Jpn J Infect Dis, 2004 Oct, 57(5), S12 - 3 Genomic variations in myeloperoxidase gene in the Japanese population; Kameoka Y et al.; Myeloperoxidase (MPO; EC 1.11.1.7) is a lysosomal hemeprotein that plays an important role in the host defense mechanism against microbial diseases . This neutrophil disorder, characterized by the lack of MPO, may result in a weakened defense activity . Complete MPO deficiency has been postulated to be to originate from genomic mutation . Recently, two Japanese patients were reported with MPO deficiency . Both had base substitutions in the exon 9 region of the MPO gene; a region in close proximity functionally important residue, His502 . Genomic DNA from 387 Japanese individuals was examined to determine the prevalence of these recently discovered base substitutions . None of these DNA samples possessed the mutations found in the MPO deficient cases, though two synonymous and one non-synonymous mutation were found . The frequency of mutation in the exon 9 coding region was estimated to be one heterozygote in 129, thus the homozygote of such mutations would be revealed one in 16,000 in the Japanese population. J Virol, 2004 Nov, 78(22), 12344 - 54 CpG oligodeoxynucleotides block human immunodeficiency virus type 1 replication in human lymphoid tissue infected ex vivo; Schlaepfer E et al.; Oligodeoxynucleotides (ODNs) with immunomodulatory motifs control a number of microbial infections in animal models, presumably by acting through toll-like receptor 9 (TLR9) to induce a number of cytokines (e.g., alpha interferon and tumor necrosis factor alpha) . The immunomodulatory motif consists of unmethylated sequences of cytosine and guanosine (CpG motif) . ODNs without CpG motifs do not trigger TLR9 . We hypothesized that triggering of TLR9 generates a cellular environment unfavorable for human immunodeficiency virus (HIV) replication . We tested this hypothesis in human lymphocyte cultures and found that phosphorothioate-modified ODN CpG2006 (type B ODNs) inhibited HIV replication nearly completely and prevented the loss of CD4(+) T cells . ODNs CpG2216 and CpG10 (type A ODNs) were less effective . CpG2006 blocked HIV replication in purified CD4(+) T cells and T-cell lines; CpG10 was ineffective in this setting, indicating that type A ODNs may inhibit HIV replication in CD4(+) T-cell lines indirectly through a separate cell subset . However, control ODNs without CpG motifs also showed anti-HIV effects, indicating that these effects are nonspecific and not due to TLR9 triggering . The mechanism of action is not clear . CpG2006 and its control ODN blocked syncytium formation in a cell fusion-based assay, but CpG10, CpG2216, and their control ODNs did not . The latter types interfered with the HIV replication cycle during disassembly or reverse transcription . In contrast, CpG2006 and CpG2216 specifically induced cytokines critical to initiation of the innate immune response . In summary, the nonspecific anti-HIV activity of CpG ODNs, their ability to stimulate HIV replication in latently infected cells, potentially resulting in their elimination, and their documented ability to link the innate and adaptive immune responses make them attractive candidates for further study as anti-HIV drugs. J Biol Chem, 2005 Jan 7, 280(1), 571 - 7 Epub 2004 Oct 26. A Unique Hydrophobic Cluster Near the Active Site Contributes to Differences in Borrelidin Inhibition among Threonyl-tRNA Synthetases; Ruan B et al.; Borrelidin, a compound with anti-microbial and anti-angiogenic properties, is a known inhibitor of bacterial and eukaryal threonyl-tRNA synthetase (ThrRS) . The inhibition mechanism of borrelidin is not well understood . Archaea contain archaeal and bacterial genre ThrRS enzymes that can be distinguished by their sequence . We explored species-specific borrelidin inhibition of ThrRSs . The activity of ThrRS from Sulfolobus solfataricus and Halobacterium sp . NRC-1 was inhibited by borrelidin, whereas ThrRS enzymes from Methanocaldococcus jannaschii and Archaeoglobus fulgidus were not . In Escherichia coli ThrRS, borrelidin binding induced a conformational change, and threonine was not activated as shown by ATP-PP(i) exchange and a transient kinetic assay measuring intrinsic tryptophan fluorescence changes . These assays further showed that borrelidin is a noncompetitive tight binding inhibitor of E . coli ThrRS with respect to threonine and ATP . Genetic selection of borrelidin-resistant mutants showed that borrelidin binds to a hydrophobic region (Thr-307, His-309, Cys-334, Pro-335, Leu-489, Leu-493) proximal to the zinc ion at the active site of the E . coli ThrRS . Mutating residue Leu-489 --> Trp reduced the space of the hydrophobic cluster and resulted in a 1500-fold increase of the K(i) value from 4 nm to 6 mum . An alignment of ThrRS sequences showed that this cluster is conserved in most organisms except for some Archaea (e.g . M . jannaschii, A . fulgidus) and some pathogens (e.g . Helicobacter pylori) . This study illustrates how one class of natural product inhibitors affects aminoacyl-tRNA synthetase function, providing potentially useful information for structure-based inhibitor design. Genes Cells, 2004 Nov, 9(11), 1055 - 67 ASC is essential for LPS-induced activation of procaspase-1 independently of TLR-associated signal adaptor molecules; Yamamoto M et al.; Toll-like receptors (TLRs) initiate a signalling cascade via association with an adaptor molecule, myeloid differentiation factor 88 (MyD88) and/or TIR domain-containing adaptor inducing-IFN-beta (Trif), to induce various pro-inflammatory cytokines for microbial eradication . After stimulation of TLR4 with lipopolysaccharide (LPS), both IL-1beta and IL-18 are processed, depending on the activation of caspase-1, although its mechanism remains unclear . ASC is an adapter protein possibly involved in the activation of procaspase-1 . To unravel the requirement of ASC, we generated Asc(-/-) mice . Upon stimulation with LPS, Asc(-/-) macrophages failed in the processing of procaspase-1 and maturation of pro-IL-1beta and pro-IL-18, but normally produced other pro-inflammatory cytokines including TNF-alpha and IL-6 . MyD88(-/-) and Trif(-/-) macrophages showed normal activation of caspase-1, demonstrating a dispensable role for MyD88 and Trif . After, LPS-challenged Asc(-/-) mice lacked serum elevation of IL-1beta and IL-18 . Moreover, the Asc(-/-) mice exhibited neither acute liver injury nor lethal shock . These results demonstrate critical roles for ASC in the release of IL-1beta/IL-18 via activation of caspase-1 and provide new insights into the inflammatory responses for host defence and diseases. Environ Sci Technol, 2004 Oct 1, 38(19), 5134 - 40 Azo dye method for mapping relative sediment enzyme activity in situ at precise spatial locations; Rogers NJ et al.; Existing methodology for measuring microbial enzyme activity in aquatic sediments involves horizontal sectioning of sediment cores into centimeter slices, followed by determination of the enzyme activity of each homogenized sediment slice . At best, this approach provides only one-dimensional information on the distribution of microbial activity . This paper describes the development of a novel technique to map sediment enzyme activity in situ at millimeter spatial resolution . Naphthol AS enzyme substrates were loaded onto filter membranes by evaporation from an organic solvent . The membranes were attached to plastic cards to form rigid probes, which were deployed vertically in sediments for a fixed time period . The exposed membranes were developed in a diazonium salt solution, resulting in the formation of a colored precipitate where substrate hydrolysis had occurred . The chromogenic reaction was calibrated and quantified by immersing substrate-loaded membranes in a series of solutions of known enzyme activity . A flatbed scanner and image analysis software were used to produce digitized images and to generate two-dimensional maps of enzyme activity . The technique was used to map the spatial features of esterase activity in aquatic sediment samples from wetland areas and enabled the precise locations of microbial activity "hotspots" to be identified. Ying Yong Sheng Tai Xue Bao, 2004 Jul, 15(7), 1227 - 30 {Effect of fertilization on cucumber growth and soil biological characteristics in sunlight greenhouse}; Xu F et al.; This paper studied the effect of fertilization on cucumber growth and yield, soil microbial biomass and soil enzyme activities in sunlight greenhouse in Loess Plateau . The results indicated that the growth and yield of cucumber were increased with application of manure and methane . Foliage application reduced the application rate of NP and manure . Fertilization had an obvious effect on the biological characteristics of soil in sunlight greenhouse . The number of bacteria was increased by manure and foliage fertilization, and that of fungi was increased by NP and methane fertilization but decreased by manure fertilization . Fertilization with manure, NP and methane also remarkably increased the number of actinomyces and the activities of urease, phosphatase and sucrase in soil . The activities of soil urease and phosphatase were increased by fertilization of manure, but little effect was found with fertilization of NP and methane. Apoptosis, 2004 Nov, 9(6), 729 - 37 Proteinase-activated receptor 1 (PAR-1) and cell apoptosis; Flynn AN et al.; This review summarizes the main aspects and newest findings of how proteinase-activated receptor 1 (PAR-1) may modulate programmed cell death . Activation of PAR-1 has been found to induce or inhibit apoptosis in a variety of cells, depending on the dosage of its physiological agonist thrombin, or that of synthetic receptor activators . To date, cellular targets for PAR-1-mediated effects on apoptosis include neuronal, endothelial, and epithelial cells, fibroblasts, and tumor cells . The signaling pathways involved in the induction or prevention of apoptosis by PAR-1 activation are diverse, and include JAK/STAT, RhoA, myosin light chain kinase, ERK1/2, and various Bcl-2 family members . In view of the well-established involvement of microbial proteinases in host tissue malfunction, the article also elaborates on the possible significance of PAR-1 activation for the pathogenesis of infectious disease. Proc Natl Acad Sci U S A, 2004 Nov 2, 101(44), 15718 - 23 Epub 2004 Oct 25. The gut microbiota as an environmental factor that regulates fat storage; Backhed F et al.; New therapeutic targets for noncognitive reductions in energy intake, absorption, or storage are crucial given the worldwide epidemic of obesity . The gut microbial community (microbiota) is essential for processing dietary polysaccharides . We found that conventionalization of adult germ-free (GF) C57BL/6 mice with a normal microbiota harvested from the distal intestine (cecum) of conventionally raised animals produces a 60% increase in body fat content and insulin resistance within 14 days despite reduced food intake . Studies of GF and conventionalized mice revealed that the microbiota promotes absorption of monosaccharides from the gut lumen, with resulting induction of de novo hepatic lipogenesis . Fasting-induced adipocyte factor (Fiaf), a member of the angiopoietin-like family of proteins, is selectively suppressed in the intestinal epithelium of normal mice by conventionalization . Analysis of GF and conventionalized, normal and Fiaf knockout mice established that Fiaf is a circulating lipoprotein lipase inhibitor and that its suppression is essential for the microbiota-induced deposition of triglycerides in adipocytes . Studies of Rag1-/- animals indicate that these host responses do not require mature lymphocytes . Our findings suggest that the gut microbiota is an important environmental factor that affects energy harvest from the diet and energy storage in the host . Data deposition: The sequences reported in this paper have been deposited in the GenBank database (accession nos . AY 667702--AY 668946). J Exp Med, 2004 Nov 1, 200(9), 1213 - 9 Epub 2004 Oct 25. Siderophore biosynthesis but not reductive iron assimilation is essential for Aspergillus fumigatus virulence; Schrettl M et al.; The ability to acquire iron in vivo is essential for most microbial pathogens . Here we show that Aspergillus fumigatus does not have specific mechanisms for the utilization of host iron sources . However, it does have functional siderophore-assisted iron mobilization and reductive iron assimilation systems, both of which are induced upon iron deprivation . Abrogation of reductive iron assimilation, by inactivation of the high affinity iron permease (FtrA), has no effect on virulence in a murine model of invasive aspergillosis . In striking contrast, A . fumigatus L-ornithine-N5-monooxygenase (SidA), which catalyses the first committed step of hydroxamate-type siderophore biosynthesis, is absolutely essential for virulence . Thus, A . fumigatus SidA is an essential virulence attribute . Combined with the absence of a sidA ortholog-and the fungal siderophore system in general-in mammals, these data demonstrate that the siderophore biosynthetic pathway represents a promising new target for the development of antifungal therapies. Transfusion, 2004 Nov, 44(11), 1588 - 92 Altered glycosylation leads to Tr polyagglutination; Halverson GR et al.; BACKGROUND: Polyagglutination refers to red blood cells (RBCs) that are agglutinated by a high proportion of ABO-matched adult sera but not by cord sera . Polyagglutinable RBCs have been associated with microbial infection, myeloproliferative disorders, and myelodysplasia . Lectins aid in the identification of polyagglutination . CASE STUDY: A Hispanic male infant with mild hemolytic anemia, a "Bernard-Soulier-like" syndrome, intermittent neutropenia, mitral valve regurgitation, ligament hyperlaxity, and mild mental retardation was studied . The patient's Group O RBCs were polyagglutinable; they were agglutinated by normal human sera, several lectins {including Arachis hypogea, Salvia sclarea, Salvia horminum, Glycine max, Ulex europaeus, Griffonia simplicifolia I, and Gr . simplicifolia II}, and some monoclonal antibodies . His RBCs were not agglutinated by cord sera, Dolichos biflorus, or Phaseolus lunatus . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis on the RBC membranes followed by staining with periodic acid-Schiff stain showed markedly reduced staining of glycophorins A and B . Staining with Coomassie brilliant blue revealed that Band 3 has a faster mobility than normal . CONCLUSIONS: Collectively, the results suggest that the patient's RBCs have a reduction in N-acetylneuraminic acid on both N- and O-glycans, exposing, respectively, beta1,4-galactosidase and beta1,3-galactosidase . The patient likely has an altered glycosyltransferase that results in defective glycosylation in RBCs and other cell lineages . This type of polyagglutination was named Tr. Refuat Hapeh Vehashinayim, 2004 Apr, 21(2), 7 - 16, 95 Apical problems--coronal solutions: how changes in access cavity design can prevent and overcome problems with curved canals; Druttman AC; There is no doubt that the preparation of curved canals presents one of the greatest challenges in endodontic and is fraught with potential difficulties . Canal curvature can only be seen on a radiograph in the mesio-distal plane and yet it is well known that curvature in the bucco-lingual plane is also evident in many teeth . A primary cause of failure of endodontic treatment is the persistence of microbial infection . In curved canals this is often due to procedural errors such as ledges, fractured instruments and canal blockage . Over the years a variety of techniques have been proposed for preparing these canals . Whether it is the straightforward endodontic treatment of a straight, single rooted tooth, or retreatment of a curved molar with a number of procedural errors, the access cavity has a crucial role in the achievement of successful endodontic treatment . The size and shape of the access cavity will be dictated by the degree of curvature of the canals and the objectives that have to be achieved. Ophthalmologe . 2004 Oct 20; {Epub ahead of print} {Microbial keratitis following laser in situ keratomileusisPrevention, differential diagnosis, and therapy.}; Tehrani M et al.; Laser in situ keratomileusis (LASIK) is an effective option and currently one of the most commonly applied surgical techniques in the correction of refractive errors such as myopia, hyperopia, and astigmatism . In contrast to photorefractive keratectomy, it maintains the integrity of Bowman's membrane and the epithelium leading to faster visual rehabilitation as well as less pain and discomfort . Nevertheless, following LASIK the stroma is exposed to infectious organisms . Sight-threatening complications after LASIK are reported to be as rare as 1 in 1000 procedures . However, any infectious keratitis remains potentially devastating . Reports about infectious keratitis following LASIK have increasingly surfaced in recent years . We present a review of the literature on microbial keratitis and present our own cases and recommendations for possible prophylaxis and therapy. AJNR Am J Neuroradiol, 2004 Oct, 25(9), 1544 - 8 An unusual cause of visual loss: involvement of bilateral lateral geniculate bodies; Lefebvre PR et al.; We report the clinical and radiologic features of a 31-year-old woman who suffered incongruous binasal and bitemporal visual field defects and severe sudden visual loss due to hypoperfusion of bilateral lateral geniculate bodies following anaphylactic shock induced by 500 mg amoxicillin per os . Complete neuroophthalmologic examinations were performed regularly for visual acuity, color vision, pupillary reflexes, and visual fields . Additional testing was performed by means of MR imaging of the brain and CSF analysis . Follow-up was performed for 12 months . Vision loss was acute and severe, its onset bilateral and simultaneous . The patient recovered visual acuity of 1.0 within 7 weeks . Color vision was abnormal in both eyes but gradually improved to normal . Visual fields were characterized by incongruous binasal and bitemporal defects, but they reduced progressively . Cerebral MR imaging confirmed the presence of symmetrical lesions confined exclusively within both lateral geniculate bodies . These lesions were best seen on T1-weighted and fluid-attenuated inversion recovery images as high-signal-intensity areas suggestive of hemorrhagic ischemia . CSF analysis was normal and aseptic . Blood tests and cultures excluded any microbial infection . We conclude that shock may induce a bilateral isolated ischemia of the lateral geniculate bodies, resulting in incongruous binasal and bitemporal visual field defects and severe visual loss . MR imaging is the optimal imaging technique to confirm the diagnosis and for follow-up . Copyright American Society of Neuroradiology Peptides, 2004 Nov, 25(11), 1873 - 81 Efficacy of human beta-casein fragment (54-59) and its synthetic analogue compound 89/215 against Leishmania donovani in hamsters; Sharma P et al.; The characteristic feature of visceral leishmaniasis (VL) is the profound impairment of immune system of the infected host, which contributes significantly to the partial success of antileishmanial chemotherapy . Since in VL, cure is the combinatorial effect of drug and immune status of the host, the rationale approach towards antileishmanial chemotherapy would be to potentiate the immune functioning of the host to extract desired results . Towards this direction several rationally designed analogues of human beta-casein fragment (54-59) were evaluated for their ability to stimulate the non-specific resistance in hamsters against Leishmania donovani infection . By virtue of being derived from the food protein casein derivatives may be devoid of unwanted side effects associated with the substances of microbial origin, e.g . muramyl dipeptide (MDP) . Out of this one peptide Val-Glu-Gly-Ile-Pro-Tyr (compound 89/215) had been reported to have such activity . In this communication, the prophylactic and therapeutic efficacy of the peptide along with its natural sequence has been evaluated in detail against experimental VL in hamsters . Their use as an adjunct to chemotherapy was also explored . Human beta-casein fragment, compound 89/215 and MDP were tested in vivo at various dose levels wherein compound 89/215 showed superiority over MDP at 3 mg/kg x 2 given intraperitoneally (i.p.) . Compound 89/215 sensitized peritoneal macrophages acquired considerable resistance and only 24% of the cells were found infected in comparison to control peritoneal macrophages where 76.4% of the cells were found infected . Similarly, the efficacy of sodium antimony gluconate (SAG) in hamsters pretreated with compound 89/215 enhanced significantly (P < 0.001) . This peptide also exhibited considerably good therapeutic efficacy when evaluated either alone or in combination with SAG in established infection of L . donovani. Toxicon, 2004 Nov, 44(6), 581 - 8 The beta-defensin-fold family of polypeptides; Torres AM et al.; Polypeptides adopting a fold very similar to that of beta-defensins are found in diverse organisms, including sea anemones, snakes, platypus and humans . These molecules of approximately 35-50 amino acid residues possess disparate activities, such as anti-microbial, myonecrotic, analgesic, and ion-channel inhibiting . The family of beta-defensin-fold structures generally consists of a short helix or turn followed by a small twisted anti-parallel beta-sheet . The six cysteine residues which are paired in a 1-5, 2-4, 3-6 fashion are crucial for determining and maintaining the compact core configuration of the structures . The primary structural similarity between members of the family suggests that the global fold is robust and that the nature of the side-chains determine the functional specificity . The distinct compact fold shared by these polypeptides may be useful in the design of molecules with desired pharmacological activity. Expert Opin Biol Ther, 2004 Nov, 4(11), 1719 - 27 Biological response modifiers for the treatment of Crohn's disease; Siveke JT et al.; Although standard medical therapy in Crohn's disease is efficient in most patients, a substantial proportion of patients suffering from chronic active disease do not adequately respond to standard therapy . In these patients, alternative regimens have to be considered . Due to the major advances in understanding the pathogenesis of this complex disease involving genetic, environmental, microbial and immunological factors, various new biological therapies targeting key mechanisms have emerged . In this review, a critical appraisal of modern therapeutical concepts will be presented, focusing on antibody and small inhibitory molecule therapies, including inhibition of TNF-alpha and other pro-inflammatory cytokines, adhesion molecules and T cell activation, as well as hormonal therapies. J Infect Dis, 2004 Nov 15, 190(10), 1767 - 72 Epub 2004 Oct 07. Link between impaired maturation of phagosomes and defective Coxiella burnetii killing in patients with chronic Q fever; Ghigo E et al.; Q fever is caused by Coxiella burnetii, a bacterium that survives in monocytes/macrophages by resisting their natural microbicidal activity . Because the link between bacterial killing and phagosome maturation has yet to be demonstrated, we evaluated responses in monocytes from both immunologically naive control subjects and patients with various manifestations of Q fever . Monocytes from patients with chronic Q fever in evolution, who do not control the infection, exhibited defective phagosome maturation and impaired C . burnetii killing . Both responses were stimulated in patients recovering from Q fever . Phagosome maturation and C . burnetii killing were significantly correlated . Defective phagosome maturation and impaired C . burnetii killing were induced by adding interleukin (IL)-10 to monocytes from convalescent patients and were restored by IL-10 neutralization in chronic Q fever in evolution . We show that phagosome maturation and microbial killing are linked in Q fever and that IL-10 regulates both features of microbicidal activity. Arch Environ Contam Toxicol, 2004 Nov, 47(4), 561 - 8 Characterization of soils containing adipocere; Fiedler S et al.; The formation of adipocere (commonly known as grave wax), a spontaneous inhibition of postmortem changes, has been extensively analyzed in forensic science . However, soils in which adipocere formation occurs have never been described in detail . Therefore, this study is intended as a first step in the characterization of soils containing adipocere . Two grave soils (Gleyic Anthrosols) that prevent the timely reuse of graves due to the occurrence of adipocere and a control soil (Gleyic Luvisol) were selected from a cemetery in the Central Black Forest (Southwest Germany) . Descriptions of soil morphology and a wide assay of physical, chemical, and microbiologic soil characteristics were accomplished . In contrast to the control soil, the grave soils were characterized by lower bulk density and pH . The degradation of the soil structure caused by digging led to a higher water table and the expansion of the reducing conditions in the graves where the prevalent absence of oxygen in range of the coffins inhibited decomposition processes . Although the formation of adipocere led to the conservation of the buried corpses, phosphorus, dissolved organic carbon, and cadavarine leaching from the graves was observed . Microbial biomass and microbial activity were higher in the control soil and hence reflected the inert character of adipocere . The study results clearly show the need for additional approaches in forensic, pedologic, and microbiologic research. Arch Environ Contam Toxicol, 2004 Nov, 47(4), 440 - 7 Use of zeolite for removing ammonia and ammonia-caused toxicity in marine toxicity identification evaluations; Burgess RM et al.; Ammonia occurs in marine waters including effluents, receiving waters, and sediment interstitial waters . At sufficiently high concentrations, ammonia can be toxic to aquatic species . Toxicity identification evaluation (TIE) methods provide researchers with tools for identifying aquatic toxicants . For identifying ammonia toxicity, there are several possible methods including pH alteration and volatilization, Ulva lactuca addition, microbial degradation, and zeolite addition . Zeolite addition has been used successfully in freshwater systems to decrease ammonia concentrations and toxicity for several decades . However, zeolite in marine systems has been used less because ions in the seawater interfere with zeolite's ability to adsorb ammonia . The objective of this study was to develop a zeolite method for removing ammonia from marine waters . To accomplish this objective, we performed a series of zeolite slurry and column chromatography studies to determine uptake rate and capacity and to evaluate the effects of salinity and pH on ammonia removal . We also assessed the interaction of zeolite with several toxic metals . Success of the methods was also evaluated by measuring toxicity to two marine species: the mysid Americamysis bahia and the amphipod Ampelisca abdita . Column chromatography proved to be effective at removing a wide range of ammonia concentrations under several experimental conditions . Conversely, the slurry method was inconsistent and variable in its overall performance in removing ammonia and cannot be recommended . The metals copper, lead, and zinc were removed by zeolite in both the slurry and column treatments . The zeolite column was successful in removing ammonia toxicity for both the mysid and the amphipod, whereas the slurry was less effective . This study demonstrated that zeolite column chromatography is a useful tool for conducting marine water TIEs to decrease ammonia concentrations and characterize toxicity. Eye Contact Lens, 2004 Oct, 30(4), 231 - 3; discussion 242-3 Adverse corneal events associated with corneal reshaping: a case series; Lang J et al.; PURPOSE: This case series presents the first documented cases of infectious ulcers associated with overnight orthokeratology in North America and other less serious complications associated with overnight corneal reshaping . CASE REPORTS: Five cases of adverse corneal events associated with corneal refractive therapy are described: two cases of microbial keratitis, one case of infiltrates, one case of toxic keratitis, and one corneal abrasion . CONCLUSIONS: Corneal compromise and poor compliance can cause adverse events with corneal reshaping . The need for ongoing patient education is important not only for pediatric contact lens patients, but also for adults. J Leukoc Biol, 2005 Jan, 77(1), 67 - 70 Epub 2004 Oct 21. Human V{gamma}9/V{delta}2 effector memory T cells express the killer cell lectin-like receptor G1 (KLRG1); Eberl M et al.; The killer cell lectin-like receptor G1 (KLRG1) is expressed in natural killer (NK) cells and effector memory alphabeta T cells . gammadelta T cells represent an unconventional lymphocyte population that shares characteristics of NK cells and T cells and links innate and adaptive immunity . Vgamma9/Vdelta2 T cells comprise the majority of peripheral human gammadelta T cells and respond to the microbial metabolite (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMB-PP) . Here, we demonstrate that KLRG1 is expressed in a significant proportion of Vgamma9/Vdelta2 T cells in cord blood and in the majority of peripheral Vgamma9/Vdelta2 T cells from adult donors . KLRG1(+) Vgamma9/Vdelta2 T cells displayed an effector memory phenotype, as KLRG1 was expressed mainly in Vgamma9/Vdelta2 T cells lacking CD27, CD45RA, CD62L, and CC chemokine receptor 7 (CCR7) . Unlike alphabeta T cells, where possession of KLRG1 identified effector memory cells with impaired proliferative capacity, KLRG1(+) Vgamma9/Vdelta2 T cells were able to proliferate vigorously upon stimulation with HMB-PP in the presence of interleukin-2 . Moreover, KLRG1 ligation on Vgamma9/Vdelta2 T cells by antibodies did not inhibit HMB-PP-induced proliferation and cytokine production nor cytolysis of Daudi cells. Mol Nutr Food Res, 2004 Sep, 48(4), 326 - 32 The pig caecum model: a suitable tool to study the intestinal metabolism of flavonoids; Labib S et al.; Pig caecum was used under anaerobic conditions to metabolize flavonoids from several classes, i.e., chrysin 1, naringenin 2, quercetin 3, and hesperetin 4 . Whereas chrysin 1 was not converted by the pig intestinal flora under the experimental conditions used, naringenin 2 was transformed to 3-(4-hydroxyphenyl)-propionic acid and 3-phenylpropionic acid . Quercetin 3 was metabolized to phloroglucinol, 3,4-dihydroxyphenylacetic acid, and 3,4-dihydroxytoluene . Hesperetin 4 was degraded via eriodictyol to 3-(3-hydroxyphenyl)-propionic acid and phloroglucinol . Structural elucidation of the formed metabolites was performed by high-performance liquid chromatography--diode array detection (HPLC-DAD) as well as HPLC-electrospray ionization--mass spectrometry (ESI-MS (MS)) and high resolution gas chromatography-mass spectrometry (HRGC-MS) analyses . The time course of microbial conversion of 2-4 was determined by HPLC-DAD analysis, revealing slow degradation of 2 and rapid transformation of 3 and 4 . The results lead to the conclusion that the pig caecum model is a suitable ex vivo model for studying the intestinal degradation of flavonoids. J Immunol, 2004 Nov 1, 173(9), 5688 - 96 Expression of functional TLR4 confers proinflammatory responsiveness to Trypanosoma cruzi glycoinositolphospholipids and higher resistance to infection with T . cruzi; Oliveira AC et al.; TLRs function as pattern recognition receptors in mammals and play an essential role in the recognition of microbial components . We found that the injection of glycoinositolphospholipids (GIPLs) from Trypanosoma cruzi into the peritoneal cavity of mice induced neutrophil recruitment in a TLR4-dependent manner: the injection of GIPL in the TLR4-deficient strain of mice (C57BL/10ScCr) caused no inflammatory response . In contrast, in TLR2 knockout mice, neutrophil chemoattraction did not differ significantly from that seen in wild-type controls . GIPL-induced neutrophil attraction and MIP-2 production were also severely affected in TLR4-mutant C3H/HeJ mice . The role of TLR4 was confirmed in vitro by testing genetically engineered mutants derived from TLR2-deficient Chinese hamster ovary (CHO)-K1 fibroblasts that were transfected with CD14 (CHO/CD14) . Wild-type CHO/CD14 cells express the hamster TLR4 molecule and the mutant line, in addition, expresses a nonfunctional form of MD-2 . In comparison to wild-type cells, mutant CHO/CD14 cells failed to respond to GIPLs, indicating a necessity for a functional TLR4/MD-2 complex in GIPL-induced NF-kappaB activation . Finally, we found that TLR4-mutant mice were hypersusceptible to T . cruzi infection, as evidenced by a higher parasitemia and earlier mortality . These results demonstrate that natural resistance to T . cruzi is TLR4 dependent, most likely due to TLR4 recognition of their GIPLs. Bioresour Technol, 2005 Mar, 96(4), 479 - 84 Characterization of natural organic matter (NOM) derived from sewage sludge compost . Part 2: multivariate techniques in the study of compost maturation; Zbytniewski R et al.; This study presents the results of chemometric data analysis which describes the maturation of sewage sludge compost . The compost was characterized at different stages of maturation by various chemical and spectroscopic parameters including carbon and nitrogen content, humic substances content, UV-Vis and (13)C NMR . The data set of compost characteristics was analyzed using multivariate methods: cluster analysis (CA) and principal component analysis (PCA) . The results enabled the determination of three groups of compost samples at different stages of maturation that correspond to three stages of composting: (i) domination of rapid decomposition of non-humic, easily biodegradable organic matter, (ii) domination of organic matter humification and formation of polycondensed, humic-like substances (the next 2 weeks), (iii) stabilization of transformed organic material and weak microbial activity . The multivariate techniques also enabled the identification of main parameters that change during different stages of composting the most. Bioresour Technol, 2005 Mar, 96(4), 471 - 8 Characterization of natural organic matter (NOM) derived from sewage sludge compost . Part 1: chemical and spectroscopic properties; Zbytniewski RA et al.; In this study changes in the properties of natural organic matter (NOM) were studied during composting of sewage sludge in a laboratory experiment using the pile method . Typical physicochemical parameters were measured during 53 days of composting including humic fractions . The effects of humification on the molecular properties of humic acids (HA) were investigated by {Formula: see text} C CP/MAS NMR spectroscopy . On the basis of chemical analyses, 53 days of composting sewage sludge with structural material can be divided into three phases: (i) domination of rapid decomposition of non-humic, easily biodegradable organic matter (two to three weeks), (ii) domination of organic matter humification and formation of polycondensed, humic-like substances (the next two weeks), (iii) stabilization of transformed organic material and weak microbial activity . Spectroscopic characterization ( {Formula: see text} C NMR) of compost humic acids reveals changes in their structures during maturation . The changes are highly correlated with the processes taking place in bulk compost. Water Res, 2004 Nov, 38(19), 4173 - 85 Fate of perchlorate-contaminated water in upflow wetlands; Tan K et al.; The potential of natural wetland systems to treat perchlorate-contaminated water was investigated in vertical upflow wetland columns planted with and without Bulrush (Scirpus sp.) . In the absence of nitrate (NO3- -N <1mg/L), wetland columns were capable of removing ClO4- to levels below the detection limit (<4 microg/L) for a series of influent ClO4- (4, 8, 16, and 32 mg/L) . At an influent ClO4- concentration of 32 mg/L, ClO4- breakthrough was observed with the increase in nitrate concentration . ClO4- and NO3- degradation rate constants (Kpc and KNO3- -N) were also determined using a 1-D transport model with dispersion . Kpc declined with the increase of influent ClO4- and NO3- -N concentration (6.49-0.42 day(-1) for unplanted columns, and 7.80-0.21 day(-1) for planted columns, respectively) . KNO3- -N followed similar trends but was relatively higher than Kpc . Plant uptake was directly linked with ClO4- concentration in the rhizosphere, and the stem bio-concentration factor (BCF) was estimated to be 57 . A mass balance indicated plant uptake accounted for 0-14.3% of initial ClO4- input . Microbial degradation played a more important role than plant uptake and transformation in ClO4- degradation in this wetland system . This study suggests that constructed wetlands may be a promising technology to treat perchlorate-contaminated waters. Water Res, 2004 Nov, 38(19), 4125 - 34 Pilot studies for in-situ aerobic cometabolism of trichloroethylene using toluene-vapor as the primary substrate; Tom Kuo MC et al.; In-situ pilot studies of aerobic cometabolism were conducted to evaluate the injection of toluene-vapor and air into TCE-contaminated aquifer . Delivery of primary substrate (toluene) in a vapor state with air enhanced the growth of indigenous toluene-utilizing bacteria that would degrade TCE by aerobic cometabolism . Meanwhile, delivering toluene in a vapor state effectively reduced potential clogging near the injection points due to excessive microbial growth, which was observed in the field when the injection of neat toluene was employed . Over 90% removal of TCE was achieved with primary substrate (toluene) degraded to a concentration below 10 microg/L. J Cell Mol Med, 2004 Jul-Sep, 8(3), 317 - 28 Immunomodulatory benefits of cyclosporine A in inflammatory bowel disease; Kountouras J et al.; Etiopathogenesis of mucosal inflammation in inflammatory bowel disease remains a complex and enigmatic field; various factors (genetic, environmental and microbial) trigger an event that activates intestinal immune and nonimmune systems culminating in inflammation and tissue injury . Specifically, both innate and adaptive immune systems seem to play important roles in the pathophysiology of this disease . Cyclosporine A represents a macrolide immune modulator with primary inhibitory effects on T helper lymphocyte production of interleukin-2, and other cytokines leading to altered T-lymphocyte and B-lymphocyte function . The diversity of its therapeutic outcome reported in inflammatory bowel disease may be due to the intricate immuno-pathogenic profile of the disease and the variety of the applied dose-dependent courses of therapy . Cyclosporine A exerts additional actions on other components of the inflammatory infiltrate, including neutrophils and mast cells, thereby appearing to be a multi-dynamic therapeutic approach, although with potential drawbacks, that may be applied alone or combined with other immunomodulatory agents in inflammatory bowel disease patients . Because cyclosporine A induces apoptosis of T-lymphocytes responsible for perpetuation of the chronic inflammatory process in the disease with potential tumorigenic effect, it may exert a further inhibitory effect on cancer development in inflammatory bowel disease patients, and can be combined with other relative agents, such as rapamycin, which also promotes T-lymphocyte apoptosis . Therefore, recently established multifactorial action of cyclosporine A in relation to the pathogenesis of the disease can open new horizons for prospective, controlled trials in large cohorts, aiming to emphasize cyclosporine A's potential. J Periodontal Res, 2004 Dec, 39(6), 447 - 52 Factor XIIIa+ dendritic cells and S-100 protein+ Langerhans' cells in adult periodontitis; Dereka XE et al.; OBJECTIVE: The purpose of this study was to evaluate the presence of factor XIIIa+ dendritic cells and S-100 protein+ Langerhans' cells in the gingival epithelium and connective tissue of periodontal pockets, before and after non-surgical periodontal therapy . BACKGROUND: The microbial flora in periodontal pockets provokes complex immune reactions . Dendritic cells play a critical role in primary and secondary immune responses and are considered as antigen-presenting cells . Factor XIIIa positive dendritic cells and S-100 protein positive Langerhans' cells identified by immunoreactivity against factor XIIIa antigen and S-100 protein, respectively, are two distinct subpopulations of dendritic cells . METHODS: Fifty-four gingival tissue samples were obtained from periodontal pockets of initial depth 4-5 mm and > or = 6 mm . Each group was subdivided in to three subgroups . The first subgroup consisted of samples taken on baseline day and used as control . The second and third subgroups included those obtained 1 month after plaque and calculus removal, and 1 month after scaling and root planing, respectively, additionally to oral hygiene instructions . The tissues were removed from the palatal gingiva under local anaesthesia during routine periodontal surgery . Immunohistochemical staining with antibodies against factor XIIIa and S-100 protein was performed to identify dendritic cells positive and Langerhans' cells positive, respectively . RESULTS: Factor XIIIa+ dendritic cell numbers decreased compared to controls after plaque and calculus removal, oral hygiene instructions and scaling and root planing in periodontal pockets of 4-5 mm, but not in those of > or = 6 mm depth . S-100+ Langerhans' cell numbers decreased after periodontal treatment in the periodontal pockets > or = 6 mm . CONCLUSION: These results may reflect a tendency for reduction of these two distinctive subpopulations of dendritic cells after non-surgical periodontal therapy . (c)Blackwell Munksgaard 2004 J Biomed Mater Res B Appl Biomater, 2005 Jan 15, 72(1), 37 - 42 Enzymatic stabilization of gelatin-based scaffolds; Broderick EP et al.; The definitive goal of this research is to develop protein-based scaffolds for use in soft tissue regeneration, particularly in the field of dermal healing . The premise of this investigation was to characterize the mechanical properties of gelatin cross-linked with microbial transglutaminase (mTGase) and to investigate the cytocompatibility of mTGase cross-linked gelatin . Dynamic rheological analysis revealed a significant increase in the storage modulus and thermal stability of gelatin after cross-linking with mTGase . Static, unconfined compression tests showed an increase in Young's modulus of gelatin gels after mTGase cross-linking . A comparable increase in gel strength was observed with 0.03% mTGase and 0.25% glutaraldehyde cross-linked gelatin gels . In vitro studies using 3T3 fibroblasts indicated cytotoxicity at a concentration of 0.05% mTGase after 72 h . However, no significant inhibition of cell proliferation was seen with cells grown on lower concentrations of mTGase cross-linked gelatin substrates . The mechanical improvement and cytocompatibility of mTGase cross-linked gelatin suggests mTGase has potential for use in stabilizing gelatin gels for tissue-engineering applications . J Biol Chem, 2005 Jan 7, 280(1), 484 - 93 Epub 2004 Oct 15. The 1.8-a crystal structure of human tear lipocalin reveals an extended branched cavity with capacity for multiple ligands; Breustedt DA et al.; In contrast with earlier assumptions, which classified human tear lipocalin (Tlc) as an outlier member of the lipocalin protein family, the 1.8-A resolution crystal structure of the recombinant apoprotein confirms the typical eight-stranded antiparallel beta-barrel architecture with an alpha-helix attached to it . The fold of Tlc most closely resembles the bovine dander allergen Bos d 2, a well characterized prototypic lipocalin, but also reveals similarity with beta-lactoglobulin . However, compared with other lipocalin structures Tlc exhibits an extremely wide ligand pocket, whose entrance is formed by four partially disordered loops . The cavity deeply extends into the beta-barrel structure, where it ends in two distinct lobes . This unusual structural feature explains the known promiscuity of Tlc for various ligands, with chemical structures ranging from lipids and retinoids to the macrocyclic antibiotic rifampin and even to microbial siderophores . Notably, earlier findings of biological activity as a thiol protease inhibitor have no correspondence in the three-dimensional structure of Tlc, rather it appears that its proteolytic fragments could be responsible for this phenomenon . Hence, the present structural analysis sheds new light on the ligand binding activity of this functionally obscure but abundant human lipocalin. Chemosphere, 2004 Nov, 57(8), 853 - 62 Laboratory assessment of atrazine and fluometuron degradation in soils from a constructed wetland; Weaver MA et al.; Constructed wetlands offer promise for removal of nonpoint source contaminants such as herbicides from agricultural runoff . Laboratory studies assessed the potential of soils to degrade and sorb atrazine and fluometuron within a recently constructed wetland . The surface 3 cm of soil was sampled from two cells of a Mississippi Delta constructed wetland; one shallow area disturbed only hydrologically, and the second excavated to provide greater water-holding capacity . The excavated area was more acidic on average (pH 4.85 versus 5.21), but otherwise the physical properties and general microbial enzyme activities in the two areas were similar . Soils were treated with 84 and 68 microg kg(-1) soil (14)C-ring labeled atrazine and fluometuron, respectively, and incubated under either saturated (88% moisture, w:w) or flooded (1cm standing water) conditions . Soils were sampled over 32 days and extracted for herbicide and metabolite analysis . Under saturated conditions, fluometuron metabolized to desmethylfluometuron (DMF) with a half-life equal 25-27 days . However, under flooded conditions, the half-life of fluometuron was more than 175 days . Atrazine dissipated rapidly in saturated and flooded soil with a half-life of approximately 23 days, but only 10% of atrazine was mineralized to CO(2) . The overall atrazine and fluometuron dissipation rates were similar between the two cells, but each area had a different pattern of metabolite accumulation . The major route of atrazine dissipation was incorporation of atrazine residues into methanol-nonextractable (soil-bound) components, with minimal extractable metabolite accumulation . A mixed-mode extractant (potassium phosphate:acetonitrile) recovered greater amounts of (14)C-residues from atrazine-treated soils, suggesting that hydrolysis of atrazine to hydroxylated metabolites was a major component of the bound residues . These studies indicate the potential for herbicide dissipation in wetland soils and a differential effect of flooding on the fate of these herbicides. Annu Rev Microbiol, 2004, 58, 329 - 63 Johne's disease, inflammatory bowel disease, and Mycobacterium paratuberculosis; Chacon O et al.; Johne's disease is a chronic diarrhea affecting all ruminants . Mycobacterium avium subsp . paratuberculosis (MAP), a slowly growing mycobacteria, is the etiologic agent . There is also a concern that MAP might be a causative agent of some cases of inflammatory bowel disease in humans, especially Crohn's disease . Food products including pasteurized bovine milk have been suggested as potential sources of human infection . This review addresses microbial factors that may contribute to its pathogenicity . In addition, the experimental evidence defining MAP as the cause of Johne's disease and the issues and controversies surrounding its potential pathogenic role in humans are discussed. Annu Rev Microbiol, 2004, 58, 1 - 42 Pickles, pectin, and penicillin; Demain AL; My professional life has been devoted to the study of microbial products and their biosynthesis, regulation, and overproduction . These have included primary metabolites (glutamic acid, tryptophan, inosinic acid, guanylic acid, vitamin B(12), riboflavin, pantothenic acid, ethanol, and lactic acid) and secondary metabolites (penicillin, cephalosporins, streptomycin, fosfomycin, gramicidin S, rapamycin, indolmycin, microcin B17, fumagillin, mycotoxins, Monascus pigments, and tetramethylpyrazine) . Other areas included microbial nutrition, strain improvement, bioconversions of statins and beta-lactams, sporulation and germination, plasmid stability, gel microdroplets, and the production of double-stranded RNA, the polymer xanthan, and enzymes (polygalacturonase, protease, cellulase) . Most of the studies were carried out with me by devoted and hardworking industrial scientists for 15 years at Merck & Co . and by similarly characterized students, postdoctorals, and visiting scientists during my 32 years at the Massachusetts Institute of Technology . I owe much of my success to my mentors from academia and industry . My recent research activities with undergraduate students at the Charles A . Dana Research Institute for Scientists Emeriti (R.I.S.E.) at Drew University have been very rewarding and are allowing me to continue my career. Environ Sci Technol, 2004 Sep 15, 38(18), 4723 - 7 Effect of storage on the isotopic composition of nitrate in bulk precipitation; Spoelstra J et al.; Stable isotopic analysis of atmospheric nitrate is increasingly employed to study nitrate sources and transformations in forested catchments . Large volumes have typically been required for delta18O and delta15N analysis of nitrate in precipitation due to relatively low nitrate concentrations . Having bulk collectors accumulate precipitation over an extended time period allows for collection of the required volume as well as reducing the total number of analyses needed to determine the isotopic composition of mean annual nitrate deposition . However, unfiltered precipitation left in collectors might be subject to microbial reactions that can alter the isotopic signature of nitrate in the sample . Precipitation obtained from the Turkey Lakes Watershed was incubated under conditions designed to mimic unfiltered storage in bulk precipitation collectors and monitored for changes in nitrate concentration, delta15N, and delta18O . Results of this experiment indicated that no detectable nitrate production or assimilation occurred in the samples during a two-week incubation period and that atmospheric nitrate isotopic ratios were preserved . The ability to collect unfiltered precipitation samples for an extended duration without alteration of nitrate isotope ratios is particularly useful at remote study sites where daily retrieval of samples may not be feasible. Vet Hum Toxicol, 2004 Oct, 46(5), 248 - 50 Safety of Pochonia chlamydosporia var catenulata in acute oral and dermal toxicity/pathogenicity evaluations in rats and rabbits; Garcia L et al.; The nematophagous fungus, Pochonia chlamydosporia var . catenulata (Kamyschlco ex Barron & Onions) Zare & W-Gams, was investigated as a potential biocontrol agent in integrated pest management strategy for Meloidogyne incognita (Kofoid and White) Chitwood in vegetable crops in Cuba . An acute oral and dermal toxicity/patogenicity study was performed to determine the safety of this fungus in non-target organisms . In the first study, a 1-dose level of 5 x 10(8) units of the microbial pest control agent/treated rat was used . Mortality or clinical signs were not evident and no adverse effects on body weight, hematology, microbiology and gross or microscopic pathology were observed . Food and water consumption was not significantly different between control and treated groups . In the acute dermal toxicity study, there was neither mortality nor clinical signs of toxicity, and no toxic effects in gross and microscopic pathology were detected . Thus, Pochonia chlamydosporia var . catenulate (Vcc-108, IMI SD 187), administered oral and dermally to rats and rabbits respectively, was safe in toxicity/pathogenicity studies. J Biol Chem, 2004 Dec 24, 279(52), 54405 - 15 Epub 2004 Oct 13. Stimulation of toll-like receptor 2 by Coxiella burnetii is required for macrophage production of pro-inflammatory cytokines and resistance to infection; Zamboni DS et al.; Innate and adaptive immune responses are initiated upon recognition of microbial molecules by Toll-like receptors (TLRs) . We have investigated the importance of these receptors in the induction of pro-inflammatory cytokines and macrophage resistance to infection with Coxiella burnetii, an obligate intracellular bacterium and the etiological agent of Q fever . By using a Chinese hamster ovary/CD14 cell line expressing either functional TLR2 or TLR4, we determined that C . burnetii phase II activates TLR2 but not TLR4 . Macrophages deficient for TLR2, but not TLR4, produced less tumor necrosis factor-alpha and interleukin-12 upon C . burnetii infection . Furthermore, it was found that TLR2 activation interfered with C . burnetii intracellular replication, as macrophages from TLR2-deficient mice were highly permissive for C . burnetii growth compared with macrophages from wild type mice or TLR4-deficient mice . Although LPS modifications distinguish virulent C . burnetii phase I bacteria from avirulent phase II organisms, electrospray ionization-mass spectrometry analysis showed that the lipid A moieties isolated from these two phase variants are identical . Purified lipid A derived from either phase I or phase II LPS failed to activate TLR2 and TLR4 . Indeed, the lipid A molecules were able to interfere with TLR4 signaling in response to purified Escherichia coli LPS . These studies indicate that TLR2 is an important host determinant that mediates recognition of C . burnetii and a response that limits growth of this intracellular pathogen. Water Sci Technol, 2004, 50(4), 233 - 40 Volatile organic compound adsorption in a gas-solid fluidized bed; Ng YL et al.; Fluidization finds many process applications in the areas of catalytic reactions, drying, coating, combustion, gasification and microbial culturing . This work aims to compare the dynamic adsorption characteristics and adsorption rates in a bubbling fluidized bed and a fixed bed at the same gas flow-rate, gas residence time and bed height . Adsorption with 520 ppm methanol and 489 ppm isobutane by the ZSM-5 zeolite of different particle size in the two beds enabled the differentiation of the adsorption characteristics and rates due to bed type, intraparticle mass transfer and adsorbate-adsorbent interaction . Adsorption of isobutane by the more commonly used activated carbon provided the comparison of adsorption between the two adsorbent types . With the same gas residence time of 0.79 seconds in both the bubbling bed and fixed bed of the same bed size of 40 mm diameter and 48 mm height, the experimental results showed a higher rate of adsorption in the bubbling bed as compared to the fixed bed . Intraparticle mass transfer and adsorbent-adsorbate interaction played significant roles in affecting the rate of adsorption, with intraparticle mass transfer being more dominant . The bubbling bed was observed to have a steeper decline in adsorption rate with respect to increasing outlet concentration compared to the fixed bed . The adsorption capacities of zeolite for the adsorbates studied were comparatively similar in both beds; fluidizing, and using smaller particles in the bubbling bed did not increase the adsorption capacity of the ZSM-5 zeolite . The adsorption capacity of activated carbon for isobutane was much higher than the ZSM-5 zeolite for isobutane, although at a lower adsorption rate . Fourier transform infra-red (FTIR) spectroscopy was used as an analytical tool for the quantification of gas concentration . Calibration was done using a series of standards prepared by in situ dilution with nitrogen gas, based on the ideal gas law and relating partial pressure to gas concentration . Concentrations up to 220 ppm for methanol and 75 ppm for isobutane were prepared using this method. Am J Physiol Cell Physiol, 2005 Feb, 288(2), C321 - 8 Epub 2004 Oct 13. Chemokine receptor CCR6 transduces signals that activate p130Cas and alter cAMP-stimulated ion transport in human intestinal epithelial cells; Yang CC et al.; Human colon epithelial cells express the G protein-coupled receptor CCR6, the sole receptor for the chemokine CCL20 (also termed MIP-3alpha) . CCL20 produced by intestinal epithelial cells is upregulated in response to proinflammatory stimuli and microbial infection, and it chemoattracts leukocytes, including CCR6-expressing immature myeloid dendritic cells, into sites of inflammation . The aim of this study was to determine whether CCR6 expressed by intestinal epithelial cells acts as a functional receptor for CCL20 and whether stimulation with CCL20 alters intestinal epithelial cell functions . The human colon epithelial cell lines T84, Caco-2, HT-29, and HCA-7 were used to model colonic epithelium . Polarized intestinal epithelial cells constitutively expressed CCR6, predominantly on the apical side . Consistent with this, apical stimulation of polarized intestinal epithelial cells resulted in tyrosine phosphorylation of the p130 Crk-associated substrate (Cas), an adaptor/scaffolding protein that localizes in focal adhesions and has a role in regulating cytoskeletal elements important for cell attachment and migration . In addition, CCL20 stimulation inhibited agonist-stimulated production of the second messenger cAMP and cAMP-mediated chloride secretory responses by intestinal epithelial cells . Inhibition was abrogated by pertussis toxin, consistent with signaling through Galpha(i) proteins that negatively regulate adenylyl cyclases and cAMP production . These data indicate that signaling events, occurring via the activation of the apically expressed chemokine receptor CCR6 on polarized intestinal epithelial cells, alter specialized intestinal epithelial cell functions, including electrogenic ion secretion and possibly epithelial cell adhesion and migration. Expert Rev Anti Infect Ther, 2003 Aug, 1(2), 231 - 9 Pneumolysin in the immunopathogenesis and treatment of pneumococcal disease; Cockeran R et al.; Recent insights into the immunopathogenesis of pneumococcal infection, a common and significant cause of morbidity and mortality, have implicated pneumolysin as being a prominent virulence factor, which may play a role in microbial colonization, invasion and dissemination, as well as tissue inflammation . Being a highly immunogenic polypeptide produced by all clinically relevant pneumococcal isolates, pneumolysin is recognized as a potential carrier protein for polysaccharide conjugate vaccines, while in the setting of acute disease, promising pneumolysin-directed pharmacological strategies include, among others, macrolides and corticosteroids. Expert Rev Anti Infect Ther, 2003 Jun, 1(1), 167 - 73 The art of serendipity: killing of Caenorhabditis elegans by human pathogens as a model of bacterial and fungal pathogenesis; Mylonakis E et al.; The nematode worm, Caenorhabditis elegans, has been used to develop a facile model system of host-pathogen interactions to identify basic evolutionarily conserved pathways associated with microbial pathogenesis . The model involves the killing of Caenorhabditis elegans by a variety of human pathogens . Several virulence-related genes in a variety of pathogens previously shown to be involved in mammalian infection have also been shown to play a role in Caenorhabditis elegans killing . Screening of large numbers of microbial mutants for attenuation in a mammalian model would require thousands of mice, rats or rabbits . In contrast, the Caenorhabditis elegans model allows rapid identification of mutants in microbial genes associated with pathogenesis and then these phenotypes can be confirmed in a relevant mammalian model. Pest Manag Sci, 2004 Oct, 60(10), 1025 - 34 Integrated pest management approach for a new pest, Lacanobia subjuncta (Lepidoptera: Noctuidae), in Washington apple orchards; Doerr MD et al.; Bioassays of Lacanobia subjuncta (Grote and Robinson) larvae established baseline LC50 values and identified the potential of reduced-risk, organophosphate replacement and naturally derived insecticides (eg chloronicotinyls, spinosyns, oxadiazines, insect growth regulators, microbial insecticides and particle films) to control this pest . The toxicities of these products were compared with those of organophosphate, carbamate, chlorinated cyclodiene and synthetic pyrethroid insecticides used in the management of lepidopteran pests in Washington apple orchards . Field trials were conducted comparing candidate insecticides to conventional alternatives . Several new insecticides (eg spinosad, methoxyfenozide, indoxacarb and an aluminosilicate particle film) proved to be effective for the management of L subjuncta . We summarize the goals and challenges of developing an integrated pest management program for new and resurgent pests as insecticide tools continue to change, and propose a hypothesis for the sudden increase in pest status of L subjuncta based on organophosphate tolerances . The role of novel insecticides with unique modes of action in resistance management and the encouragement of biological control are also discussed. Immunobiology, 2004, 209(1-2), 191 - 8 Heterogeneity of TLR-induced responses in dendritic cells: from innate to adaptive immunity; Re F et al.; Toll-like receptors (TLR) mediate recognition of several microbial products . Accumulating evidence indicates that TLR are capable of inducing distinct responses in dendritic cells and other antigen-presenting cells, and can direct T-helper cell differentiation in opposing directions . The generation of such varied responses is achieved through the selective utilization of adaptor molecules that link TLR to distinct signal transduction pathways . The ability of TLR to activate and guide innate and adaptive immunity has the potential to be exploited for practical application that may lead to the development of more successful immunotherapies and vaccination strategies . A review of recent literature, unpublished observations, and future challenges is presented here. Immunobiology, 2004, 209(1-2), 51 - 6 The role of matrix metalloproteinase 7 in innate immunity; Burke B; Matrix metalloproteinase 7 (MMP-7), or matrilysin, is a secreted protease expressed by glandular and mucosal epithelial cells, keratinocytes, fibroblasts and macrophages . As with other MMPs it can act on the extracellular matrix and thereby regulate cell migration and tissue repair . In addition, MMP-7 has an important role in the maintenance of innate immunity in organs such as the lungs and intestines where it proteolytically activates anti-bacterial peptides such as pro-defensins . MMP-7 is also important for mediating proteolytic release of TNF from macrophages . Consistent with its role in innate immunity, MMP-7 is induced by microbial products and also, unexpectedly, by hypoxia. Immunobiology, 2004, 209(1-2), 39 - 49 The role of scavenger receptors in pathogen recognition and innate immunity; Mukhopadhyay S et al.; Scavenger receptors represent a large family of structurally unrelated distinct gene products, expressed by myeloid and selected endothelial cells and able to recognise modified low-density lipoproteins . They also bind and internalise a variety of microbial pathogens, as well as modified or endogenous molecules derived from the host, and contribute to a range of physiological or pathological processes. J Cell Physiol, 2005 Mar, 202(3), 849 - 51 Amniotic membrane transplantation in ocular surface disorders; Tosi GM et al.; Chronic ocular surface disorders, which can result in severe functional impairment, have been viewed for decades as untreatable diseases . In 1995, the reintroduction of amniotic membrane transplantation (AMT), either alone or associated with limbal stem cell transplantation, has offered new hope of using tissue and cell therapy strategies to repair ocular surface disorders . Amniotic membrane (AM) has been found to exert its effects by acting as a substrate for the growth of ocular surface epithelia, by suppressing inflammation and scarring and by serving as an anti-microbial barrier . Moreover, AM has recently been used as a substrate for ex vivo expansion of corneal epithelial cells for ocular surface reconstruction . Notwithstanding the substantial agreement among Authors regarding its clinical efficacy, there are still many uncertainties regarding the fate of grafted AM and consequently the mechanisms through which it exerts its long-term effects . Further studies including controlled clinical trials with numerous cases are required to understand which ocular surface conditions are certain to benefit from AM transplantation and how its mechanical properties interact with the mediators produced to favor ocular surface reconstruction . (c) 2004 Wiley-Liss, Inc. Appl Microbiol Biotechnol, 2004 Sep, 65(4), 488 - 95 Epub 2004 Jul 20. Microbial population dynamics during fed-batch operation of commercially available garbage composters; Narihiro T et al.; Microbial populations in terms of quantity, quality, and activity were monitored during 2 months of start-up operation of commercially available composters for fed-batch treatment of household biowaste . All the reactors, operated at a waste-loading rate of 0.7 kg day(-1) (wet wt), showed a mass reduction efficiency of 88-93% . The core temperature in the reactors fluctuated between 31 degrees C and 58 degrees C due to self-heating . The pH declined during the early stage of operation and steadied at pH 7.4-9.3 during the fully acclimated stage . The moisture content was 48-63% early in the process and 30-40% at the steady state . Both direct total counts and plate counts of bacteria increased via two phases (designated phases I, II) and reached an order of magnitude of 10(11) cells g(-1) (dry wt) at the steady state . Microbial community changes during the start-up period were studied by culture-independent quinone profiling and denatured gradient gel electrophoresis (DGGE) of PCR-amplified 16S rDNA . In all the reactors, ubiquinones predominated during phase I, whereas partially saturated menaquinones became predominant during phase II . This suggested that there was a drastic population shift from ubiquinone-containing Proteobacteria to Actinobacteria during the start-up period . The DGGE analysis of the bacterial community in one of the reactors also demonstrated a drastic population shift during phase I and the predominance of members of the phyla Proteobacteria and Bacteroidetes during the overall period . But this molecular analysis failed to detect actinobacterial clones from the reactor at any stage. Environ Microbiol, 2004 Nov, 6(11), 1159 - 73 Acetoclastic and hydrogenotrophic methane production and methanogenic populations in an acidic West-Siberian peat bog; Kotsyurbenko OR et al.; Sites in the West Siberian peat bog 'Bakchar' were acidic (pH 4.2-4.8), low in nutrients, and emitted CH4 at rates of 0.2-1.5 mmol m(-2) h(-1) . The vertical profile of delta13CH4 and delta13CO2 dissolved in the porewater indicated increasing isotope fractionation and thus increasing contribution of H2/CO2-dependent methanogenesis with depth . The anaerobic microbial community at 30-50 cm below the water table produced CH4 with optimum activity at 20-25 degrees C and pH 5.0-5.5 respectively . Inhibition of methanogenesis with 2-bromo-ethane sulphonate showed that acetate, phenyl acetate, phenyl propionate and caproate were important intermediates in the degradation pathway of organic matter to CH4 . Further degradation of these intermediates indicated that 62-72% of the CH4 was ultimately derived from acetate, the remainder from H2/CO2 . Turnover times of {2-14C}acetate were on the order of 2 days (15, 25 degrees C) and accounted for 60-65% of total CH4 production . Conversion of 14CO2 to 14CH4 accounted for 35-43% of total CH4 production . These results showed that acetoclastic and hydrogenotrophic methanogenesis operated closely at a ratio of approximately 2 : 1 irrespective of the incubation temperature (4, 15 and 25 degrees C) . The composition of the archaeal community was determined in the peat samples by terminal restriction fragment length polymorphism (T-RFLP) analysis and sequencing of amplified SSU rRNA gene fragments, and showed that members of Methanomicrobiaceae, Methanosarcinaceae and Rice cluster II (RC-II) were present . Other, presumably non-methanogenic archaeal clusters (group III, RC-IV, RC-V, RC-VI) were also detected . Fluorescent in situ hybridization (FISH) showed that the number of Bacteria decreased (from 24 x 10(7) to 4 x 10(7) cells per gram peat) with depth (from 5 to 55 cm below the water table), whereas the numbers of Archaea slightly increased (from 1 x 10(7) to 2 x 10(7) cells per gram peat) . Methanosarcina spp . accounted for about half of the archaeal cells . Our results show that both hydrogenotrophic and acetoclastic methanogenesis are an integral part of the CH4-producing pathway in acidic peat and were represented by appropriate methanogenic populations. Environ Microbiol, 2004 Nov, 6(11), 1123 - 36 Streptomycin as a selective agent to facilitate recovery and isolation of introduced and indigenous Sphingomonas from environmental samples; Vanbroekhoven K et al.; Sphingomonas is an organism of major interest for the degradation of organic contaminants in soils and other environments . A medium based on the aminoglycoside antibiotic streptomycin (Sm) was developed, which, together with the yellow pigmentation of Sphingomonas, facilitated the detection, recovery and quantification of culturable Sphingomonas from soils . All 29 previously described bacterial strains belonging to 17 different Sphingomonas species were able to grow on mineral media containing 200 microg ml(-1) streptomycin, showing that the capacity to resist high concentrations of Sm is a common characteristic within Sphingomonas . Incorporation of Sm into the mineral medium led to a significant reduction in the background microbial population and a concomitant 100 times more sensitive detection of Sphingomonas inoculated in non-sterile soil matrices . The Sm-containing medium was used to examine a variety of hydrocarbon-contaminated soils for the presence and biodiversity of Sphingomonas . Incorporation of Sm in the medium led to a significant increase in the number of yellow-pigmented colonies . Comparison of contaminated and non-contaminated soils derived from the same site revealed colonization by culturable yellow-pigmented Sm-resistant bacteria of the polluted location solely . Both yellow and non-yellow-pigmented colonies were purified from plates containing glucose and Sm, and BOX-polymerase chain reaction (PCR) was used to sort out clonally related strains . Representative strains from the major BOX-PCR clusters were identified using FAME and partial 16S rRNA gene sequencing . Forty-eight of 58 Sm-resistant isolates were identified as Sphingomonas sp . Streptomycin-resistant Sphingomonas isolates generated BOX-PCR diversity patterns that were site dependent and represented different species mainly belonging to Sphingomonas subgroups containing species formerly designated as Sphingopyxis and Sphingobium . The ability to degrade phenanthrene was only found in a minority of the Sphingomonas isolates, which all originated from soils containing high phenanthrene concentrations. Environ Microbiol, 2004 Nov, 6(11), 1109 - 22 Bacterial endosymbionts of insects: insights from comparative genomics; Gil R et al.; The development of molecular techniques for the study of uncultured bacteria allowed the extensive study of the widespread association between insects and intracellular symbiotic bacteria . Most of the bacterial endosymbionts involved in such associations are gamma-proteobacteria, closely related to Escherichia coli . In recent years, five genomes from insect endosymbionts have been sequenced, allowing the performance of extensive genome comparative analysis that, as a complement of phylogenetic studies, and analysis on individual genes, can help to understand the different traits of this particular association, including how the symbiotic process is established, the explanation of the special features of these microbial genomes, the bases of this intimate association and the possible future that awaits the endosymbionts with extremely reduced genomes. Keio J Med, 2004 Sep, 53(3), 151 - 8 Peripheral B lymphocyte tolerance; Gavin A et al.; This lecture discusses two interrelated topics, B cell tolerance in the peripheral immune system and BAFF . Using the 3-83 antibody transgenic mouse bred to mice carrying cognate antigen in the liver, we previously found that clonal elimination drastically reduced the precursor frequency of autoreactive cells . The consensus model to explain this tolerance is the 2-signal hypothesis, which proposes that in the absence of T cell help BCR stimulation is a negative signal for B cells . However, this model fails to explain how these same B cells can respond to T-independent type II (TI-2) antigens, raising the question of how they distinguish TI-2 antigens from multimeric self determinants . We propose that B cells use NK-like missing self recognition to provide the needed specificity, as foreign antigens are unlikely to carry self markers . The model has implications for the evolution of the immune system, B lymphocyte signaling, tissue specificity of autoimmunity, and microbial subversion of the immune system . Overexpression of the critical B cell survival cytokine BAFF/BLyS has been associated with autoimmunity . We have discovered a novel splice isoform that regulates BAFF activity and may play a role in limiting B cell activity . The novel form, called DBAFF, is able to heteromultimerize with normal BAFF and can suppress receptor binding and proteolytic release from the cell surface . Preliminary studies from transgenic mice overexpressing wild type or DBAFF are consistent with a possible regulatory role for DBAFF, raising the possibility that the relative expression levels of BAFF and DBAFF regulates tolerance. Mar Pollut Bull, 2004 Oct, 49(7-8), 537 - 47 Ecosystem history of Mississippi River-influenced continental shelf revealed through preserved phytoplankton pigments; Rabalais NN et al.; Pigments determined by high performance liquid chromatography (HPLC) provide useful information concerning water column and epibenthic plant and microbial communities in both extant communities and accumulated sediments in lakes, estuaries and the ocean . Chlorophyll and its degradation products provide an estimate of overall biomass, and carotenoid pigments provide taxonomic biomarkers of phytoplankton . We examined the pigments preserved in sediment cores from the Louisiana continental shelf adjacent to the outflow of the Mississippi River system to document changes in phytoplankton community composition, phytoplankton abundance, and conditions of hypoxia over time . Carbon accumulated in sediments from water depths of 20-60 m is primarily derived from marine phytoplankton and represents the history of phytoplankton communities in the overlying water . There is a general increase in chlorophyll a, pheopigments, zeaxanthin, fucoxanthin and most carotenoids over time, with the change gradual from 1955 to 1970, followed by a fairly steady increase to 1997 . The highest chloropigment concentrations are in cores from areas more likely to be exposed to seasonal hypoxia . These indicate an increase in eutrophication in the form of greater diatom and cyanobacterial production, or a worsening of hypoxia, or both . This trend expanded westward along the Louisiana shelf in the 1990s. BMC Genomics . 2004 Oct 09;5(1):78. i-Genome: a database to summarize oligonucleotide data in genomes; Lin FM et al.; BACKGROUND: Information on the occurrence of sequence features in genomes is crucial to comparative genomics, evolutionary analysis, the analyses of regulatory sequences and the quantitative evaluation of sequences . Computing the frequencies and the occurrences of a pattern in complete genomes is time-consuming . RESULTS: The proposed database provides information about sequence features generated by exhaustively computing the sequences of the complete genome . The repetitive elements in the eukaryotic genomes, such as LINEs, SINEs, Alu and LTR, are obtained from Repbase . The database supports various complete genomes including human, yeast, worm, and 128 microbial genomes . CONCLUSIONS: This investigation presents and implements an efficiently computational approach to accumulate the occurrences of the oligonucleotides or patterns in complete genomes . A database is established to maintain the information of the sequence features, including the distributions of oligonucleotide, the gene distribution, the distribution of repetitive elements in genomes and the occurrences of the oligonucleotides . The database can provide more effective and efficient way to access the repetitive features in genomes. J Environ Sci Health B, 2004 May, 39(4), 517 - 31 Dissipation of chlorpyrifos in two soil environments of semi-arid India; Menon P et al.; The dissipation of chlorpyrifos (20 EC) at environment-friendly doses in the sandy loam and loamy sand soils of two semi-arid fields and the presence of pesticide residues in the harvested groundnut seeds, were monitored . The movement of chlorpyrifos through soil and its binding in the loamy sand soil was studied using 14C chlorpyrifos . Chlorpyrifos was moderately stable in both loamy sand and sandy loam soils, with half-life of 12.3 and 16.4 days, respectively . With 20 EC treatments the dissipation was slower for standing crop than seed treatment, indicative of the high degradation rates in the bioactive rhizosphere . In soil, 3,5,6-trichloro-2-pyridinol (TCP) was the principal breakdown product . Presence of 3,5,6-trichloro-2-methoxypyridine (TMP), the secondary metabolite, detected in the rhizospheric samples during this study, has not been reported earlier in field soils . The rapid dissipation of the insecticide from the soil post-application might have resulted from low sorption due to the alkalinity of the soil and its low organic matter content, fast topsoil dissipation possibly by volatilization and photochemical degradation, aided by the low water solubility, limited vertical mobility due to confinement of residues to the upper 15 cm soil layers and microbial mineralization and nucleophilic hydrolysis . Contrary to the reports of relatively greater mobility of its metabolites in temperate soils, TMP and TCP remained confined to the top 15 cm soil . The formation of bound residues (half-life 13.4 days) in the loamy sand soil was little and not "irreversible." A decline in bound residues could be correlated to decreasing TCP concentration . Higher pod yields were obtained from pesticide treated soils in comparison to controls . Post-harvest no pesticide residues were detected in the soils and groundnut seeds. Proteomics, 2004 Oct, 4(10), 2829 - 2830 Proteomics of microbial pathogens; Jungblut PR et al.; No abstracts. J Anim Sci, 2004, 82 E-Suppl, E83 - 92 Building beef cow nutritional programs with the 1996 NRC beef cattle requirements model; Lardy GP et al.; Designing a sound cow-calf nutritional program requires knowledge of nutrient requirements, diet quality, and intake . Effectively using the NRC (1996) beef cattle requirements model (1996NRC) also requires knowledge of dietary degradable intake protein (DIP) and microbial efficiency . Objectives of this paper are to 1) describe a framework in which 1996NRC-applicable data can be generated, 2) describe seasonal changes in nutrients on native range, 3) use the 1996NRC to predict nutrient balance for cattle grazing these forages, and 4) make recommendations for using the 1996NRC for forage-fed cattle . Extrusa samples were collected over 2 yr on native upland range and subirrigated meadow in the Nebraska Sandhills . Samples were analyzed for CP, in vitro OM digestibility (IVOMD), and DIP . Regression equations to predict nutrients were developed from these data . The 1996NRC was used to predict nutrient balances based on the dietary nutrient analyses . Recommendations for model users were also developed . On subirrigated meadow, CP and IVOMD increased rapidly during March and April . On native range, CP and IVOMD increased from April through June but decreased rapidly from August through September . Degradable intake protein (DM basis) followed trends similar to CP for both native range and subirrigated meadow . Predicted nutrient balances for spring- and summer-calving cows agreed with reported values in the literature, provided that IVOMD values were converted to DE before use in the model (1.07 x IVOMD - 8.13) . When the IVOMD-to-DE conversion was not used, the model gave unrealistically high NE(m) balances . To effectively use the 1996NRC to estimate protein requirements, users should focus on three key estimates: DIP, microbial efficiency, and TDN intake . Consequently, efforts should be focused on adequately describing seasonal changes in forage nutrient content . In order to increase use of the 1996NRC, research is needed in the following areas: 1) cost-effective and accurate commercial laboratory procedures to estimate DIP, 2) reliable estimates or indicators of microbial efficiency for various forage types and qualities, 3) improved estimates of dietary TDN for forage-based diets, 4) validation work to improve estimates of DIP and MP requirements, and 5) incorporation of nitrogen recycling estimates. Oper Dent, 2004 Sep-Oct, 29(5), 524 - 31 Fluoride releasing materials: their anti-cariogenic properties tested in in vitro caries models; Sa LT et al.; The anti-cariogenic properties of three fluoride-releasing materials on root surfaces were evaluated using two different caries models . Standardized cavities were prepared in dentin specimens and restored with either glass-ionomer, resin-modified glass-ionomer, polyacid-modified resin composite or resin composite material . Two groups of 56 specimens were demineralized using a microbial caries model for three days, while another 56 specimens were demineralized using a chemical demineralization model for three days . Lesions around the restorations were measured with a confocal laser scanning microscope . Glass ionomers demonstrated significant anti-cariogenic properties when exposed to the chemical model . However, no significant anti-cariogenic properties were observed with the microbial caries model . In conclusion, the fluoride releasing materials showed different anti-cariogenic properties in root surfaces under the different caries models, suggesting that caution should be exercised when trying to extrapolate the results of in vitro studies to the clinical situation. Indian J Med Sci, 2004 Sep, 58(9), 394 - 9 Microbial and cytopathological study of intrauterine contraceptive device users; Agarwal K et al.; CONTEXT: Intrauterine contraceptive device (IUCD) is a commonly used birth-spacing method which is fitted into maternal system . Clinical, microbial and cytopathological monitoring of women using these devices are important for ascertaining their side effects, risk of genital tract infection and carcinogenic potential . AIMS: To study clinical, microbial and cytopathological changes in IUCD users in a tertiary care hospital . DESIGN: Prospective analytic . SETTING: Tertiary hospital . MATERIAL AND METHODS: women visiting Family Planning clinic for follow up (IUCD users, n=100) or for IUCD insertion (controls, n=50) were enrolled in the study . Each subject underwent detailed history, general physical, systemic, and per local examination . Vaginal discharge was subjected to pH testing, KOH and wet mount examination, gram staining, and for culture and sensitivity . Bacterial vaginosis was defined using Nugent criteria . Cervical smears were examined and reported as per Bethesda system . STATISTICAL ANALYSIS: The information was entered into Microsoft Excel spreadsheet . The results were analyzed using EPI Info version-6 and Stata statistical software version 7 packages . Two-tailed t-test, chi2 test with Yates correction and two-tailed Fisher Exact tests were applied . RESULTS: Most women used CuT 200 (92%) . Median duration of use was 2 years . Chief complaints of IUCD users included backache (54%), vaginal discharge (46%), pain lower abdomen (34%), dyspareunia (22%), menorrhagia (18%) and dysmenorrhea (14%) . Mean hemoglobin was lower in IUCD users than controls (11.2+/-1.7 versus 11.9+/-1.8 g/dL, p 0.02) . Proportion of women with anemia was higher in IUCD users than in controls (29% versus 16%, p 0.12) . Cervical erosion was significantly increased in study group as compared the controls (20% versus 0%, p=0.00) whereas only insignificant increase in vaginitis (6% versus 0%, p=0.17) . Trichomonas vaginalis and fungal hyphae positivity and gram stain findings and bacterial vaginosis rate were not significantly different in two groups . Vaginal discharge bacterial culture revealed comparable results in two groups . Cytological findings were predominantly inflammatory . None of cases revealed Actinomycosis infection . High-grade squamous intraepithelial lesion (n=2) and low grade squamous intraepithelial lesion (n=1) cytological finding were present in IUCD users compared to none in controls . None of the cases had any malignant transformation . CONCLUSION: IUCD use results in lower hemoglobin concentration and cervical erosion . Women using IUCD requires a regular follow up, clinical examination, counseling and further investigation if required. J Immunol, 2004 Oct 15, 173(8), 5130 - 7 Bacterial heat shock proteins enhance class II MHC antigen processing and presentation of chaperoned peptides to CD4+ T cells; Tobian AA et al.; APCs process heat shock protein (HSP):peptide complexes to present HSP-chaperoned peptides on class I MHC molecules, but the ability of HSPs to contribute chaperoned peptides for class II MHC (MHC-II) Ag processing and presentation is unclear . Our studies revealed that exogenous bacterial HSPs (Escherichia coli DnaK and Mycobacterium tuberculosis HSP70) delivered an extended OVA peptide for processing and MHC-II presentation, as detected by T hybridoma cells . Bacterial HSPs enhanced MHC-II presentation only if peptide was complexed to the HSP, suggesting that the key HSP function was enhanced delivery or processing of chaperoned peptide Ag rather than generalized enhancement of APC function . HSP-enhanced processing was intact in MyD88 knockout cells, which lack most TLR signaling, further suggesting the effect was not due to TLR-induced induction of accessory molecules . Bacterial HSPs enhanced uptake of peptide, which may contribute to increased MHC-II presentation . In addition, HSPs enhanced binding of peptide to MHC-II molecules at pH 5.0 (the pH of vacuolar compartments), but not at pH 7.4, indicating another mechanism for enhancement of MHC-II Ag processing . Bacterial HSPs are a potential source of microbial peptide Ags during phagocytic processing of bacteria during infection and could potentially be incorporated in vaccines to enhance presentation of peptides to CD4+ T cells. Med J Malaysia, 2004 May, 59 Suppl B, 55 - 6 Microbial polyhydroxyalkanoates (PHAs): an emerging biomaterial for tissue engineering and therapeutic applications; Sudesh K; Among the various biomaterials available for tissue engineering and therapeutic applications, microbial polyhydroxyalkanoates (PHAs) offer the most diverse range of thermal and mechanical properties . Of particular interest are the PHAs that contain 4-hydroxybutyrate such as poly(3-hydroxybutyrate-co-4-hydroxybutyrate) {P(3HB-co-4HB) and poly(4-hydroxybutyrate) {P(4HB)} . These polyesters can only be synthesized by a few types of bacteria, among which Comamonas acidovorans has the most efficient metabolic pathways to channel 4HB monomers . The resulting polyesters are bioabsorbable and are being developed as a new biomaterial for medical applications . By controlling the molar ratio of the monomers, it is possible to produce materials that are as tough and elastic as rubber. Anesteziol Reanimatol, 2004 Jul-Aug, (4), 63 - 6 {Early nutrition support for intestinal insufficiency syndrome in spine surgery}; Molecular characterization of a novel splice site mutation within the CYBB gene leading to X-linked chronic granulomatous disease; Division of Immunology, Hospital de Ninos "Dr . Ricardo Gutierrez", Associated Hospital to the Universidad de Buenos Aires, Buenos Aires, ArgentinaChronic granulomatous disease (CGD) is a primary immunodeficiency that affects the oxidative mechanism of microbial killing of phagocytic cells . The defect is characterized by a lack or severely reduced superoxide anion (O(2) (-)) production by phagocytes . Seventy percent of CGD cases are X-linked (X-CGD) and they are caused by mutations in the gene encoding for gp91(phox), one of the two subunits of the flavocytochrome b558 of the NADPH oxidase . We identified an abnormal transcript arising from a novel splice site mutation within the gene encoding gp91(phox), which suggested that the mutation affected normal mRNA splicing . Thus, the effect of this mutation leads to the complete absence of the flavocytochrome b558 in neutrophil membranes, which caused the biochemical phenotype X91 degrees -CGD in this family . These molecular findings help to explain the early onset and severe phenotype in this X-CGD kindred . (c) 2004 Wiley-Liss, Inc. Macromol Biosci, 2004 Mar 15, 4(3), 330 - 9 Enzymatic hydrolysis of alpha- and beta-oligo(L-aspartic acid)s by poly(aspartic acid) hydrolases-1 and 2 from Sphingomonas sp . KT-1; Hiraishi T et al.; The enzymatic hydrolysis of alpha- and beta-oligo(L-aspartic acid)s by PAA hydrolase-1 and PAA hydrolase-2 (purified from Sphingomonas sp . KT-1) was performed to elucidate the mechanism of the microbial degradation by Sphingomonas sp . KT-1 of the thermally synthesized alpha,beta-poly(D,L-aspartic acid) (tPAA) . GPC analysis of the hydrolyzed products of alpha- and beta-tetra(L-aspartic acid)s by PAA hydrolase-1 has showed that PAA hydrolase-1 is capable of hydrolyzing only the specific amide bonds between beta-aspartic acid units . The RP-HPLC analysis of the enzymatic hydrolysis of beta-oligo(L-aspartic acid)s (4 and 5 mers) by PAA hydrolase-1 has suggested that the enzymatic hydrolysis of beta-oligo(L-aspartic acid)s occurs via an endo-mode cleavage . In contrast, PAA hydrolase-2 hydrolyzed both alpha- and beta-oligo(L-aspartic acid)s via an exo-mode cleavage to yield L-aspartic acid as a final product . A kinetic study on the enzymatic hydrolysis of alpha-oligo(L-aspartic acid)s (3 to 7 mers) by PAA hydrolase-2 has indicated that Km values are almost independent of the number of monomer units in oligomers of 4 to 7 mers, while that Vmax values are markedly dependent on the chain length and show a maximum value at 5 mer. Nat Immunol, 2004 Nov, 5(11), 1134 - 42 Epub 2004 Oct 03. CD28 induces immunostimulatory signals in dendritic cells via CD80 and CD86; Orabona C et al.; Bidirectional signaling along the B7-CTLA-4 coreceptor pathway enables reciprocal conditioning of T cells and dendritic cells . Although T cells can instruct dendritic cells to manifest tolerogenic properties after CTLA-4 engagement of B7, such a B7-mediated signaling is not known to occur in response to CD28 . Here we show that mouse dendritic cells were induced by soluble CD28 to express interleukin 6 and interferon-gamma . Production of interleukin 6 required B7-1 (CD80), B7-2 (CD86) and p38 mitogen-activated protein kinase and prevented interferon-gamma-driven expression of immunosuppressive tryptophan catabolism . In vivo, an adjuvant activity of soluble CD28 was demonstrated as enhanced T cell-mediated immunity to tumor and self peptides and protection against microbial and tumor challenge . Thus, different ligands of B7 can signal dendritic cells to express functionally distinct effector responses. J Exp Med, 2004 Oct 4, 200(7), 895 - 904 A subset of liver NK T cells is activated during Leishmania donovani infection by CD1d-bound lipophosphoglycan; Amprey JL et al.; Natural killer (NK) T cells are activated by synthetic or self-glycolipids and implicated in innate host resistance to a range of viral, bacterial, and protozoan pathogens . Despite the immunogenicity of microbial lipoglycans and their promiscuous binding to CD1d, no pathogen-derived glycolipid antigen presented by this pathway has been identified to date . In the current work, we show increased susceptibility of NK T cell-deficient CD1d(-/-) mice to Leishmania donovani infection and Leishmania-induced CD1d-dependent activation of NK T cells in wild-type animals . The elicited response was Th1 polarized, occurred as early as 2 h after infection, and was independent from IL-12 . The Leishmania surface glycoconjugate lipophosphoglycan, as well as related glycoinositol phospholipids, bound with high affinity to CD1d and induced a CD1d-dependent IFNgamma response in naive intrahepatic lymphocytes . Together, these data identify Leishmania surface glycoconjugates as potential glycolipid antigens and suggest an important role for the CD1d-NK T cell immune axis in the early response to visceral Leishmania infection. Appl Environ Microbiol, 2004 Oct, 70(10), 6037 - 46 Composition and diversity of microbial communities recovered from surrogate minerals incubated in an acidic uranium-contaminated aquifer; Reardon CL et al.; Our understanding of subsurface microbiology is hindered by the inaccessibility of this environment, particularly when the hydrogeologic medium is contaminated with toxic substances . In this study, surrogate geological media contained in a porous receptacle were incubated in a well within the saturated zone of a pristine region of an aquifer to capture populations from the extant communities . After an 8-week incubation, the media were recovered, and the microbial community that developed on each medium was compared to the community recovered from groundwater and native sediments from the same region of the aquifer, using 16S DNA coding for rRNA (rDNA)-based terminal restriction fragment length polymorphism (T-RFLP) . The groundwater and sediment communities were highly distinct from one another, and the communities that developed on the various media were more similar to groundwater communities than to sediment communities . 16S rDNA clone libraries of communities that developed on particles of a specular hematite medium incubated in the same well as the media used for T-RFLP analysis were compared with those obtained from an acidic, uranium-contaminated region of the same aquifer . The hematite-associated community formed in the pristine area was highly diverse at the species level, with 25 distinct phylotypes identified, the majority of which (73%) were affiliated with the beta-Proteobacteria . Similarly, the hematite-associated community formed in the contaminated area was populated in large part by beta-Proteobacteria (62%); however, only 13 distinct phylotypes were apparent . The three numerically dominant clones from the hematite-associated community from the contaminated site were affiliated with metal- and radionuclide-tolerant or acidophilic taxa, consistent with the environmental conditions . Only two populations were common to both sites. Appl Environ Microbiol, 2004 Oct, 70(10), 5868 - 74 Bacterial community structure and diversity in a century-old manure-treated agroecosystem; Sun HY et al.; Changes in soil microbial community structure and diversity may reflect environmental impact . We examined 16S rRNA gene fingerprints of bacterial communities in six agroecosystems by PCR amplification and denaturing gradient gel electrophoresis (PCR-DGGE) separation . These soils were treated with manure for over a century or different fertilizers for over 70 years . Bacterial community structure and diversity were affected by soil management practices, as evidenced by changes in the PCR-DGGE banding patterns . Bacterial community structure in the manure-treated soil was more closely related to the structure in the untreated soil than that in soils treated with inorganic fertilizers . Lime treatment had little effect on bacterial community structure . Soils treated with P and N-P had bacterial community structures more closely related to each other than to those of soils given other treatments . Among the soils tested, a significantly higher number of bacterial ribotypes and a more even distribution of the bacterial community existed in the manure-treated soil . Of the 99 clones obtained from the soil treated with manure for over a century, two (both Pseudomonas spp.) exhibited 100% similarity to sequences in the GenBank database . Two of the clones were possible chimeras . Based on similarity matching, the remaining 97 clones formed six major clusters . Fifty-six out of 97 were assigned taxonomic units which grouped into five major taxa: alpha-, beta-, and gamma-Proteobacteria (36 clones), Acidobacteria (16 clones), Bacteroidetes (2 clones), Nitrospirae (1 clone), and Firmicutes (1 clone) . Forty-one clones remained unclassified . Results from this study suggested that bacterial community structure was closely related to agroecosystem management practices conducted for over 70 years. Appl Environ Microbiol, 2004 Oct, 70(10), 5787 - 93 Strain-specific differences in the grazing sensitivities of closely related ultramicrobacteria affiliated with the Polynucleobacter cluster; Boenigk J et al.; Ultramicrobacteria (cell volume < 0.1 microm(3)) are the numerically dominant organisms in the plankton of marine and freshwater habitats . Flagellates and other protists are assumed to be the most important predators of these ultramicrobacteria as well as of larger planktonic bacteria . However, due to controversial observations conducted previously, it is not clear as to whether fractions of the ultramicrobacteria are resistant to flagellate predation . Furthermore, it is not known if closely related bacteria vary significantly in their sensitivity to flagellate predation . We investigated the sensitivity of ultramicrobacteria affiliated with the cosmopolitan Polynucleobacter cluster to grazing by Spumella-like nanoflagellates . Laboratory grazing experiments with four closely related (> or =99.6% 16S rRNA gene sequence similarity) bacteria and three closely related (100% 18S rRNA gene sequence similarity) flagellates were performed . In comparison to larger bacteria, predation on the ultramicrobacterial Polynucleobacter strains was weak, and the growth of the predating flagellates was slow . Specific clearance rates ranged between 0.14 x 10(5) and 2.8 x 10(5) units of predator size h(-1) . Feeding rates strongly depended on the flagellate and bacterial strain (P < 0.001) . Grazing mortality rates of the three flagellate strains investigated varied for the same prey strain by up to almost fourfold . We conclude that (i) ultramicrobacteria affiliated with the Polynucleobacter cluster are not protected from grazing, (ii) strain-specific variations in grazing sensitivity even between closely related bacteria are high, and (iii) strain-specific differences in predator-prey interaction could be an important factor in the evolution and maintenance of microbial microdiversity. Appl Environ Microbiol, 2004 Oct, 70(10), 5744 - 9 Ferrihydrite-dependent growth of Sulfurospirillum deleyianum through electron transfer via sulfur cycling; Straub KL et al.; Observations in enrichment cultures of ferric iron-reducing bacteria indicated that ferrihydrite was reduced to ferrous iron minerals via sulfur cycling with sulfide as the reductant . Ferric iron reduction via sulfur cycling was investigated in more detail with Sulfurospirillum deleyianum, which can utilize sulfur or thiosulfate as an electron acceptor . In the presence of cysteine (0.5 or 2 mM) as the sole sulfur source, no (microbial) reduction of ferrihydrite or ferric citrate was observed, indicating that S . deleyianum is unable to use ferric iron as an immediate electron acceptor . However, with thiosulfate at a low concentration (0.05 mM), growth with ferrihydrite (6 mM) was possible and sulfur was cycled up to 60 times . Also, spatially distant ferrihydrite in agar cultures was reduced via diffusible sulfur species . Due to the low concentrations of thiosulfate, S . deleyianum produced only small amounts of sulfide . Obviously, sulfide delivered electrons to ferrihydrite with no or only little precipitation of black iron sulfides . Ferrous iron and oxidized sulfur species were produced instead, and the latter served again as the electron acceptor . These oxidized sulfur species have not yet been identified . However, sulfate and sulfite cannot be major products of ferrihydrite-dependent sulfide oxidation, since neither compound can serve as an electron acceptor for S . deleyianum . Instead, sulfur (elemental S or polysulfides) and/or thiosulfate as oxidized products could complete a sulfur cycle-mediated reduction of ferrihydrite. Biochim Biophys Acta, 2004 Oct 11, 1685(1-3), 38 - 47 Metazoan and microbial models of Niemann-Pick Type C disease; Higaki K et al.; Niemann-Pick Type C (NP-C) disease compellingly provides insight into lipid transport and the association of this process with severe neuronal dysfunction . The two genes that define this syndrome, NPC1 and NPC2, are conserved throughout much of eukaryotic evolution, to the extent that the yeast and mammalian NPC1 genes are functionally interchangeable . We present here an evolutionary perspective of the genes defective in NP-C disease . We will describe how conservation of sequences and their biological roles in a variety of microbial and metazoan model systems may act as roadmaps to understanding this syndrome in humans. Colloids Surf B Biointerfaces, 2004 Oct 10, 38(1-2), 55 - 65 An electrodynamics-based model for ion diffusion in microbial polysaccharides; Liu C et al.; An electrodynamics-based model was formulated for simulation of ion diffusion in microbial polysaccharides . The fixed charges and electrostatic double layers that may associate with microbial polysaccharides and their effects on ion diffusion were explicitly built into the model . The model extends a common multicomponent ion diffusion formulation that is based on irreversible thermodynamics under a zero ionic charge flux condition, which is only applicable to the regions without fixed charges and electrostatic double layers . An efficient numerical procedure was presented to solve the differential equations in the model . The model well described key features of experimental observations of ion diffusion in negatively charged microbial polysaccharides including accelerated diffusive transport of cations, exclusion of anions, and increased rate of cation transport with increasing negative charge density . The simulated diffusive fluxes of cations and anions were consistent with a cation exchange diffusion concept in negatively charged polysaccharides at the interface of plant roots and soils; and the developed model allows to mathematically study such diffusion phenomena . An illustrative example was also provided to simulate dynamic behavior of ionic current during ion diffusion within a charged bacterial cell wall polysaccharide and the effects of the ionic current on the compression or expansion of the bacterial electrostatic double layer at the interface of the cell wall and bulk solution. Biochem Biophys Res Commun, 2004 Nov 5, 324(1), 108 - 13 Hinnuliquinone, a C2-symmetric dimeric non-peptide fungal metabolite inhibitor of HIV-1 protease; Singh SB et al.; HIV-1 protease is one of several key enzymes required for the replication and maturation of HIV-1 virus . An almost two-decade research effort by academic and pharmaceutical institutions resulted in the successful commercialization of seven drugs that are potent inhibitors of HIV-1 protease activity and which, if used correctly, are highly effective in managing viral load . However, identification of clinical viral isolates that are resistant to these drugs indicates that this is a significant problem and that new classes of inhibitors are continually needed . Screening of microbial extracts followed by bioassay-guided isolation led to the discovery of a natural hinnuliquinone, a C(2)-symmetric bis-indolyl quinone natural product that inhibited the wild-type and a clinically resistant (A44) strain of HIV-1 protease with K(i) values of 0.97 and 1.25microM, respectively . Crystallographic analysis of the inhibitor-bound HIV-1 protease helped explain the importance of the C(2)-symmetry of hinnuliquinone for activity . Details of the isolation, biological activity, and crystallographic analysis of the inhibitor-bound protease are herein described. Toxicology, 2004 Dec 15, 205(3), 201 - 10 Enhancement of the biodegradability of aromatic groundwater contaminants; Bittkau A et al.; Groundwater (GW) from the Bitterfeld industrial region, Central Germany, is contaminated mainly with monochlorobenzene (MCB) . Accordingly, current research addresses the development of feasible in situ groundwater remediation technologies . Although easily degradable under aerobic conditions, MCB persists in the essentially anaerobic Bitterfeld aquifer . Therefore, we focused on primary oxidation of MCB and the subsequent anaerobic biodegradability of MCB oxidation products by the indigenous microbial community . In groundwater microcosms, most efficient MCB removal was observed upon treatment with Fenton's reagent (H2O2 + Fe2+), which produces the highly reactive hydroxyl radical and Fe3+ simultaneously . Phospholipid fatty acid analysis following different treatments suggested respective shifts of the microbial community compositions, and indicated that Fenton's reagent had a rather beneficial than an adverse effect on biomass development . Potential metabolites of hydroxyl radical attack on MCB such as chlorohydroquinone, hydroquinone, catechol, resorcinol, and phenol were anaerobically degraded by the groundwater microbial community under Fe3+ -reducing conditions. J Biotechnol, 2004 Oct 19, 114(1-2), 1 - 9 Cloning and overexpression of the old yellow enzyme gene of Candida macedoniensis, and its application to the production of a chiral compound; Kataoka M et al.; The gene encoding old yellow enzyme (OYE), which catalyzes the conversion of ketoisophorone (KIP; 2,6,6-trimethyl-2-cyclohexen-1,4-dione) to (6R)-levodione (2,2,6-trimethylcyclohexane-1,4-dione), of Candida macedoniensis was cloned and sequenced . A 1212bp nucleotide fragment (oye) was confirmed to be the gene encoding OYE based on the agreement of internal amino acid sequences . Oye encodes a total 403 amino acid residues, and the deduced amino acid sequence shows a high degree of similarity to those of other microbial OYE family proteins . An expression vector, pETOYE, that contains the full length of oye was constructed . Escherichia coli harboring pETOYE exhibited an about six-fold increase in specific KIP-reducing activity under the control of the T7 promoter as compared with that of C . macedoniensis . (6R)-Levodione formed with washed cells of the transformant and a cofactor regeneration system amounted to 638 mM (98.2 mg ml(-1)), the a molar yield being 96.9% . The asymmetric reduction of KIP to (6R)-levodione with E . coli cells, which co-expressed both oye and the glucose dehydrogenase gene (gdh), as a catalyst was investigated . The (6R)-levodione formed amounted to 627 mM (96.6 mg ml(-1)), the a molar yield being 95.4% . Since the use of E . coli BL21 (DE3) cells co-expressing oye and gdh as a catalyst is simple and does not require the addition of glucose dehydrogenase, it is highly advantageous for the practical synthesis of (6R)-levodione. Dent Clin North Am, 2004 Oct, 48(4), 1077 - 104, ix Dental laser research: selective ablation of caries, calculus, and microbial plaque: from the idea to the first in vivo investigation; Rechmann P; One of the current dental laser research tracks is selective ablation, which is the removal of disease while not harming adjacent healthy structures . This article describes the scientific path from the first basic laboratory study of the absorption characteristics of caries to selective ablation of bacteria, microbial plaque, and calculus with a blue laser and the first in vivo safety studies in dogs . This article is an example of a typical research path for future studies of the uses of lasers in dentistry. Parasitol Today, 1993 Jun, 9(6), 201 - 6 The flagellar pocket of trypanosomatids; Webster P et al.; The surface of the trypanosomatid forms the interface between the parasite and its host, and has evolved to repel a variety of host anti-microbial defences . The flagellar pocket constitutes a highly differentiated region of the trypanosomatid surface that facilitates internalization of host macromolecules, while restricting host access to the exposed, endocytic receptors of the parasite . In this review, Paul Webster and David Russell discuss the ability of this organelle to accumulate efficiently nutrients obtained from the host as a major factor in the success of this group of parasites. J Liposome Res, 2004, 14(1-2), 77 - 86 Encapsulation of enrofloxacin in liposomes I: preparation and in vitro characterization of LUV; Sezer AD et al.; Liposomes are effectively used in the treatment of microbial infections . Higher cellular uptake has been reported when antibiotics are encapsulated in liposomes . In this study, enrofloxacin (ENF) was encapsulated in large unilamellar vesicles (LUVs) and the effects of formulation variables on the liposome characteristics were investigated . Liposomes were prepared using dry lipid film method . A number of variables such as molar ratios of phospholipid (DPPC; DL-alpha-phosphatidylcholine dipalmitoyl), cholesterol, ENF and amount of alpha-tocopherol and the volumes of internal (chloroform) and external phases {phosphate buffered saline PBS (pH 7.4)} were studied . In vitro characterization of the liposomes including the encapsulation capacity, size and drug release properties were carried out . Using of this method, spherical LUV liposomes with high drug content could be produced . Particle size of liposomes changed between 3.12 and 4.95 microm . The molar ratios of DPPC, cholesterol and ENF affected the size of the liposome (p < 0.05) . The drug encapsulation capacities were high and changed between 37.1% and 79.5% . The highest ENF encapsulation was obtained with the highest cholesterol content . An increase in the drug encapsulation capacity of the liposome was found with increasing molar ratios of DPPC, cholesterol and ENF (p < 0.05) . Furthermore, the release of ENF from the liposomes decreased as the molar ratios of DPPC, cholesterol and ENF increased (p < 0.05) . In conclusion, a convenient colloidal carrier for the controlled release of ENF can be prepared by changing the formulation parameters of LUVs. Int J Immunopathol Pharmacol, 2004 Sep-Dec, 17(3), 283 - 92 Augmentation of macrophage phagocytosis by modified arabinoxylan rice bran (MGN-3/biobran); Ghoneum M et al.; MGN-3/Biobran, modified arabinoxylan rice bran, has been shown to be a potent biological response modifier (BRM) as manifested by stimulation of different arms of the immune system such as NK, T and B cel