bluuebunny/crossref_metadata_2025 huggingface dataset:
Parotid secretory protein is expressed and inducible in human gingival keratinocytes
Authors: Shiba Hideki , Venkatesh S. G. , Gorr Sven‐Ulrik , Barbieri Germán , Kurihara Hidemi , Kinane Denis F.
Published: February 2005

Background:  Parotid secretory protein (PSP) is a major salivary protein that is thought to possess both antibacterial and anti‐inflammatory activity. A major question is whether PSP expression can be regulated by humoral factors and bacteria. Periodontitis is an inflammatory lesion initiated by interaction between gingival keratinocytes and periodontopathogenic microorganisms such as the Gram‐negative anaerobe Porphyromonas gingivalis. Cytokines and sex hormones have been implicated in the progression of various forms of periodontal diseases.

Materials and methods:  We investigated the expression of PSP and its regulation in primary cultures of human gingival keratinocytes (HGK). HGK at the third or fourth passage were exposed to heat‐killed P. gingivalis, tumor necrosis factor‐α (TNF‐α) and 17β‐estradiol. The PSP mRNA levels were examined by real‐time polymerase chain reaction (PCR). The protein expression of PSP was confirmed by immunofluorescence.

Results:  Heat‐killed P. gingivalis, TNF‐α and 17β‐estradiol all resulted in increased HGK levels of mRNA for PSP as determined by real‐time PCR analysis. Immunofluorescence demonstrated increased PSP localized within the cytoplasm of HGK following exposure to killed P. gingivalis.

Conclusion:  The present study has demonstrated for the first time that PSP is expressed in keratinocytes and that it can be up‐regulated by bacteria and humoral factors. Thus PSP may have a role in the innate defense system at the gingival epithelial surface.
Effectiveness of scaling and root planing versus modified Widman flap on nitric oxide synthase and arginase activity in patients with chronic periodontitis
Authors: Güllü Cigdem , Ozmeric Nurdan , Tokman Benay , Elgün Serenay , Balos Köksal
Published: February 2005

Background:  Nitric oxide (NO) is synthesized from the conversion of l‐arginine to l‐citrulline by NO synthase (NOS). Arginase, which is an arginine‐depleting enzyme, can compete with NOS for the common substrate l‐arginine and thus inhibit NO production.

Objectives:  In the present study, we aimed to examine the correlation between the arginase and NOS activity in patients with chronic periodontitis and to compare the effects of scaling and root planing and modified Widman flap procedures on enzyme activity.

Material and methods:  The study included 13 patients diagnosed with chronic periodontitis. Using a split‐mouth design, the defects showing ≥ 7 mm of attachment loss were treated either with scaling and root planing or with modified Widman flap. Gingival biopsies from both sites were obtained at baseline and 2 months after periodontal treatment. Immunohistochemical staining was performed for evaluating NOS expression and specific arginase activity was determined spectrophotometrically.

Results:  Although inflamed periodontal tissues demonstrated a strong inducible NOS (iNOS) expression at baseline, immunostaining decreased after periodontal treatment. iNOS expression intensity and the number of inflammatory cells showing iNOS expression were found to be higher in the scaling and root planing group compared to the modified Widman flap group. The specific activity of arginase was measured as 0.18 ± 0.07 IU/mg protein in the modified Widman flap group and 0.25 ± 0.11 IU/mg protein in the scaling and root planing group at baseline. After periodontal therapy, the enzyme level was increased to 0.68 ± 0.14 IU/mg protein in the modified Widman flap and to 1.10 ± 0.23 IU/mg protein in the scaling and root planing group.

Conclusion:  This study was the first report of evaluating the involvement of the arginine–NO pathway in chronic periodontitis and this might be considered to be of value in understanding the periodontal disease mechanisms.
Tumor Necrosis Factor‐α in the Placenta is not Elevated in Pre‐eclamptic Patients Despite its Elevation in Peripheral Blood
Authors: Hayashi Masatoshi , Ueda Yoshihiko , Yamaguchi Takehiko , Sohma Ryoichi , Shibazaki Mitsuei , Ohkura Takeyoshi , Inaba Noriyuki
Published: February 2005

Problem:  Tumor necrosis factor‐α (TNF‐α) is present in human placental and uterine cells at the early and late stages of gestation and promotes the regulation of trophoblast growth and invasion. We evaluated whether TNF‐α levels in the placenta and blood of pre‐eclamptic women differed from those with normal pregnancies.

Method of study:  The subjects were 39 pregnant women carrying single fetuses (21 normal‐pregnant and 18 pre‐eclamptic patients). Their average gestational age at entry was 38–39 weeks. Peripheral blood was collected before the onset of labor and separated serum was stored at −20°C. A tissue segment of the placenta was cut and frozen in liquid nitrogen immediately after delivery at −80°C. The frozen placental tissue was added to phosphate‐buffered saline. The tissue was fully homogenized and centrifuged. Separated supernatant was stored at −80°C. TNF‐α levels in separated serum and TNF‐α and total protein (TP) levels in separated supernatant were measured. The presence of TNF‐α in the placenta was evaluated by immunohistochemistry in five pre‐eclamptic and five normal‐pregnant patients.

Results:  Serum TNF‐α levels were higher in pre‐eclampsia than in normal pregnancies. However, TNF‐α/TP levels in the placenta did not differ significantly between the two groups. As for TNF‐α immunostaining of trophoblastic cells in the placenta, it was weak in three and moderate in two of the normal pregnancies, while it was absent in two, weak in one, and moderate in two in the pre‐eclampsia group.

Conclusions:  We demonstrated no significant increase in TNF‐α/TP levels in the placenta in pre‐eclampsia despite a significant increase in serum TNF‐α levels. There was no strong immunostaining for TNF‐α detected by immunohistochemistry in the pre‐eclampsia group. These findings suggest that TNF‐α in the placenta is not a key cytokine to interfere with normal trophoblast invasion into the myometrium in pre‐eclampsia, and that sources other than the placenta may contribute to the elevated levels of TNF‐α found in the circulation of pre‐eclamptic patients.
Increase of Activated Mast Cells in Human Endometriosis
Authors: Sugamata Masao , Ihara Tomomi , Uchiide Ichiro
Published: February 2005

Problem:  The proliferation of stromal cells in endometriosis promotes extensive adhesion; therefore, the morphological analysis of stromal lesions is important in the investigation of the pathogenesis of endometriosis.

Method of Study:  In this study, the morphological and numeric comparisons of degranulated mast cells were performed between endometriotic lesions and comparative regions (eutopic endometrium and normal uterine serosa) of patients with and without endometriosis.

Results:  In cases of endometriosis, diffuse infiltration of numerous mast cells was observed throughout the stromal lesions. These mast cells exhibited degranulation, and scattered granules were also observed. In the eutopic endometrium and normal uterine serosa of both the endometriosis patients and the controls, mast cells were rarely detected.

Conclusions:  These results suggest that an abnormal immune response, specifically a hypersensitivity reaction, is strongly related to endometriosis; our findings will be helpful in the development of methods for the treatment of endometriosis.
Preconception Peripheral Natural Killer Cell Activity as a Predictor of Pregnancy Outcome in Patients with Unexplained Infertility
Authors: Matsubayashi Hidehiko , Shida Masako , Kondo Akane , Suzuki Takahiro , Sugi Toshitaka , Izumi Shun‐Ichiro , Hosaka Takashi , Makino Tsunehisa
Published: February 2005

Problem:  Preconception high peripheral natural killer (NK) cell activity in women with recurrent spontaneous abortion can predict subsequent miscarriages. We have examined prospectively, for the first time, the pregnancy rate in patients with unexplained infertility by measuring the peripheral NK activity.

Method of study:  We tested the peripheral NK activity of 94 infertile women who despite treatment were unable to conceive for 6 or more months (mean; 2.4 years). Peripheral NK activity was measured by a chromium‐51 release cytotoxicity assay. Women were followed for 2 years and assessed.

Results:  In 77 patients who were followed for 2 years, 28 had conceived but 49 did not. The peripheral NK activity of the group that became pregnant (mean ± S.D.; 34.5 ± 13.8%) was significantly lower than that of non‐conception group (42.3 ± 13.3%, P = 0.017).

Conclusions:  Our finding suggests that elevated peripheral NK activity in patients with unexplained infertility is a risk factor for attaining pregnancy success.
Antiproliferative Actions of Ovine Uterine Serpin
Authors: Tekin Şaban , Padua Maria B. , Brad Amber M. , Hansen Peter J.
Published: February 2005

Problem:  Ovine uterine serpin (OvUS) is a member of the serine proteinase inhibitor superfamily and is the major protein produced by luminal and glandular epithelium of the sheep endometrium during mid to late pregnancy. The protein does not have prototypical proteinase inhibitory activity but can inhibit a wide variety of lymphocyte functions such as mitogen‐induced proliferation and natural killer cell cytotoxicity.

Method of study:  The antiproliferative actions of OvUS were studied.

Results:  It was demonstrated that, in addition to inhibiting lymphocyte proliferation, OvUS inhibits growth of two tumor cell lines (D17 and PC‐3). The protein also interrupts development of pre‐implantation embryos. Inhibition of cell proliferation is not universal, however, as OvUS did not inhibit growth of two non‐tumorigenic cell lines (MDBK and BEND). The mechanism of action of inhibitory effects of OvUS is not known although experiments with inhibitors of protein kinase A indicate that the protein does not inhibit lymphocyte proliferation through this pathway. Moreover, the protein does not induce apoptosis.

Conclusions:  The finding that OvUS has antiproliferative activity is demonstrative of the wide range of functions exerted by members of the serpin superfamily. The antiproliferative property of OvUS may reflect the role of the protein during pregnancy and may be exploitable for design of new antiproliferative drugs.
QUALITY, SENSORY AND COST COMPARISON FOR pH REDUCTION OF SYRAH WINE USING ION EXCHANGE OR TARTARIC ACID1
Authors: WALKER TERESA , MORRIS JUSTIN , THRELFALL RENEE , MAIN GARY
Published: February 2005

The pH reduction treatments (nonadjusted control, two resins in column systems, three membranes in a prototype electrochemical ion‐exchange unit, and tartaric acid addition) were evaluated on Syrah (Vitis vinifera) wine. Wine was analyzed after 6 months at 21C. Treatments lowered wine pH from 4.24 to 3.48, improved color attributes and raised titratable acidity, but lowered total phenolics and potassium levels. A descriptive sensory panel identified and evaluated purple color, aroma (berry, grape kool‐aid, dried fruit, spicy, barnyard/leather and smoky) and flavor (dried fruit, earthy, sourness/acidity and astringency) attributes of Syrah wine. The panel noticed more differences between the nonadjusted control and the pH reduction treatments rather than within the pH reduction treatments. A costing model of the treatments showed that tartaric acid addition for adjusting pH was more economical than ion‐exchange systems. Tartaric acid adjustment did not enhance the wine's color attributes as well as the ion‐exchange treatments. Although use of ion‐exchange systems are more costly than tartaric acid addition, ion exchange could be used to maintain or improve premium wine quality.
SENSORY PROFILING AND POSITIONING OF JILEBI SAMPLES BY MULTIVARIATE ANALYSIS
Authors: PRAKASH MAYA , RAVI RAMASAMY , DATTATREYA ANUPAMA , BHAT KODANGALA KESHAVA
Published: February 2005

Sensory profilograms based on the quantitative descriptive analysis (QDA) of jilebi using 12 attributes were drawn for nine market samples. The instrumental texture measurement of shear values and CIE color measurement for L*, a* and b* were determined for all the samples. Further, canonical discriminant analysis was run to position the samples. The results indicated that samples 9, 4 and 5 were grouped together in the same quadrant, which represented desirable attributes of jilebi viz, crispness, sweetness, glossy appearance and overall quality (OQ). Samples 6, 8 and 2 have taken a different quadrant with attributes like juicy and porous; while sample 3 was away in a separate quadrant with undesirable attribute namely heated oil. Crispness of jilebi had high positive correlation with OQ and texture (shear value).
Reversible Leukoencephalopathy Syndrome Associated to Leukopenia in a Chronic Hemodialysis Patient
Authors: Salgado Octavio J , Rosales Beatriz C , García Rafael
Published: February 2005

Abstract:  Reversible leukoencephalopathy syndrome (RLS) is a rare brain disorder, characterized by diffuse attenuation of cerebral white matter, which has been most commonly observed in transplant patients receiving calcineurin inhibitors or in patients with severe hypertension. We report an episode of RLS in a 22‐year‐old male patient on chronic hemodialysis with well‐controlled moderate hypertension who presented with de novo headache and generalized seizures. Cranial magnetic resonance image (MRI) revealed multiple areas of increased signal intensity in the white matter on T2‐weighed images which resolved spontaneously at subsequent MRIs. White blood cell count showed leucopenia with normal CD4 count at flow cytometry. A viral etiology could not be demonstrated. Reversible leukoencephaolopathy syndrome symptoms remitted within 72 h but leukopenia persisted over 10 months. The patient received a kidney transplant 15 months after RLS onset and has received cyclosporine since the second post‐transplant day. No recurrence of RLS symptoms has been observed. The etiology of the MRI changes in the present case seemed not to be either vasogenic or cytotoxic.
Problems in Adapting the K/DOQI Guidelines for Japanese Patients with End Stage Renal Failure
Authors: Yokoyama Keitaro
Published: February 2005

Abstract:  The National Kidney Foundation published The K/DOQI Clinical Practice Guidelines for Bone Metabolism and Disease in Chronic Kidney Disease. We should also consider differences in clinical features of management for renal osteodystrophy between Japan and the USA. The first problem is related to a difference in the method of Ca correction. The second problem is related to a difference in the timing of measurement. In the USA the timing of measurement varies with each institution. Briefly, it is performed at midweek (2 days after the last dialysis) in some of institutions, while at the beginning of next week (3 days after the last dialysis) in some others.

In addition, differences in the dose and type of the calcium‐containing phosphorus adsorbent and active vitamin D used should also be taken into consideration.
Surgical Indications and Procedures of Parathyroidectomy in Patients with Chronic Kidney Disease
Authors: Tominaga Yoshihiro , Matsuoka Susumu , Sato Tetsuhiko
Published: February 2005

Abstract:  The Kidney Foundation of the USA proposed clinical practice guidelines for bone metabolism and disease in chronic kidney disease including parathyroidectomy (PTx). We performed PTx in a total of 1725 patients with advanced secondary hyperparathyroidism (2HPT) and evaluated the K/DOQI guideline concerning PTx, comparing it with our surgical strategy. The guidelines emphasize the avoidance of ectopic calcification and cardiovascular complications which may be induced by hypercalcemia, hyperphosphatemia, and persistent high parathyroid hormone level. We agree with the attitude of the K/DOQI guideline. To decide surgical indications, we emphasize that the size of parathyroid gland is one of the important factors. The guideline recommends subtotal PTx and total PTx with autotransplantation. We prefer total PTx with forearm autograft, mainly because it is easier and safer to remove the residual parathyroid tissue from the forearm at recurrence compared to neck re‐exploration. In Japan, almost all patients require long‐term hemodialysis after PTx because of the very small opportunity of kidney transplantation. The risk of recurrence is not negligible. Based on our huge experience we believe our strategy for advanced secondary hyperparathyroidism can be accepted at least in Japan.
Effect of anti‐inflammatory and antioxidant drugs on the long‐term repair of severely injured mouse skeletal muscle
Authors: Vignaud A. , Cebrian J. , Martelly I. , Caruelle J.‐P. , Ferry A.
Published: February 2005

Non‐steroidal anti‐inflammatory drugs are frequently prescribed after skeletal muscle injury. It is not known whether this type of medication can interfere with muscle repair, although inflammatory response is thought to play an important role in this process. Tibialis anterior muscles of mice were injured by myotoxic agent (snake venom) or crushed. Then, animals were treated daily for 10–14 days with different types of non‐steroidal anti‐inflammatory and antioxidant drugs. The long‐term repair was studied 10–42 days after injury by analysing the recovery of in situ muscle force production, size of regenerating muscle cells and expression of myosin heavy chain. Our results show that diclofenac, diferuloylmethane (curcumin), dimethylthiourea or pyrrolidine dithiocarbamate treatment did not significantly affect muscle recovery after myotoxic injury (P > 0.05). Similarly, diferuloylmethane, dimethyl sulphoxide or indomethacin administration did not markedly change muscle repair after crush injury. However, we noted that high doses (> 2 mg kg−1) of diferuloylmethane or indomethacin increased lethality and reduced muscle repair after crush injury. In conclusion, non‐steroidal anti‐inflammatory and antioxidant drugs did not exhibit long‐term detrimental effects on muscle recovery after injury, except at lethal doses.
Influence of Transcriptional Activator RamA on Expression of Multidrug Efflux Pump AcrAB and Tigecycline Susceptibility in
Klebsiella pneumoniae
Authors: Ruzin Alexey , Visalli Melissa A. , Keeney David , Bradford Patricia A.
Published: February 2005

Tigecycline is an expanded broad-spectrum antibacterial agent that is active against many clinically relevant species of bacterial pathogens, including
Klebsiella pneumoniae
. The majority of
K. pneumoniae
isolates are fully susceptible to tigecycline; however, a few strains that have decreased susceptibility have been isolated. One isolate, G340 (for which the tigecycline MIC is 4 μg/ml and which displays a multidrug resistance [MDR] phenotype), was selected for analysis of the mechanism for this decreased susceptibility by use of transposon mutagenesis with IS
903
φ
kan
. A tigecycline-susceptible mutant of G340, GC7535, was obtained (tigecycline MIC, 0.25 μg/ml). Analysis of the transposon insertion mapped it to
ramA
, a gene that was previously identified to be involved in MDR in
K. pneumoniae.
For GC7535, the disruption of
ramA
led to a 16-fold decrease in the MIC of tigecycline and also a suppression of MDR.
Trans
-complementation with plasmid-borne
ramA
restored the original parental phenotype of decreased susceptibility to tigecycline. Northern blot analysis revealed a constitutive overexpression of
ramA
that correlated with an increased expression of the AcrAB transporter in G340 compared to that in tigecycline-susceptible strains. Laboratory mutants of
K. pneumoniae
with decreased susceptibility to tigecycline could be selected at a frequency of approximately 4 × 10
−8
. These results suggest that
ramA
is associated with decreased tigecycline susceptibility in
K. pneumoniae
due to its role in the expression of the AcrAB multidrug efflux pump.
Differential Bidirectional Transfer of Indinavir in the Isolated Perfused Human Placenta
Authors: Sudhakaran Sreeja , Ghabrial Hany , Nation Roger L. , Kong David C. M. , Gude Neil M. , Angus Peter W. , Rayner Craig R.
Published: February 2005

The protease inhibitor (PI) indinavir may be used in the management of human immunodeficiency virus (HIV) infection during pregnancy. Poor maternal-to-fetal transfer of indinavir has been reported previously, but the mechanisms of transfer remain unknown. The bidirectional transfer of indinavir was assessed in dually perfused, isolated human placentae. Term placentae (
n
= 5) were obtained from non-HIV-infected pregnant women. To investigate transport mechanisms, the steady-state transfer of indinavir was compared to those of antipyrine (a marker of passive diffusion) and [
3
H]vinblastine (a marker of P-glycoprotein [P-gp] transport) in the maternal-to-fetal and fetal-to-maternal directions in each placenta. Indinavir and antipyrine perfusate concentrations were determined by using reverse-phase, high-performance liquid chromatography; [
3
H]vinblastine concentrations were measured by liquid scintillation. The antipyrine transfer clearance in each direction did not differ (
P
= 0.76), a finding consistent with passive diffusion. However, the maternal-to-fetal transfer clearance of vinblastine, normalized to that of antipyrine (clearance index) (0.31 ± 0.05), was significantly lower than the fetal-to-maternal clearance index of vinblastine (0.67 ± 0.17;
P
= 0.017), suggesting the involvement of placental P-gp. Similarly, the maternal-to-fetal clearance index of indinavir (0.39 ± 0.09) was significantly lower than its fetal-to-maternal clearance index (0.97 ± 0.12;
P
< 0.001). These results represent the first evidence for differential transfer of a xenobiotic in the intact human placenta. The use of transport modulators to increase the maternal-to-fetal transfer of PIs as a possible strategy to reduce mother-to-child transmission of HIV warrants investigation.
In Vitro Activity and Mechanism of Action of Methylenecyclopropane Analogs of Nucleosides against Herpesvirus Replication
Authors: Kern Earl R. , Kushner Nicole L. , Hartline Caroll B. , Williams-Aziz Stephanie L. , Harden Emma A. , Zhou Shaoman , Zemlicka Jiri , Prichard Mark N.
Published: February 2005

We have reported previously that methylenecyclopropane analogs of nucleosides have excellent activity against certain members of the herpesvirus family. A second generation, the 2,2-bis-hydroxymethyl derivatives, were synthesized, and 18 compounds were tested for activity in vitro against herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), human and murine cytomegalovirus (HCMV and MCMV), varicella-zoster virus (VZV), and Epstein-Barr virus (EBV). Selected analogs were also evaluated against human herpesvirus type 6 (HHV-6) and HHV-8. None of the 18 compounds had activity against HSV-1 or HSV-2, but four were active against VZV by plaque reduction (PR) assay at 50% effective concentration (EC
50
) levels of ≤50 μM. Six of the 18 compounds were active against HCMV by cytopathic effect or PR assays with EC
50
s of 0.5 to 44 μM, and all were active against MCMV by PR (0.3 to 54 μM). Four of the compounds were active against EBV by enzyme-linked immunosorbent assay (<0.3 to 4.4 μM). Four compounds with CMV activity were also active against HHV-6A and HHV-6B (0.7 to 28 μM), and three compounds were active against HHV-8 (5.5 to 16 μM). One of these, ZSM-I-62, had particularly good activity against CMV, HHV-6, and HHV-8, with EC
50
s of 0.7 to 8 μM. Toxicity was evaluated in adherent and nonadherent cells, and minimal cytotoxicity was observed. Mechanism of action studies with HCMV suggested that these compounds are phosphorylated by the ppUL97 phosphotransferase and are potent inhibitors of viral DNA synthesis. These results indicate that at least one of these compounds may have potential for use in treating CMV and other herpesvirus infections in humans.
Susceptibilities of Several Clinical Varicella-Zoster Virus (VZV) Isolates and Drug-Resistant VZV Strains to Bicyclic Furano Pyrimidine Nucleosides
Authors: Andrei Graciela , Sienaert Rebecca , McGuigan Chris , Clercq Erik De , Balzarini Jan , Snoeck Robert
Published: February 2005

Varicella-zoster virus (VZV) is responsible for primary infections as well as reactivations after latency in the dorsal root ganglia. The treatment of such infections is mandatory for immunocompromised patients and highly recommended for elderly patients with herpes zoster infections (also called zona or shingles). The treatment of choice is presently based on four molecules, acyclovir (ACV), valaciclovir, famciclovir, and (in Europe) brivudine (BVDU). We present here our data on the antiviral activity of a new class of potent and selective anti-VZV compounds, bicylic pyrimidine nucleoside analogues (BCNAs), against a broad variety of clinical isolates and different drug-resistant virus strains. The results show that the BCNAs are far more potent inhibitors than ACV and BVDU against clinical VZV isolates as well as the VZV reference strains Oka and YS. The BCNAs were not active against ACV- and BVDU-resistant VZV strains bearing mutations in the viral thymidine kinase gene but kept their inhibitory potential against virus strains with mutations in the VZV DNA polymerase gene. Mutant virus strains selected in the presence of the BCNAs were solely cross-resistant to drugs, such as ACV and BVDU, that depend for their antiviral action on metabolic activation by the viral thymidine kinase.
High Prevalence of Markers for Sulfadoxine and Pyrimethamine Resistance in
Plasmodium falciparum
in the Absence of Drug Pressure in the Ashanti Region of Ghana
Authors: Marks Florian , Evans Jennifer , Meyer Christian G. , Browne Edmund N. , Flessner Christa , von Kalckreuth Vera , Eggelte Teunis A. , Horstmann Rolf D. , May Jürgen
Published: February 2005

Markers of
Plasmodium falciparum
resistance to chloroquine (CQ) and pyrimethamine-sulfadoxine (PYR-SDX) are widespread in areas where malaria is endemic. In an area where the use PYR-SDX is negligible, the Ashanti Region of Ghana, West Africa, adult individuals were enrolled in an analysis of CQ- and PYR-SDX-associated molecular resistance markers in 2001 (
n
= 177) and 2003 (
n
= 180). Parasite prevalence, as assessed by PCR assays, were 56.5 and 48.8% in 2001 and 2003, respectively. A high frequency of CQ, PYR, and SDX resistance markers was observed, whereby, as a weak trend, the frequency was higher in 2003. The quintuple combination of three
pfdhfr
mutations and two
pfdhps
mutations has previously been recognized to be the most important determinant of PYR-SDX resistance. Approximately 60% of parasite carriers harbored fourfold mutated parasites, indicative of a considerable risk for a switch to high-level PYR-SDX resistance in an area where the rate of PYR-SDX use is low. Among the factors contributing to the high frequency of PYR-SDX resistance-associated mutations are background use of PYR-SDX, past use of PYR for malaria prophylaxis, cross-resistance of trimethoprim with PYR, and the sufficient biological fitness of resistant parasites in the absence of drug pressure.
In Vitro Activity of Ramoplanin against
Clostridium difficile
, Including Strains with Reduced Susceptibility to Vancomycin or with Resistance to Metronidazole
Authors: Peláez T. , Alcalá L. , Alonso R. , Martín-López A. , García-Arias V. , Marín M. , Bouza E.
Published: February 2005

We evaluated the in vitro activity of ramoplanin, an antimicrobial compound that inhibits cell wall synthesis by acting at the level of lipid intermediate formation, against
Clostridium difficile.
We included strains with reduced susceptibilities to vancomycin (vancomycin-intermediate [Van
i
] strains) or with resistance to metronidazole (Mtz
r
), in order to assess the potential utility of ramoplanin for the treatment of
C. difficile
-associated diarrhea. We tested the activity of ramoplanin against a total of 105 nonduplicate clinical isolates of toxigenic
C. difficile
, including 8 Van
i
isolates and 6 Mtz
r
isolates, obtained from our laboratory. Ramoplanin was active against all strains tested at concentrations ranging from 0.03 to 0.5 μg/ml (MICs at which 50 and 90% of isolates were inhibited, 0.25 μg/ml; geometric mean MIC, 0.22 μg/ml). All isolates, independently of their levels of susceptibility to vancomycin or metronidazole, were considered susceptible to ramoplanin (MICs, ≤0.5 μg/ml).
Efficacy and Toxicity of Zinc Salts as Candidate Topical Microbicides against Vaginal Herpes Simplex Virus Type 2 Infection
Authors: Bourne Nigel , Stegall Rachael , Montano Raquel , Meador Michael , Stanberry Lawrence R. , Milligan Gregg N.
Published: February 2005

Zinc salt solutions administered as topical microbicides provided significant protection against herpes simplex virus type 2 infection in a mouse vaginal challenge model. However, at the therapeutic concentration, the salt solutions caused sloughing of sheets of vaginal epithelial cells. These observations limit the utility of zinc salts as microbicides and suggest that the application of zinc solutions to mucosal surfaces has the potential to cause damage that might increase susceptibility to secondary infections at a later time.
In Vitro Development of Resistance to Six and Four Fluoroquinolones in
Mycoplasma pneumoniae
and
Mycoplasma hominis
, Respectively
Authors: Gruson D. , Pereyre S. , Renaudin H. , Charron A. , Bébéar C. , Bébéar C. M.
Published: February 2005

Selection of resistant mutants in sequential subcultures with increasing concentrations of six and four different fluoroquinolones was studied for one reference strain each of
Mycoplasma pneumoniae
and
Mycoplasma hominis
, respectively. All fluoroquinolones tested selected for resistance, with alterations affecting the quinolone resistance-determining regions of the four target topoisomerase genes.
Antimicrobial Susceptibility of
Shigella sonnei
Isolates in Japan and Molecular Analysis of
S. sonnei
Isolates with Reduced Susceptibility to Fluoroquinolones
Authors: Hirose Kenji , Terajima Jun , Izumiya Hidemasa , Tamura Kazumichi , Arakawa Eiji , Takai Nobuko , Watanabe Haruo
Published: February 2005

We performed susceptibility testing with
Shigella sonnei
isolates from imported and domestic cases of infection in Japan during 2001 and 2002. Some
S. sonnei
isolates were resistant to nalidixic acid, tetracycline, and trimethoprim-sulfamethoxazole. Most of the nalidixic acid-resistant strains showed reduced susceptibility to fluoroquinolones but did not show fluoroquinolone resistance.
Efg1 Involved in Drug Resistance by Regulating the Expression of
ERG3
in
Candida albicans
Authors: Lo Hsiu-Jung , Wang Jang-Shiun , Lin Chih-Yang , Chen Chia-Geun , Hsiao Ting-Yin , Hsu Chia-Tung , Su Chia-Li , Fann Ming-Ji , Ching Yu-Tai , Yang Yun-Liang
Published: February 2005

The
ERG3
gene in
Candida
albicans
was identified as a gene whose mRNA level was higher in the
cph1
/
cph1 efg1
/
efg1
double mutant than in the wild-type cells. Further study showed that Efg1, but not Cph1, negatively regulated
ERG3
. Mutations in
EFG1
consistently increased the susceptibility of the cells to antifungal agents.
Potential Clindamycin Resistance in Clindamycin-Susceptible, Erythromycin-Resistant
Staphylococcus aureus
: Report of a Clinical Failure
Authors: Levin Todd P. , Suh Byungse , Axelrod Peter , Truant Allan L. , Fekete Thomas
Published: February 2005

The
erm
gene product confers clindamycin resistance on
Staphylococcus aureus
. We report a clindamycin clinical failure where resistance developed on therapy in a D-test-positive strain. D tests of 91 clindamycin-susceptible, erythromycin-resistant
S
.
aureus
isolates showed that 68% of methicillin-susceptible and 12.3% of methicillin-resistant
S
.
aureus
strains were D-test positive.
Disequilibrium in Distribution of Resistance Mutations among
Mycobacterium tuberculosis
Beijing and Non-Beijing Strains Isolated from Patients in Germany
Authors: Hillemann Doris , Kubica Tanja , Rüsch-Gerdes Sabine , Niemann Stefan
Published: February 2005

Genotypic analysis of 103 multidrug-resistant
Mycobacterium tuberculosis
strains isolated in Germany in 2001 revealed that mutations in codon 531 (75.7%) of the
rpoB
gene and codon 315 (88.4%) of the
katG
gene are most frequent. Beijing genotype strains (60.2% of all isolates) displayed a different distribution of resistance mutations than non-Beijing strains.
Mutational Analysis of Metronidazole Resistance in
Helicobacter pylori
Authors: Moore Jill M. , Salama Nina R.
Published: February 2005

Metronidazole is one of a few antibiotics effective in eliminating
Helicobacter pylori
infection of the human stomach. Several chromosomal loci have been implicated in resistance to this drug. Saturation transposon mutagenesis of the
H. pylori
genome revealed inactivation of the
rdxA
gene as uniquely able to confer metronidazole resistance.
Experimental Prediction of the Evolution of Ceftazidime Resistance in the CTX-M-2 Extended-Spectrum Beta-Lactamase
Authors: Welsh Kerry J. , Barlow Miriam , Tenover Fred C. , Biddle James W. , Rasheed J. Kamile , Clark Leigh Ann , McGowan John E.
Published: February 2005

We applied in vitro evolution to an
Escherichia coli
strain containing
bla
CTX-M-2
and obtained 10 independent mutant
bla
CTX-M-2
alleles that confer elevated resistance to ceftazidime (MIC ≥ 32 μg/ml) but lost the ability to confer resistance to cefepime. All alleles had a Pro-to-Ser substitution at position 167.
Susceptibility of
Streptococcus
mutans
and
Actinobacillus
actinomycetemcomitans
to Bactericidal Activity of Human β-Defensin 3 in Biological Fluids
Authors: Maisetta Giuseppantonio , Batoni Giovanna , Esin Semih , Raco Giorgio , Bottai Daria , Favilli Flavia , Florio Walter , Campa Mario
Published: February 2005

Bactericidal activity of human β-defensin 3 (hBD-3) against
Streptococcus
mutans
and
Actinobacillus
actinomycetemcomitans
was inhibited in a dose-dependent manner by the presence of saliva and/or serum. Increasing the concentration of hBD-3 partially overcame this inhibition. A fast bactericidal effect was observed against both bacterial strains, suggesting a potential therapeutic use for hBD-3 in the local treatment of oral infections.
In Vitro Activity of Tebipenem, a New Oral Carbapenem Antibiotic, against Penicillin-Nonsusceptible
Streptococcus pneumoniae
Authors: Kobayashi Reiko , Konomi Mami , Hasegawa Keiko , Morozumi Miyuki , Sunakawa Keisuke , Ubukata Kimiko
Published: February 2005

The in vitro activity of tebipenem (TBM), a new oral carbapenem antibiotic, against
Streptococcus pneumoniae
clinical isolates (
n
= 202) was compared with those of 15 reference agents. The isolates were classified into five genotypic classes after PCR identification of abnormal
pbp1a
,
pbp2x
, and
pbp2b
genes: (i) penicillin-susceptible
S. pneumoniae
(PSSP) isolates with no abnormal
pbp
genes (
n
= 34; 16.8%), (ii) genotypic penicillin-intermediate
S. pneumoniae
(gPISP) isolates with only an abnormal
pbp2x
gene [gPISP (
2x
)] (
n
= 48; 23.8%), (iii) gPISP isolates with abnormal
pbp1a
and
pbp2x
genes (
n
= 32; 15.8%), (iv) gPISP isolates with abnormal
pbp2x
and
pbp2b
genes (
n
= 16; 7.9%), and (v) genotypic penicillin-resistant
S. pneumoniae
(gPRSP) isolates with three abnormal
pbp
genes (
n
= 72; 35.6%). The majority of the strains tested had
mefA
(
n
= 59; 29.2%) or
ermB
(
n
= 91; 45%) gene-mediating macrolide resistance. For these isolates the MIC at which 90% of isolates are inhibited was significantly lower for TBM than for the reference oral antibiotics, as follows: 0.002 μg/ml for PSSP, 0.004 μg/ml for gPISP (
2x
), 0.016 μg/ml for gPISP (isolates with abnormal
pbp1a
and
pbp2x
genes and isolates with abnormal
pbp2x
and
pbp2b
genes), and 0.063 μg/ml for gPRSP. In addition, TBM showed excellent bactericidal activity against gPRSP isolates, which exhibited a 3-log
10
decrease within 2 h when they were incubated with a concentration greater than or equal to the MIC. Inhibition of cell wall synthesis toward the long axis and subsequent cell lysis were observed by scanning electron microscopy after a short-term exposure to TBM, unlike the effects seen with cephalosporins. These data suggest that TBM has potent activity against multidrug-resistant
S. pneumoniae
, the causative pathogen of community-acquired respiratory tract infections.
Comparative Bacteriological Efficacy of Pharmacokinetically Enhanced Amoxicillin-Clavulanate against
Streptococcus pneumoniae
with Elevated Amoxicillin MICs and
Haemophilus influenzae
Authors: Berry Valerie , Hoover Jennifer , Singley Christine , Woodnutt Gary
Published: February 2005

A new pharmacokinetically enhanced formulation of amoxicillin-clavulanate (2,000 mg of amoxicillin/125 mg of clavulanate twice a day; ratio 16:1) has been designed, with sustained-release technology, to allow coverage of bacterial strains with amoxicillin-clavulanic acid MICs of at least 4/2 μg/ml. The bacteriological efficacy of amoxicillin-clavulanate, 2,000/125 mg twice a day, ratio 16:1, was compared in a rat model of respiratory tract infection versus four other amoxicillin-clavulanate formulations: 8:1 three times a day (1,000/125 mg), 7:1 three times a day (875/125 mg), 7:1 twice a day (875/125 mg), and 4:1 three times a day (500/125 mg); levofloxacin (500 mg once a day); and azithromycin (1,000 mg on day 1 followed thereafter by 500 mg once a day). Bacterial strains included
Streptococcus pneumoniae
, with amoxicillin-clavulanic acid MICs of 2/1 (one strain), 4/2, or 8/4 μg/ml (three strains each), and
Haemophilus influenzae
, one β-lactamase-positive strain and one β-lactamase-negative, ampicillin-resistant strain. Animals were infected by intrabronchial instillation. Antibacterial treatment commenced 24 h postinfection, with doses delivered by computer-controlled intravenous infusion to approximate the concentrations achieved in human plasma following oral administration. Plasma concentrations in the rat corresponded closely with target human concentrations for all antimicrobials tested. Amoxicillin-clavulanate, 2,000/125 mg twice a day, ratio 16:1, was effective against all
S. pneumoniae
strains tested, including those with amoxicillin-clavulanic acid MICs of up to 8/4 μg/ml and against β-lactamase-producing and β-lactamase-negative ampicillin-resistant
H. influenzae
. These results demonstrate the bacteriological efficacy of pharmacokinetically enhanced amoxicillin-clavulanate 2,000/125 mg twice a day (ratio 16:1) against
S. pneumoniae
with amoxicillin-clavulanic acid MICs of at least 4/2 μg/ml and support clavulanate 125 mg twice a day as sufficient to protect against β-lactamase in this rat model.
In Vitro Pharmacodynamics of Amphotericin B, Itraconazole, and Voriconazole againstAspergillus,Fusarium, andScedosporiumspp
Authors: Lewis Russell E. , Wiederhold Nathan P. , Klepser Michael E.
Published: February 2005

We compared the in vitro pharmacodynamics of amphotericin B, itraconazole, and voriconazole againstAspergillus,Fusarium, andScedosporiumspecies with a combination of two non-culture-based techniques: the tetrazolium salt 2,3-bis-(2-methoxy-4-nitro-5-[(sulfenylamino)carbonyl]-2H-tetrazolium-hydroxide) (XTT) colorimetric reduction assay, and fluorescent microscopy with the cellular morbidity dye bis-(1,3-dibutylbarbituric acid) trimethine oxonol (DiBAC) to directly visualize hyphal damage. Amphotericin B exhibited species-specific concentration-dependent activity, with 50% effective concentrations (EC50s) ranging from 0.10 to 0.12 mg/ml forA. fumigatus, 0.36 to 0.53 mg/ml forA. terreus, 0.27 to ≥32 mg/ml forF. solani, 0.41 to 0.55 mg/ml forF. oxysporum, and 0.97 and 0.65 mg/ml forS. apiospermumandS. prolificans, respectively. Similarly, itraconazole inhibited the growth ofA. fumigatusandA. terreusisolates with MICs of <1 mg/ml (EC500.03 to 0.85 mg/ml) andS. apiospermum, but was not active againstFusariumspecies orS. prolificans. Voriconazole effectively inhibited the growth ofAspergillus,Fusarium, andS. apiospermum(EC500.10 to 3.3 mg/ml) but had minimal activity against a multidrug-resistant isolate ofF. solaniorS. prolificans. Hyphal damage visualized by DiBAC staining was observed more frequently with voriconazole and amphotericin B versus itraconazole. These data highlight the species-specific differences in antifungal pharmacodynamics between mold-active agents that could be relevant for the development of in vitro susceptibility breakpoints and antifungal dosing in vivo.
Phase I Evaluation of the Safety and Pharmacokinetics of Murine-Derived Anticryptococcal Antibody 18B7 in Subjects with Treated Cryptococcal Meningitis
Authors: Larsen Robert A. , Pappas Peter G. , Perfect John , Aberg Judith A. , Casadevall Arturo , Cloud Gretchen A. , James Robert , Filler Scott , Dismukes William E.
Published: February 2005

A promising approach to improving outcomes in patients with cryptococcal meningitis is to use adjunctive passive immunotherapy with a monoclonal antibody (MAb) directed against the capsular polysaccharide ofCryptococcus neoformans. This is the first application of MAb therapy for the treatment of a fungal disease in humans. We determined the safety and maximum tolerated dose of the murine anticryptococcal MAb 18B7 in a phase I dose-escalation study. The subjects were human immunodeficiency virus-infected patients who had been successfully treated for cryptococcal meningitis. Six dosing cohorts received MAb 18B7 at 0.01 to 2 mg/kg of body weight as a single infusion. Three patients each received 0.01, 0.05, 0.2, and 0.5 mg of MAb 18B7 per kg without significant adverse events. Four of the subjects who received the 1-mg/kg dose had mild study drug-associated toxicity, including transient nausea, vomiting, back pain, and urticarial rash. Two of the subjects who received 2 mg/kg developed drug-associated mild to moderate nausea, vomiting, chills, and myalgias. One of the subjects who received 2 mg/kg developed intracranial hypertension 10 weeks after MAb 18B7 administration. Serum cryptococcal antigen titers in the cohorts receiving doses of 1 and 2 mg/kg declined by a median of twofold at 1 week and a median of threefold at 2 weeks postinfusion, but the titers subsequently returned toward the baseline values by week 12. The half-life of MAb 18B7 in serum was approximately 53 h, while the MAb was undetectable in the cerebrospinal fluid of all patients. These data support the continued investigation of MAb 18B7 at a maximum single dose of 1.0 mg/kg.
Small-Molecule Inhibitors of the Budded-to-Hyphal-Form Transition in the Pathogenic Yeast
Candida albicans
Authors: Toenjes Kurt A. , Munsee Suzanne M. , Ibrahim Ashraf S. , Jeffrey Rachel , Edwards John E. , Johnson Douglas I.
Published: February 2005

The pathogenic yeast
Candida albicans
can exist in multiple morphological states, including budded, pseudohyphal, and true hyphal forms. The ability to convert between the budded and hyphal forms, termed the budded-to-hyphal-form transition, is important for virulence and is regulated by multiple environmental and cellular signals. To identify inhibitors of this morphological transition, a microplate-based morphological assay was developed. With this assay, the known actin-inhibiting drugs latrunculin-A and jasplakinolide were shown to inhibit the transition in a dose-dependent and reversible manner. Five novel small molecules that reversibly inhibited the transition and hyphal elongation without affecting budded growth were identified. These molecules inhibited hyphal growth induced by Spider, Lee's, M199 pH 8, and 10% serum-containing media, with two molecules having a synergistic effect. The molecules also differentially affected the hyphal form-specific gene expression of
HWP1
and endocytosis without disrupting the actin cytoskeleton or septin organization. Structural derivatives of one of the molecules were more effective inhibiters than the original molecule, while other derivatives had decreased efficacies. Several of the small molecules were able to reduce
C. albicans
-dependent damage to endothelial cells by inhibiting the budded-to-hyphal-form transition. These studies substantiated the effectiveness of the morphological assay and identified several novel molecules that, by virtue of their ability to inhibit the budded-to-hyphal-form transition, may be exploited as starting points for effective antifungal therapeutics in the future.
Individual variation in the erythropoietic response to altitude training in elite junior swimmers
Authors: Friedmann B , Frese F , Menold E , Kauper F , Jost J , Bärtsch P
Published: February 2005

Objectives:Inter-individual variations in sea level performance after altitude training have been attributed, at least in part, to an inter-individual variability in hypoxia induced erythropoiesis. The aim of the present study was to examine whether the variability in the increase in total haemoglobin mass after training at moderate altitude could be predicted by the erythropoietin response after 4 h exposure to normobaric hypoxia at an ambient Po2corresponding to the training altitude.

Methods:Erythropoietin levels were measured in 16 elite junior swimmers before and after 4 h exposure to normobaric hypoxia (Fio20.15, ∼2500 m) as well as repeatedly during 3 week altitude training (2100–2300 m). Before and after the altitude training, total haemoglobin mass (CO rebreathing) and performance in a stepwise increasing swimming test were determined.

Results:The erythropoietin increase (10–185%) after 4 h exposure to normobaric hypoxia showed considerable inter-individual variation and was significantly (p<0.001) correlated with the acute erythropoietin increase during altitude training but not with the change in total haemoglobin mass (significant increase of ∼6% on average). The change in sea level performance after altitude training was not related to the change in total haemoglobin mass.

Conclusions:The results of the present prospective study confirmed the wide inter-individual variability in erythropoietic response to altitude training in elite athletes. However, their erythropoietin response to acute altitude exposure might not identify those athletes who respond to altitude training with an increase in total haemoglobin mass.
Proprioceptive neuromuscular facilitation training induced alterations in muscle fibre type and cross sectional area
Authors: Kofotolis N , Vrabas I S , Vamvakoudis E , Papanikolaou A , Mandroukas K
Published: February 2005

Objectives:To compare the effects of proprioceptive neuromuscular facilitation (PNF) and isokinetic training on fibre type distribution and cross sectional area of the vastus lateralis muscle.

Methods:Twenty four male university students were divided into two equal groups: PNF training and isokinetic training (ISO). The training regimen for the PNF group consisted of three sets of 30 repetitions against maximal resistance, alternating two patterns of sequential movements of the right lower extremity: (a) toe flexion and ankle plantar flexion and eversion; (b) knee extension and hip extension, abduction, and internal rotation. The ISO group performed three sets of 30 repetitions alternating knee extension and flexion of the right leg at angular velocities of 180 and 90°/s in an isokinetic dynamometer (Cybex). Both groups trained three times a week for a total of eight weeks. Muscle biopsy specimens were obtained from the right vastus lateralis muscle before and after training.

Results:The mean percentage area of type IIB fibre was significantly decreased (p<0.01) after eight weeks of PNF training, whereas that of type IIA fibre was significantly (p<0.05) increased. The mean percentage area of ISO trained type IIAB fibres exhibited an augmentative pattern (p<0.01) with a parallel reduction (p<0.05) in type IIA. Percentage fibre type distribution exhibited a similar pattern.

Conclusions:Both PNF and ISO training alter fibre type distribution and mean cross sectional area. These changes occur in the type II fibre subgroup.