Identification of genes differentially expressed in T cells following stimulation with the chemokines CXCL12 and CXCL10.

BACKGROUND: Chemokines are involved in many biological activities ranging from leukocyte differentiation to neuronal morphogenesis. Despite numerous reports describing chemokine function, little is known about the molecular changes induced by cytokines. METHODS: We have isolated and identified by differential display analysis 182 differentially expressed cDNAs from CXCR3-transfected Jurkat T cells following treatment with CXCL12 or CXCL10. These chemokine-modulated genes were further verified using quantitative RT-PCR and Western blot analysis. RESULTS: One hundred and forty-six of the cDNAs were successfully cloned, sequenced, and identified by BLAST. Following removal of redundant and non-informative clones, seventeen mRNAs were found to be differentially expressed post treatment with either chemokine ligand with several representing known genes with established functions. Twenty-one genes were upregulated in these transfected Jurkat cells following both CXCL12 and CXCL10, four genes displayed a discordant response and seven genes were downregulated upon treatment with either chemokine. Identified genes include geminin (GEM), thioredoxin (TXN), DEAD/H box polypeptide 1 (DDX1), growth hormone inducible transmembrane protein (GHITM), and transcription elongation regulator 1 (TCERG1). Subsequent analysis of several of these genes using semi-quantitative PCR and western blot analysis confirmed their differential expression post ligand treatment. CONCLUSIONS: Together, these results provide insight into chemokine-induced gene activation and identify potentially novel functions for known genes in chemokine biology.

Blood pressure outcomes of dental patients screened chronobiologically: a seven-year follow-up.

BACKGROUND: This article is a follow-up on a 1998 article in JADA. The blood pressure, or BP, of 23 of 24 normotensive patients was monitored at 15-minute intervals for a total of nine days, in three consecutive sessions of four, two and three days, respectively, separated by a few weeks. METHODS: Twelve patients were reached by phone seven years after the prior chronobiologically interpreted monitoring to ascertain their cardiovascular status since the initial monitoring. RESULTS: Only two of the 12 patients reached in follow-up had abnormalities in all three sessions, and only these two patients reported having experienced an adverse vascular event (one a myocardial infarction, the other coronary artery bypass graft surgery). The difference in outcome between the patients with chronobiological abnormality in all three sessions vs. the pool of those with abnormality in only two, one or none of the sessions is statistically significant. CONCLUSIONS: Even a five-day (and sometimes a longer) profile, while greatly preferred to single measurements, may not suffice for a definitive diagnosis of certain patients. Retrospectively, the 864 measurements, on the average, on each person in our study could have served for a recommendation to each person. Chronobiologically interpreted BP and heart rate monitoring for a week or longer as a start detects high-risk states that may be missed by conventional casual measurements that are rarely accompanied by one-day profiles. CLINICAL IMPLICATIONS: The dentist has an important opportunity for conveying the importance of monitoring BP and heart rate for a week to detect an abnormal variability, and for implementing this monitoring. Dentists can educate patients regarding the need for screening and for interpreting the results for variability, and contribute to their overall health by referring them to their physicians when treatment is indicated.

Selenium-enriched Agaricus bisporus mushrooms suppress 7,12-dimethlybenz[a]anthracene bioactivation in mammary tissue.

The present studies compared dietary Se (1.0 microg/g) when provided as either fortified Agaricus bisporus mushrooms, or sodium selenite on the in vivo metabolism of 7,12-dimethylbenz(a)anthracene (DMBA). Dietary addition of Se unenriched A. bisporus mushrooms at 2% did not alter the occurrence of DMBA induced DNA adducts or the activity of glutathione S-transferase (GST). However, the addition of Se as enriched mushrooms, or as selenite, significantly increased both liver and mammary GST activity. Providing sodium selenite, or enriched mushrooms also significantly reduced total and anti-3,4-dihydrodiol-1,2-epoxide-deoxyguanosine adducts compared to feeding the basal diet (P < 0.05). These investigations provide evidence that Se enriched mushrooms can be used as an effective method to retard chemically induced tumors.

Interpreting vital sign profiles for maximizing patient safety during dental visits.

Concerns about local anesthetics and dental treatment stress are still prevalent in today's medical and dental communities. The authors conducted 24-hour monitoring of heart rate and blood pressure for 23 patients undergoing two root planing procedures and a single periodontal surgical procedure. Chronobiologic analysis of circadian blood pressure and heart rate rhythms revealed that neither epinephrine vasoconstrictor in anesthetics nor increased invasiveness of the procedure resulted in statistically significant deviations from regular, individualized blood pressure rhythms. For surgical appointments, time of day had the greatest effect on cardiovascular variables, with the greatest elevations in blood pressure seen for surgery at 8 a.m. and the largest blood pressure decreases seen for surgery at 2 p.m.

Garlic powder and allyl sulfur compounds enhance the ability of dietary selenite to inhibit 7,12-dimethylbenz[a]anthracene-induced mammary DNA adducts.

These studies examined the ability of garlic powder or allyl sulfur compounds to modify selenite protection against 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary epithelial cell DNA adducts. In Study 1, female rats (n = 5) were fed diets containing sodium selenite at 0.1, 0.5, or 1.0 mg Se/kg and garlic powder at 0, 20, or 40 g/kg diet. Total DNA adducts correlated inversely with selenite or garlic powder intake. Garlic powder enhanced the selenite inhibition of mammary DNA adducts. In Study 2, selenite (2.0 mg Se/kg diet), garlic powder (20 g/kg diet), water-soluble S-allyl cysteine (SAC; 5.2 mumol/kg diet), and oil-soluble diallyl disulfide (DADS; 5.2 mumol/kg diet) inhibited (p < 0.05) total DNA adducts by 45%, 40%, 80%, and 75%, respectively. Combining selenite with garlic powder, SAC, or DADS further inhibited DNA adducts. Selenite, but not garlic powder, SAC, or DADS, enhanced liver glutathione S-transferase and uridine diphosphate-glucuronosyltransferase activities. Selenite, garlic powder, SAC, or DADS did not affect liver cytochrome P-450 1A1 activities. The present studies provide evidence that synergistic protection against the initiation of DMBA carcinogenesis occurs when selenite is supplemented in conjunction with garlic or its allyl sulfur components.

Impact of dietary fatty acids on 7,12-dimethylbenz[a]anthracene-induced mammary DNA adducts.

These studies examined the impact of dietary corn oil and its major constituent fatty acids on the occurrence of 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary DNA adducts. In study 1, rats were fed diets containing 5, 10 or 20% corn oil for 2 weeks prior to DMBA treatment (25 mg/kg). Mammary DNA adducts increased significantly (P < 0.05) as the quantity of dietary corn oil increased. Liver glutathione S-transferase (GST) activities increased while UDP-glucuronosyltransferase (UGT) activities decreased as the quantity of dietary corn oil increased. Increased adducts resulting from greater corn oil consumption were positively correlated with GST and negatively correlated with UGT activities. In study 2, rats were fed diets containing 5 or 20% corn oil, or 5% corn oil supplemented with 1.67% palmitic, 3.81% oleic or 8.78% linoleic acid (quantities in 15% corn oil) for 2 weeks prior to DMBA treatment (50 mg/kg). Total mammary DNA adducts were 75, 136 and 156% greater in rats fed the 20% corn oil, oleic acid-supplemented and linoleic acid-supplemented diets, respectively, than in those fed the 5% corn oil diet. Palmitic acid supplementation did not affect the occurrence of adducts. Adducts in study 2 did not correlate with GST or UGT activities. These studies demonstrate that enhanced DMBA bioactivation caused by increased corn oil consumption can be at least partially explained by increased intake of oleic and linoleic acids.

Dietary components modify the ability of garlic to suppress 7,12-dimethylbenz(a)anthracene-induced mammary DNA adducts.

Various dietary components were evaluated as factors influencing garlic's ability to depress rat mammary cell DNA adducts resulting from 7,12-dimethylbenz(a)anthracene (DMBA) treatment. Diets with or without garlic powder (20 g/kg) were provided for 2 wk before DMBA treatment (25 mg/kg body weight). Rats fed diets containing 36 g casein/100 g diet had 31% fewer (P < 0.05) mammary cell DNA adducts than those fed 12 g/100 g. Garlic supplementation significantly (P < 0.05) reduced DNA adducts in rats fed either 12 or 36 g casein/100 g by 35 and 32% respectively. In the absence of dietary garlic, DNA adducts were 23% lower (P < 0.05) in rats provided a diet containing supplemental L-methionine at 0.9 g/100 g than at 0.3 g/100 g. However, adduct inhibition by garlic supplementation was greater in rats fed the lower (P < 0.05) amount of methionine (54 vs. 26% inhibition). Adduct levels in rats fed diets with 20 g corn oil/100 g were twice those occurring in rats fed 5 g/100 g (P < 0.05), regardless of adjustment for energy density. Garlic supplementation prevented the increase in DNA adducts caused by increasing dietary corn oil. Combining dietary supplements of garlic, selenite (0.5 mg/kg diet) and retinyl acetate (328 mg/kg diet) inhibited the occurrence of DNA adducts to a greater degree than when each was supplied individually. These studies demonstrate that while dietary garlic can reduce DNA adduct formation in mammary tissue caused by DMBA, this protection is influenced by several dietary components.

Garlic and associated allyl sulfur components inhibit N-methyl-N-nitrosourea induced rat mammary carcinogenesis.

Our previous studies demonstrated that dietary garlic powder supplementation inhibits N-nitrosamine induced DNA alkylation in liver and mammary tissue. The present studies compared the impact of dietary supplementation with garlic powder or two garlic constituents, water-soluble S-allyl cysteine (SAC) and oil-soluble diallyl disulfide (DADS), on the incidence of mammary tumorigenesis induced by N-methyl-N-nitrosourea (MNU). Female Sprague-Dawley rats were fed semi-purified casein based diets with or without supplements of garlic powder(20g/kg), SAC (57 micromol/kg) or DADS (57 micromol/kg) for 2 weeks prior to treatment with MNU (15 mg/kg body wt). Garlic powder, SAC and DADS supplementation significantly delayed the onset of mammary tumors compared to rats receiving the unsupplemented diet. Tumor incidence 23 weeks after MNU treatment was reduced by 76, 41 and 53% in rats fed garlic, SAC and DADS, respectively, compared to controls (P<0.05). Total tumor number was reduced 81, 35 and 65% by these supplements, respectively (P<0.05). In a separate study the quantity of mammary DNA alkylation occurring 3 h after MNU treatment was reduced in rats fed garlic, SAC or DADS (P<0.05). Specifically, O(6)-methylguanine adducts were reduced by 27, 18 and 23% in rats fed supplemental garlic, SAC and DADS, respectively, compared to controls. N(7)-Methylguanine adducts decreased by 48, 22 and 21% respectively, compared to rats fed the control diet. These studies demonstrate that garlic and associated allyl sulfur components, SAC and DADS, are effective inhibitors of MNU-induced mammary carcinogenesis.

The prevention of localized ridge deformities using guided tissue regeneration.

Healing of extraction sockets where the buccal plate of bone has been destroyed often results in localized ridge deformities. This is most likely due to the ingrowth of extraskeletal connective tissue into the space originally occupied by the tooth which then fills with a combination of bone and soft tissue. Techniques for managing these defects have been described, but it would be preferable to prevent them from occurring at all. Using the principles of guided tissue regeneration, a surgical technique is described that can be applied to post-extraction sites that are deficient in alveolar bone thereby preventing localized ridge deformities. Representative cases are presented illustrating the technique.

N-(4-hydroxyphenyl)retinamide (fenretinide) induces retinol-binding protein secretion from liver and accumulation in the kidneys in rats.

The chemopreventive retinoid N-(4-hydroxyphenyl)retinamide (HPR) depresses serum retinol and retinol-binding protein (RBP) concentrations. To study long-term effects of HPR on serum proteins, rats were fed a control diet or a diet containing HPR (737 mumol/kg diet) for 14 d. Serum retinol and RBP of HPR-treated rats decreased to 42 and 41%, respectively, of initial concentrations. Transthyretin, albumin and transferrin did not differ between treatments. Previous studies found that HPR decreased secretion of the retinol-RBP complex into plasma. To investigate acute effects of HPR on RBP metabolism, vitamin A-deficient rats were injected with HPR (51 mumol/kg body wt), retinol (0.52 mumol/rat) or Tween carrier only. Liver RBP concentrations in HPR- and retinol-treated rats were 45 and 18%, respectively, of concentrations in Tween-treated rats, indicating rapid RBP secretion. Tween- and HPR-treated rats maintained relatively constant serum RBP concentrations, whereas retinol-replete rats had 12-fold higher serum RBP after 150 min. Rats treated with HPR and rats treated with retinol had 29- and eightfold higher kidney RBP concentrations, respectively, than Tween-treated rats, indicating rapid clearance of RBP from plasma. We conclude that HPR affects RBP metabolism by inducing secretion of liver RBP into the bloodstream and rapid RBP accumulation in the kidney.

Improved pontic/tissue relationships using porous coralline hydroxyapatite block.

Porous coralline hydroxyapatite blocks were implanted in the anterior region of several patients to improve the pontic/ridge relationship for a fixed prosthesis, thereby improving esthetic appearance and oral hygiene performance. A beveled incision on the palate was used to displace the flap coronofacially without leaving an area of denuded bone or graft. Flap margins were sutured to the adjacent palatal tissues. At the 2 1/2- and 3-year postgrafting follow-up, the implant area was healthy and ridge contours have been maintained. The permanent fixed partial denture is functional, and radiographic evaluation indicates the continued presence of the porous coralline hydroxyapatite implant. Three-year results have been promising enough to warrant continuing the procedure in patients requiring localized ridge augmentation.

Porous hydroxyapatite grafts in chronic subcrestal periodontal defects in rhesus monkeys: a histological investigation.

The purpose of this investigation was to evaluate the histologic response to replamineform (porous) hydroxyapatite grafts in chronically inflamed, surgically created, periodontal pockets in Rhesus monkeys. Forty-eight subcrestal defects were surgically created in 8 Rhesus monkeys. Orthodontic wires were adapted to the base of the defects and left in place for 8 weeks. The defects were randomly assigned a treatment modality so that 8 granular porous and 8 block porous hydroxyapatite grafts were done. Eight sites were left as unoperated controls. Twenty-four contralateral sham-operated sites were treated by open curettage. Histometric measurements of the length of regenerated connective tissue attachment were recorded. Block hydroxyapatite, granular hydroxyapatite, and sham-operated sites exhibited similar amounts of regenerated connective tissue attachment, which was significantly greater than that which occurred in unoperated controls. Newly formed bone was observed within the porous channels of the hydroxyapatite grafts as well as in direct apposition on the surface of implant particles. New bone, cementum, and periodontal ligament was observed in grafted sites and in defects treated by open curettage. In some instances healing occurred by means of a long junctional epithelium. Histology confirmed that unoperated defects remained as unhealed, inflamed periodontal pockets. The results of this study suggest that porous hydroxyapatite grafts have the potential to regenerate the attachment apparatus in primates.

Four-year investigation of salt and peroxide regimen compared with conventional oral hygiene.

This study of 171 subjects compared the effectiveness of a patient-applied salt and peroxide oral hygiene regimen with the use of conventional oral hygiene methods on clinical measures of periodontal disease, and determined patient compliance with and acceptance of using these two oral hygiene regimens. Results indicated that both regimens, when combined with professional care, were effective in changing clinical measures of periodontal disease to a state favoring periodontal health.

Salt and peroxide compared with conventional oral hygiene. I. Clinical results.

The purposes of this 2-year longitudinal study were to: compare the clinical effectiveness of patient applied sodium bicarbonate, hydrogen peroxide, and sodium chloride (S/P) to the use of conventional oral hygiene methods and to investigate the motivational effect of using phase-contrast microscopy in teaching effective oral hygiene. Initially, 972 subjects were screened for signs of periodontitis. From these, 347 with early to moderate periodontitis were selected and each was randomly assigned to one of four home treatment regimens after scaling and root planing. The four treatment regimens included: conventional oral hygiene procedures, conventional oral hygiene procedures plus phase-contrast demonstration of subgingival microbial forms for oral hygiene motivation, S/P oral hygiene, and S/P oral hygiene plus phase-contrast demonstration of subgingival microbial forms for oral hygiene motivation. Plaque, bleeding, gingival inflammation, probing depth, and clinical attachment level were recorded at baseline, 8, 16, and 24 months. Subjects were recalled for reinforcement of oral hygiene and periodontal prophylaxis at various intervals. Data were analyzed based on disease severity, location of index sites and compliance. The results indicated that both conventional oral hygiene procedures and the S/P regimen were effective in reducing clinical signs of disease when combined with professional care. There were no differences between the two regimens in clinical effectiveness and trends favoring microscopic viewing of subgingival plaque for motivational purposes were not statistically significant.

Salt and peroxide compared with conventional oral hygiene. II. Microbial results.

This study was designed to investigate the effect of conventional oral hygiene (n = 116 subjects) versus a salt and peroxide oral hygiene regimen (n = 115 subjects) on subgingival microorganisms. Subgingival plaque for microscopic evaluation was obtained from eight index tooth sites in each of 231 adult subjects. Microbial forms were microscopically identified at baseline, 8, 16, and 24 months. For both oral hygiene groups, cocci were increased (P less than 0.05) and motile rods were decreased (P less than 0.05) at 8 months and returned to baseline by 16 months. Spirochetes were decreased (P less than 0.05) and remained low through 24 months in both oral hygiene groups. The frequency of agreement between clinical (bleeding) and microbial (greater than or equal to 15% spirochetes or motile rods or greater than or equal to 20% spirochetes + motile rods) criteria for instrumentation was 59.8%. It was also found that fewer total instrumentations for test subjects were observed when microbiological criteria were used as compared with clinical criteria. The greater number of instrumentations based on clinical criteria was highly significant (P less than or equal to 0.001). A significant change in microbial signs associated with peridontal disease may be obtained with either a conventional oral hygiene or a salt and peroxide oral hygiene home care regimen.

Association between signs of trauma from occlusion and periodontitis.

The purpose of this study was to evaluate the association between signs of trauma from occlusion, severity of periodontitis and radiographic record of bone support. The maxillary first molars of 300 individuals were independently evaluated by two examiners for signs of trauma from occlusion, pattern or occlusal contacts and severity of periodontitis. Each site was also evaluated radiographically by an independent third examiner. The results indicated that: teeth with either bidigital mobility, functional mobility, a widened periodontal ligament space or the presence of radiographically visible calculus had a deeper probing depth, more loss of clinical attachment and less radiographic osseous support than teeth without these findings, teeth with occlusal contacts in centric relation, working, nonworking or protrusive positions did not exhibit any greater severity of periodontitis than teeth without these contacts, teeth with both functional mobility and radiographically widened periodontal ligament space had deeper probing depth, more clinical attachment loss and less radiographic osseous support than teeth without these findings and given equal clinical attachment levels, teeth with evidence of functional mobility and a widened periodontal ligament space had less radiographic osseous support than teeth without these findings.