DNA by combining with triple helix-forming oligodeoxyribonucleotides modified with manganese porphyrin in vitro

To observe the ability of triple helix-forming oligonucleotides (TFO) modified with manganese porphyrin to combine with and cleave HBV DNA fractions. The ends of TFO were modified with manganese porphyrin and acridine; At 37 degrees C and pH 7.4 condition in vitro, TFO modified with manganese porphyrin and acridine were bound with 32P labeled HBV DNA fragments, the affinity and specificity binding to target sequence were tested by electrophoretic mobility shift and DNase 1 footprinting assays, the ability to cleave HBV DNA fractions was observed with cleavage experiments. TFO modified with manganese porphyrin and acridine could bind to target sequence in a sequence-dependent manner with Kd values of 3.5 x 10(-7) mol/L and a relative affinity of 0.008. In the presence of KHSO5, TFO modified with manganese porphyrin and acridine could cleave target sequence in the region forming triple DNA. In the presence of KHSO5, TFO modified with manganese porphyrin and acridine could cleave target HBV-DNA in sequence-dependent manner.

Picrotoxin-induced convulsions and lysosomal function in the rat brain

The effect of picrotoxin-induced convulsions on lysosomal function in rat brain were evaluated by measuring the free as well as total acid phosphatase, cathepsin D, acid ribonuclease (RNAse II) and acid deoxyribonuclease (DNAse II) activities. Following picrotoxin treatment the free RNAse II activity increased whereas the total activities of practically all the other enzymes decreased. Paradoxically, the cathepsin D activity, free as well the total was completely abolished. In case of all the enzymes the ratio of Total activity/Free activity decreased indicating increased lysosomal membrane fragility which could lead to process of neurodegeneration in the epileptic animals.

Effect of antimalarials treatment on rat liver lysosomal function—Anin vivo study

Effects of treatmentin vivo with the antimalarials:chloroquine (CQ), primaquine (PQ) and quinine(Q) on lysosomal enzymes and lysosomal membrane integrity were examined. Treatment with the three antimalarials showed an apparent increase in the membrane stability. CQ treatment resulted in increase in both the 'free' and 'total' activities of all the enzymes i.e. acid phosphatase, RNase II, DNase II and cathepsin D. PQ treatment lowered the 'free' and 'total' activities of acid phosphatase and cathepsin D, but the DNase II activities increased. Treatment with Q resulted in increased 'free' and 'total' activities of RNase II and DNase II. While 'free' activities of acid phosphatase and cathepsin D were low; the 'total' activities increased significantly. Our results suggest that a generalized increase in free nucleases activities following prolonged treatment with antimalarials may lead to cell damage and/or necrosis.

Intestinal DNases of 36 and 38.5 kDa from the parasitic nematode Haemonchus contortus have non-classic DNase characteristics and produce DNA fragments with 3′-hydroxyls

Treatment with the anthelmintic fenbendazole induces fragmentation of genomic DNA in intestinal cells of Haemonchus contortus. This effect is characterized by DNA fragments with 3′-hydroxyls (OH). Investigation into DNases responsible identified intestinal DNase activities that produce DNA fragments with 3′-OH. However, this interpretation was complicated by a mixture of activities in the intestinal fractions evaluated. In addition, intestinal activities displayed non-classic characteristics. Here it is shown that heparin sulfate (HS) fractionation enriched for intestinal DNases that produce 3′-OH. The 2.0 M NaCl fraction of HS contained DNase activity that produced 3′-OH with minimal contamination by activity that produced 3′-phosphates (P). 3′-OH were produced under acidic (pH 5.0) or neutral (pH 7.0) conditions by DNases in this fraction. These DNases were sensitive to EDTA under each condition. Furthermore, EDTA-sensitive DNase activity in this fraction digested H. contortus intestinal cell nuclear DNA in histological sections, producing 3′-OH under acidic and neutral conditions. DNases at 36 and 38.5 kDa in this fraction each produced 3′-OH at pH 5.0 when gel eluted, and each activity was sensitive to EDTA. Hence, the 36 and 38.5 kDa DNases in the 2.0 M NaCl HS intestinal fraction have characteristics expected for candidate DNases that mediate DF in H. contortus intestinal cell nuclei induced by fenbendazole. DNase activity that produces 3′-OH under acidic condition with sensitivity to EDTA is unconventional for classic acidic or neutral DNases and is a unique finding for nematodes. Excretory/secretory products from the worm and whole worm lysates were also explored as sources to fractionate intestinal DNases identified. HS fractionation of those worm samples did not clearly resolve the intestinal DNases of interest, although DNases with distinct characteristics were identified in each source.

Purification and characterization of an acid deoxyribonuclease from the cultured mycelia of Cordyceps sinensis.

A new acid deoxyribonuclease (DNase) was purified from the cultured mycelia of Cordyceps sinensis, and designated CSDNase. CSDNase was purified by (NH(4))(2)SO(4) precipitation, Sephacryl S-100 HR gel filtration, weak anion-exchange HPLC, and gel filtration HPLC. The protein was single-chained, with an apparent molecular mass of ca. 34 kDa, as revealed by SDS-PAGE, and an isoelectric point of 7.05, as estimated by isoelectric focusing. CSDNase acted on both double-stranded (ds) and single- stranded (ss) DNA, but preferentially on dsDNA. The optimum pH of CSDNase was pH 5.5 and its optimum temperature 55. The activity of CSDNase was not dependent on divalent cations, but its enzymic activity was inhibited by high concentration of the cation: MgCl(2) above 150 mM, MnCl(2) above 200 mM, ZnCl(2) above 150 mM, CaCl(2) above 200 mM, NaCl above 300 mM, and KCl above 300 mM. CSDNase was found to hydrolyze DNA, and to generate 3-phosphate and 5-OH termini. These results indicate that the nucleolytic properties of CSDNase are essentially the same as those of other well-characterized acid DNases, and that CSDNase is a member of the acid DNase family. To our knowledge, this is the first report of an acid DNase in a fungus.

474. The Barrier to Liposomal Transfection Is Enhanced by DNASES Lacking Lysosomal-Targetting Glycosylation

Prior studies suggested that liposome-transfected plasmids, to achieve expression, had to evade post-endocytic lysosomal degradation by Dnase 1 (D1) and/or Dnase II; however, we have shown that the main circulating nuclease barrier to liposomal gene transfection is almost certainly the macrophage-secreted enzyme Dnase1-like 3 (Dnase1l3 or D3) [Wilber et al. Mol Ther 2002]. In addition, we found this barrier is not cell-autonomous, but is conveyed upon HeLa cells by media conditioned either by cells expressing transfected D3 or interferon-gamma induced macrophages. By immunoprecipitation, western, and zymogram, the only Dnase secreted by induced macrophages is the 28 kDa D3 enzyme. This innate immunity function is postulated to exist to protect cells against the entry and expression of potentially mutagenic or infectious extracellular DNA. While homologous to D1 over its core nuclease sequence, D3 has a distinct basic C-terminus. If this C-terminus is truncated from D3, the remaining protein retains its nuclease activity against naked or DNA, yet loses barrier activity. In-frame fusion to D1 of this C-terminal domain creates a chimeric enzyme (D1-D3CT) with significantly increased barrier activity. Substitution of asparagines residues involved in N-linked glycosylation of D1 are known to diminish the targeting of this enzyme to lysosomes [Nishikawa et al. J Biol Chem 1999]. We find that for both D1 and D1-D3CT, N > S substitutions at these residues progressively increase the capacity of the nucleases to block transfection. As expected, the N > S substitutions in murine D1 and chimeric D1-D3CT translated smaller, sharper bands of nuclease activity in SDS-PAGE zymograms. Barrier to transfection activity was quantified by measuring the efficiency of transfection using FuGene-6 (Roche) reagent. of GFP-expression vectors Efficiency was measured by flow cytometry. No GFP expression is detectable by western in D3-exposed cells, and free DNA nuclease activity in the media by RDA was about half that found for D1. As measured by radial diffusion assays, levels of free-DNA nuclease activity in conditioned media were not significantly different between any of the D1 and D1-D3CT enzymes. In summary, D1-D3CT sans N-linked glycosylation erects a more potent a barrier to transfection when compared to both wild-type D1 or D1-D3CT. Results strongly suggest the majority of liposome-bound DNA achieves expression mainly by entering via a non-lysosomal pathway guarded by D3, and generally distinct from that targeted by a lysosomal enzyme like D1. Studies of this nuclease barrier will help explain the mechanism by which liposomal gene transfer occurs, and help devise methods to improve efficiency.

Characterization of the human DNAS1L2 gene and the molecular mechanism for its transcriptional activation induced by inflammatory cytokines

DNAS1L2, a member of the DNase I-like endonuclease family, is the only divalent cation-dependent acid DNase so far identified in mammals. The presence of a proline-rich domain (PRD) is its unique feature among family members. We found that a novel transcript encoding a short product, DNAS1L2-S, is expressed in peripheral blood leukocytes. Although DNAS1L2-S lacks the PRD, its enzymatic properties are apparently the same as those of the previously identified long form, DNAS1L2-L. Sequence analysis reveals that DNAS1L2 consists of seven exons. The exon/intron boundaries agree with the GT/AG rule with one exception: GC replaces GT at the 5′ splice site in the sixth intron. TNF-α and IL-1β are found to be potent inducers of DNAS1L2 expression in keratinocytes. They induce DNAS1L2 activation via the NF-κB pathway, and an NF-κB binding site located within the 5′ flanking region is identified as the cis-responsive element.

High-density labeling of DNA for single molecule sequencing.

Two unusual enzymatic activities are required for the realization of a single molecule sequencing: a polymerase for copying a deoxyribonuclease (DNA) target into complementary flurophore-labeled DNA, and an exonuclease for the successive hydrolysis of the completely dye-labeled DNA. Recently, we found that the wild-type Klenow fragment of Escherichia coli DNA polymerase I is well-suited for the synthesis of DNA in a reaction set-up that contains exclusively specific rhodamine-labeled analogs of the natural pyrimidine nucleotides (dCTP and dTTP). This protocol describes the procedure used for the preparation of DNA that is labeled at all pyrimidine bases of one strand, as well as an example of enzymatic downstream processing of the DNA product.

Sodium nitroprusside and peroxynitrite effect on hepatic DNases: an in vitro and in vivo study

BackgroundIt has been documented that nitric oxide (NO) donor sodium nitroprusside (SNP) and authentic peroxynitrite are capable of promoting apoptosis in a number of different cell types. Various endonucleases have been proposed as candidates responsible for the internucleosomal cleavage of the genomic DNA observed during apoptosis, but the main effect is attributed to the alkaline-DNases (Mg2+- and caspase-dependent) and acid-DNase. The aim of this study was to examine an in vivo and in vitro possibility for alkaline- and acid-DNases to be activated by SNP and peroxynitrite.ResultsThe effect on liver tissue alkaline and acid DNase activity together with the markers of tissue and plasma oxidative and nitrosative stress (lipid peroxidation, SH group content, carbonyl groups and nitrotyrosine formation) was investigated in plasma and liver tissue. The activity of liver alkaline DNase increased and that of acid DNase decreased after in vivo treatment with either SNP or peroxynitrite. A difference observed between the in vivo and in vitro effect of oxide donor (i.e., SNP) or peroxynitrite upon alkaline DNase activity existed, and it may be due to the existence of the "inducible" endonuclease. After a spectrophotometric scan analysis of purified DNA, it was documented that both SNP and peroxynitrite induce various DNA modifications (nitroguanine formation being the most important one) whereas DNA fragmentation was not significantly increased.ConclusionAlkaline DNase activation seems to be associated with the programmed destruction of the genome, leading to the fragmentation of damaged DNA sites. Thus, the elimination of damaged cells appears to be a likely factor in prevention against mutation and carcinogenesis.

Non-classic characteristics define prominent DNase activities from the intestine and other tissues of Haemonchus contortus

The anthelmintic fenbendazole (FBZ) induces nuclear DNA fragmentation (DF) in intestinal cells of Haemonchus contortus. The DNA fragments had 3 ′-OH, which suggests involvement of a neutral DNase. To identify candidate DNase(s) involved, DNase activity in H. contortus intestine and other worm fractions was characterized relative to classic DNases I (neutral) and II (acidic). Seven distinct DNase activities were identified and had M rs of 34, 36, 37 or 38.5 kDa on zymographic analysis. The different activities were distinguished according to pH requirement, sensitivity to 10 mM EDTA and worm compartment. Activities of intestinal DNases at 34, 36 and 38.5 kDa were sensitive to EDTA at pH 5.0 and 7.0. Sensitivity to EDTA at pH 5.0 was unexpected compared to classic acidic DNase II activity, suggesting unusual properties of these DNases. In whole worms, however, the activities at 36 and 38.5 kDa were relatively insensitive to EDTA, indicating predominance of DNases that are distinct from the intestine. The activity at 37 kDa in excretory/secretory products had an acidic pH requirement and was insensitive to EDTA, resembling classic acidic DNase activity. Under conditions of pH 5.0 and 7.0, intestinal DNases produced 3 ′-ends that could be labeled by terminal deoxynucleotidyl transferase, indicating presence of 3 ′-OH. The labeling of 3 ′-ends at pH 5.0, again, was unexpected for acidic DNase activity. These results and several other activities suggest that multiple H. contortus DNases have characteristics distinct from the classic mammalian DNases I and II. Treatment of H. contortus with FBZ did not induce any detectable DNase activities distinct from normal intestine, although relative activities of intestinal DNases appear to have been altered by this treatment. Abbreviation: DNase, deoxyribonuclease; DF, DNA fragmentation; BZ, benzimidazole anthelmintics; TdT, terminal deoxynucleotidyl transferase; ATA, aurintricarboxylic acid

Nuclear cataract caused by a lack of DNA degradation in the mouse eye lens.

The eye lens is composed of fibre cells, which develop from the epithelial cells on the anterior surface of the lens. Differentiation into a lens fibre cell is accompanied by changes in cell shape, the expression of crystallins and the degradation of cellular organelles. The loss of organelles is believed to ensure the transparency of the lens, but the molecular mechanism behind this process is not known. Here we show that DLAD ('DNase II-like acid DNase', also called DNase IIbeta) is expressed in human and murine lens cells, and that mice deficient in the DLAD gene are incapable of degrading DNA during lens cell differentiation--the undigested DNA accumulates in the fibre cells. The DLAD-/- mice develop cataracts of the nucleus lentis, and their response to light on electroretinograms is severely reduced. These results indicate that DLAD is responsible for the degradation of nuclear DNA during lens cell differentiation, and that if DNA is left undigested in the lens, it causes cataracts of the nucleus lentis, blocking the light path.

Isolation and Identification of Staphyococci from Raw Milk of Cows Infected with Mastitis

Twenty-six isolates of staphylococci were recovered from of 100 raw milk samples collected from one hundred cows infected with mastitis. For the isolation of bacteria, Mannitol salt agar was used as a selective medium. Staphylococcal species were identified by API-STAPH system. The identified species were: - Staphy aureus (13), Staph. epidermidis(9), Staph. haemolyticus(2), Staph. lugdunensis()), and Staph. hominis). Staphylococcus aureus was the most prevalent pathogen among other Staphyloccocal species isolated in this study. Staph. aureus isolates revealed positive results for the defection of capsule structure and for production of some essential enzymes such as coagulase, phosphatase, DNA ase and haemolysin which are highly associated with the virulance of bacteria. Staph. aureus isolates were tested for their sensitivity to the antibiotics and the results were: 13(100%) isolates were sensitive to Gentamycin and Tetracyclene, 11 (84.6%) isolates were sensitive to Oxacillin, Penicillin, Ampicillin, Erythromycin and Cephalexin, where as only 2 (15.4%) isolates were Methicillin Resistant Staph. aureus (MRSA) which showed multi -resistant 
 towards many antibiotics used in this work.