3rd Injection Research Articles

Efficacy of Intra-Articular Injection of Hyaluronic Acid in the Treatment of Knee Osteoarthritis

This study was conducted to evaluate the efficacy of intraarticular injection of hyaluronic acid for the treatment of knee osteoarthritis. Patients with knee osteoarthritis were followed for a period of six months to assess the efficacy of intra-articular injection of hyaluronic acid given three times in three consecutive weeks. Fifty patients were reviewed at two, eight and 24 weeks post-injection. The average age was 60.9 years and female to male ratio was 3:1. Patients were assessed using the Lequesne Algofunctional Index for function, and the visual analogue score for pain and side effects. We found that the knee pain reduced and the function improved in most patients and these beneficial effects maintain till the last follow up. The only side effect noted was one case of acute non-septic joint effusion after the 3rd injection. We concluded that intra-articular injection of hyaluronic acid can produce pain relief and functional improvement for up to 6 months.

Selective inhibition of forebrain or hindbrain leptin receptors modifies the response to peripheral leptin

Previously we showed that peripheral infusions of physiological doses of leptin increase body fat in chronically decerebrate rats, in which the forebrain is surgically dissociated from the hindbrain. This study tested whether inhibition of leptin receptors (ObR) in the vicinity of either the 3rd or the 4th ventricle modified changes in food intake and body weight caused by peripheral leptin injections. Rats were food deprived during the light cycle and received 3rd ventricle injections of 0, 1.5, 2.5, 3.0 or 3.5 ug of leptin mutein protein (mutein), an ObR antagonist, 30 minutes before an i.p. injection of either PBS or 2.5 mg leptin/kg. Leptin inhibited 36 hour food intake and weight gain in rats receiving 0, 1.5 or 2.5 ug mutein, whereas 3.0 and 3.5 ug mutein increased weight gain and prevented any effect of leptin on either intake or weight gain. In a second study of similar design rats received 4th ventricle injections of 0, 0.75 or 1.5 ug mutein 30 minutes before an i.p. injection of 2.5 mg leptin/kg. Leptin inhibited 24 hour, but not 36 hour, food intake of rats receiving 0 ug mutein. Both doses of mutein inhibited weight gain and food intake and these effects were exaggerated and prolonged by leptin. These observations support those made in decerebrate rats and suggest that ObR in the brainstem oppose the feeding and energy expenditure responses to activation of forebrain ObR. Supported by NIH grant DK053903.

The effect of viscosupplementation on neuromuscular control of the knee in patients with osteoarthritis

The aim of this study was to investigate the short-term effects of intra-articular injection of hyaluronan (Hylan G-F 20) on proprioception, isokinetic muscle force, self reported pain, and functional condition in patients with knee osteoarthritis (OA). 63 patients with stage II-III bilateral knee OA were included in this randomized, placebo controlled, and prospective study. Subjects were randomized with 42 of them into the treatment group and 21 of them into the placebo group. Hyaluronan was intraarticularly injected into both knees of the subjects which were in the treatment group, whereas physiological saline was intraarticularly injected to the subjects which were in the placebo group. Proprioception and the isokinetic muscle force measurement were performed. Visual analogue scale (VAS) and WOMAC scale were used to evaluate pain and physical function. Statistical analysis was performed on 120 knees of 60 patients completing the trial. The average absolute angular error (AAAE) value showing the proprioceptive error level in the treatment group was detected to be statistically significantly lower compared to placebo at the measurements performed after the 3rd injection (p = 0.02) and after one week (p = 0.01). While there was no inter-group difference in isokinetic measurements performed at 180 and 240 degrees/sec, a significant difference was detected at the measurement performed at 60 degrees/sec in favor of the treatment group (p = 0.02). Activity and resting VAS-pain values, WOMAC parameters (except the WOMAC stiffness) were detected to be significantly lower in the treatment group. Local adverse events were not reported in any patient. In this study, it was demonstrated that intraarticular injection of hyaluronan in patients with knee OA led to a short-term increase in proprioception and isokinetic muscle force, and also significant improvements in the functional conditions of patients. Long-term studies are needed.

FIRST VERSUS SECOND EYE INTRAVITREAL RANIBIZUMAB THERAPY FOR WET AMD

To evaluate the short-term visual outcomes after intravitreal ranibizumab for wet age-related macular degeneration, when used in first eyes (good vision in the untreated eye) compared with second eyes (significant visual impairment in the untreated eye). Seventy-five consecutive patients who received intravitreal ranibizumab injection were divided into Group A, comprising 35 first eyed patients and Group B, comprising 40 second eyes. Visual acuity and contrast sensitivity was compared before treatment, and 3 months after the 3rd injection. Results were compared at 95% confidence interval. Mean pretreatment logMar visual acuity was 0.86 (standard deviation 0.28) in Group A whereas Group B was 0.66 (standard deviation 0.36) (P = 0.007). Posttreatment the mean visual acuity in Group A was 0.63 (standard deviation 0.37) and in Group B was 0.44 (standard deviation 0.33) (P = 0.02). The mean numbers of letters gained per patient were 11.1 (Group A) and 10.6 (Group B). Half of all patients showed significant improvement of visual acuity (> or =15 letters gain). Contrast sensitivity significantly improved in both groups and was usually, but not always, associated with visual gain. Second eye patients tend to present to clinical diagnosis at a better visual acuity than first ones and subsequently have better chances for better posttreatment visual acuity. However, both groups have an equal chance of significant visual improvement.

Inhibin A Production after Gonadotropin Stimulus: A New Method to Detect Ovarian Tissue in Ovotesticular Disorder of Sex Development

Background/Aims: While laboratory methods for the detection of testicular tissue are well standardized, currently there is no available test to demonstrate the presence of ovarian tissue. We evaluated the effectiveness of gonadal stimulation with luteinizing hormone (LH)/follicle-stimulating hormone (FSH) for the detection of ovarian tissue in patients with disorders of sex development (DSD). Methods: Ten patients with congenital adrenal hyperplasia (CAH) as ovarian-positive controls, 10 with cryptorchidism (ovarian-negative controls), 13 patients with DSD of no defined etiology and 7 patients with ovotesticular DSD (true hermaphroditism, TH) were included in the study. They underwent a daily injection of both LH and FSH on 3 consecutive days. LH, FSH, estradiol, testosterone and inhibin A were measured before treatment, 24 h after the 1st dose and 24 h after the 3rd dose. Results: Estradiol increased in all CAH and TH patients, with a median value of 155.1 and 92.6 pg/ml, respectively, after the 3rd injection. Inhibin A also increased in all CAH and TH patients, with a median value of 70.4 and 32.2 pg/ml, respectively, after the 3rd injection. There was no change in these hormones in the other groups. Conclusion: The LH/FSH stimulation test might be a useful method to detect the presence of ovarian tissue.

Enhancement of the expression of HCV core gene does not enhance core-specific immune response in DNA immunization: advantages of the heterologous DNA prime, protein boost immunization regimen

BackgroundHepatitis C core protein is an attractive target for HCV vaccine aimed to exterminate HCV infected cells. However, although highly immunogenic in natural infection, core appears to have low immunogenicity in experimental settings. We aimed to design an HCV vaccine prototype based on core, and devise immunization regimens that would lead to potent anti-core immune responses which circumvent the immunogenicity limitations earlier observed.MethodsPlasmids encoding core with no translation initiation signal (pCMVcore); with Kozak sequence (pCMVcoreKozak); and with HCV IRES (pCMVcoreIRES) were designed and expressed in a variety of eukaryotic cells. Polyproteins corresponding to HCV 1b amino acids (aa) 1–98 and 1–173 were expressed in E. coli. C57BL/6 mice were immunized with four 25-μg doses of pCMVcoreKozak, or pCMV (I). BALB/c mice were immunized with 100 μg of either pCMVcore, or pCMVcoreKozak, or pCMVcoreIRES, or empty pCMV (II). Lastly, BALB/c mice were immunized with 20 μg of core aa 1–98 in prime and boost, or with 100 μg of pCMVcoreKozak in prime and 20 μg of core aa 1–98 in boost (III). Antibody response, [3H]-T-incorporation, and cytokine secretion by core/core peptide-stimulated splenocytes were assessed after each immunization.ResultsPlasmids differed in core-expression capacity: mouse fibroblasts transfected with pCMVcore, pCMVcoreIRES and pCMVcoreKozak expressed 0.22 ± 0.18, 0.83 ± 0.5, and 13 ± 5 ng core per cell, respectively. Single immunization with highly expressing pCMVcoreKozak induced specific IFN-γ and IL-2, and weak antibody response. Single immunization with plasmids directing low levels of core expression induced similar levels of cytokines, strong T-cell proliferation (pCMVcoreIRES), and antibodies in titer 103(pCMVcore). Boosting with pCMVcoreKozak induced low antibody response, core-specific T-cell proliferation and IFN-γ secretion that subsided after the 3rd plasmid injection. The latter also led to a decrease in specific IL-2 secretion. The best was the heterologous pCMVcoreKozak prime/protein boost regimen that generated mixed Th1/Th2-cellular response with core-specific antibodies in titer ≥ 3 × 103.ConclusionThus, administration of highly expressed HCV core gene, as one large dose or repeated injections of smaller doses, may suppress core-specific immune response. Instead, the latter is induced by a heterologous DNA prime/protein boost regimen that circumvents the negative effects of intracellular core expression.

TJN-259 Improves Mesangial Lesions in Experimental Immunoglobulin A Nephropathy in ddY Mice

TJN-259 is a chemical substance based on the structural features of the botanically derived ingredient acteoside. This study was performed in order to elucidate the antinephritic effects of TJN-259 in experimental immunoglobulin A (IgA) nephropathy. In this study, 28-week-old ddY mice were used as a spontaneous model of IgA nephropathy. With regard to spontaneous IgA nephropathy, we investigated the effects of TJN-259 administered from 28 to 40 weeks. In addition, an accelerated model of IgA nephropathy was experimentally induced in ddY mice by oral administration of bovine serum albumin, followed by reticuloendothelial blocking by colloidal carbon injection and heminephrectomy. At 10 weeks after the 3rd carbon injection, we also examined the effects of TJN-259 on accelerated IgA nephropathy. To investigate the effects of TJN-259 on transforming growth factor (TGF)-beta1 production in accelerated IgA nephropathy, kidneys were isolated and measured TGF-beta1 by the enzyme-linked immunosorbent assay (ELISA) method. The administration of TJN-259 to mice with spontaneous IgA nephropathy decreased the incidence of mesangial expansion as well as the number of nuclei per glomerular cross-section in comparison with that of non-treated mice. In addition, TJN-259 treatment prevented the increase in the incidence of mesangial expansion, crescent formation, and segmental sclerosis in glomeruli in accelerated IgA nephropathy. TJN-259 also inhibited the increased immunostaining score of collagen type IV and TGF-beta1 in glomeruli of accelerated IgA nephropathy. Treatment with TJN-259 inhibited the increases in renal total and mature TGF-beta1 protein levels in accelerated type IgA nephropathy. TJN-259 failed to inhibit the increase in serum IgA levels in both models. These results suggest that TJN-259 was an effective treatment against IgA nephropathy in ddY mice, acting via the suppression of TGF-beta1 production in glomeruli.

Phase 1 Clinical Trial of NPI-0052, a Novel Proteasome Inhibitor in Patients with Multiple Myeloma

Background: NPI-0052 is a novel proteasome inhibitor that produces prolonged inhibition of all three catalytic activities (C-L, T-L, CT-L) of the 20S proteasome. Preclinical data suggest NPI-0052 may demonstrate an improved therapeutic ratio, with significant activity in hematologic and solid tumor malignancies including multiple myeloma (MM) resistant to bortezomib and other agents (Chauhan et al, Blood 2006). Studies have therefore been initiated in patients with multiple myeloma, lymphoma, leukemia and solid tumors.Materials and Methods: This Phase 1 dose escalation study evaluated NPI-0052 monotherapy in patients with relapsed and relapsed/refractory MM, including those that have received bortezomib and/or lenalidomide. Patients were assessed for safety, pharmacodynamics (including ex vivo proteasome inhibition), plasma pharmacokinetics (PK), and clinical activity (with response assessed by modified EBMT criteria). Patients were treated with NPI-0052 administered as a weekly IV injection on Days 1, 8 and 15 every 4 weeks with concomitant hydration. The dose of NPI-0052 was escalated in cohorts of 3 patients dependent on observed adverse events utilizing a 3+3 design. Proteasome inhibition and PK were assayed after the 1st and 3rd dose and upon any intra-patient dose escalation. Proteasome inhibition was also assessed in CD138 positive cells isolated from bone marrow aspirates obtained at baseline and after the 3rd dose in a subset of patients.Preliminary Results: To date, 10 patients have been treated at doses ranging from 0.025 mg/m2 to 0.075 mg/m2 without reaching an MTD. One patient experienced reversible elevation in serum creatinine that responded to drug cessation and steroids; this event may have been related to progression of his underlying light chain nephropathy (as interval worsening of renal function was noted prior to enrollment). Drug-related adverse events have been otherwise unremarkable at all dose levels tested. PK data demonstrate a rapid elimination half life (estimated to be 3–4 minutes) with clearance between 8–21 mL/min and Vz of 44–99L; no change in PK has been observed comparing the 1st and 3rd injection. Proteasome inhibition in whole blood suggests drug-dependent CT-L inhibition, with inhibition up to 28% observed (inhibition up to 100% at doses of up to 0.7 mg/m2 has been observed in other clinical trials with NPI-0052 without producing the profile of toxicity reported with standard doses of bortezomib). Whilst no responses have been confirmed, two patients with relapse/refractory MM remained on study for over 6 months and one year, respectively, with stable disease and no significant toxicity. Importantly, no peripheral neuropathy or myelosuppression has been seen in 41 treatment cycles in patients to date.Conclusions: In patients with relapsed and relapsed, refractory MM, NPI-0052 affects parameters relevant to pharmacodynamics, PK and demonstrated potential clinical benefit at doses well below the MTD anticipated from Phase 1 clinical trials with NPI-0052 in lymphoma and solid tumors. Drug administration to date has been well tolerated. Dose escalation continues to define DLT and MTD, and to recommend a phase 2 dose for further study in patients with advanced MM.

The effects of acute multiple intraperitoneal injections of the GABA B receptor agonist baclofen on food intake in rats

This study was undertaken to examine the effects of acute repeated administration of the GABA B receptor agonist baclofen on food intake in rats. In Experiment 1, the effects of repeated intraperitoneal (i.p.) injections of the GABA B receptor agonist baclofen (1 and 2 mg/kg) at 2 h intervals were investigated on food intake in non-deprived male Wistar rats. Both doses of baclofen significantly increased food intake after the 1st injection ( P < 0.05), but had no effects on intake following the 2nd and 3rd injections. By contrast, in Experiment 2, diazepam (1 and 2 mg/kg, i.p.) significantly increased food intake (at least, P < 0.05) after each of 3 injection separated by 2 h in non-deprived rats. These data show that tolerance occurs to the hyperphagic effects of baclofen with acute multiple injections, and may have important implications for future studies investigating the effects of GABA B receptor agonists on food intake and energy homeostasis.

Botulinum toxin type A (Botox) versus botulinum toxin type B (Myobloc) in subjects with cervical dystonia: A comparison of the frequency of xerostomia and dysphagia (AN072-402 EU/CD)

Objective : To evaluate the effect of switching BoNT serotypes on occurrence of xerostomia/dysphagia. Methods : In a double-blind (DB) study, BoNT-naive CD subjects were randomized to receive either 2 consecutive BoNTA doses (150 U/200 U) or 2 consecutive BoNTB doses(10k U/15k U). Subjects then enrolled in an open-label (OL) study and >3 months later received 10k BoNTB as their 1st OL injection (3rd BoNT injection). The incidence of xerostomia/dysphagia following the 1st OL treatment was compared in subjects originally treated with BoNTA vs. BoNTB treated in the DB phase. Xerostomia/dysphagia reports after the 1st BoNTB injection (1st OL injection for BoNTA DB subjects and 1st DB injection for BoNTB DB subjects) were also compared. Results : Subjects receiving BoNTA in the DB phase experienced significantly more xerostomia (31% vs. 11%, respect.; p =0.025) and dysphagia (33% vs. 9%, respect.; p =0.006) following their 1st OL BoNTB treatment than those who were previously exposed to BoNTB in the DB phase. The incidence of xerostomia (31% vs. 39%, respect.; p =0.415) and dysphagia (33% vs. 9%, respect.; p =0.067) after the initial exposure to BoNTB was similar for the group 1st randomized to BoNTA in the DB phase (AE occurrence in 1st OL treatment) vs. the group 1st randomized to BoNTB in the DB phase (AE occurrence in 1st DB treatment). Conclusions : Although tolerance to xerostomia/dysphagia with repeating BoNTB doses has been demonstrated, no evidence exists of cross-tolerance between BoNTA and BoNTB for these AE.

Evaluation of the effect of a 3rd GnRH injection administered six days after the 2nd GnRH injection of Ovsynch on the reproductive performance of Japanese black cows

This study was designed to evaluate the reproductive performance of Japanese black cows following the 3rd injection of gonadotropin releasing hormone (GnRH) analogue administered concurrently with Ovsynch-based treatment on day 6 (day 1 = the day of ovulation). In Experiment 1, 12 cows were allocated into three groups: a control group that was subjected to Ovsynch treatment and then injected with a placebo on day 6; group 1 (Ovsynch + GnRH), which was subjected to Ovsynch treatment and was injected with GnRH analogue on day 6, and group 2 (Ovsynch + controlled internal drug-release (CIDR) + GnRH), which received Ovsynch-CIDR treatment and was injected with GnRH analogue on day 6. Blood collection and ultrasonographic observation of the ovaries were conducted daily. Both treatments induced the formation of an accessory corpus luteum and significantly increased the cross-sectional area of the luteal tissue when compared to the control. However, plasma progesterone (P4) was significantly higher in the treatment groups than in the control group on days 11, 12, 17 and 18 in the group 1 and from day 10 to 21 in the group 2. In Experiment 2, 41 cows were assigned to the same three groups described above and then artificially inseminated on day 1. The pregnancy rates on day 45 did not differ among groups. In conclusion, administration of GnRH analogue on day 6 following Ovsynch-based treatment did not improve the reproductive performance of Japanese black cows, even though the P4 concentration was higher in groups that received the GnRH.

Development of a Novel Mouse Anaphylaxis Model Produced by Intermittent Intravenous Injections of Ovalbumin without Adjuvant

This study was performed to develop mouse anaphylaxis model using ovalbumin (OA) as an antigen, with no adjuvant, and an intravenous route of administration. OA at dose levels from 0.5 to 500 mg/kg was injected 3 times into the tail vein of female CD-1 (ICR) mice at 4-day intervals. Clinical signs and extravasation of Evans blue dye (vascular permeability) were observed, serum histamine levels and airway resistance were measured, and histopathological examinations were performed. Furthermore, to detect serum IgE and IgG, heterologous and homologous passive cutaneous anaphylaxis tests were performed. Additionally, strain differences in susceptibility to OA anaphylaxis was examined with 3 mouse strains: ICR, BALB/c and C57BL/6 mice. Shortly after the 3rd injection, most of the animals exhibited systemic anaphylactic signs including death, and this was accompanied by increases in vascular permeability, plasma histamine levels, and airway resistance. Four days after the 2nd injection, anti-OA IgE and IgG were detected in sera of mice injected with 0.5 and 5.0 mg/kg or higher doses of OA, respectively. Histological findings included: congestion in many tissues; detachment of mucosal epithelial cells in the gastrointestinal tract; enhanced death, decreased numbers of lymphocytes, and increased numbers of macrophages in the lymphoid tissues. ICR mice were shown to be most susceptible to the OA anaphylaxis of the 3 mouse strains examined. In conclusion, the results demonstrate that 3 intermittent intravenous injections of OA without adjuvant at 4-day intervals can induce anaphylactic shock in mice, in particular, most effectively in ICR mice, and that the anaphylaxis is characterized by specific IgE and IgG production.

Mediation of burn-induced hypermetabolism by CRF receptor-2 activity

Hypermetabolism and anorexia are significant problems associated with major burn trauma. Recent studies have implicated hypothalamic peptides and receptors of the corticotropin releasing factor (CRF) family as putative mediators of burn-induced hypermetabolism. Increased neuronal activity at the CRF type 2 receptor (CRF R-2) appeared particularly involved in the expression of elevated resting energy expenditure (REE) following major burn trauma. In the present study we continued these investigations of CRF R-2 mediation of burn-induced hypermetabolism, demonstrating that 3rd ventricle injection of CRF R-2 antisense oligodeoxynucleotide (ODN) normalized REE in burned rats. Similar treatments with CRF or CRF R-1 antisense ODNs had no significant effect in burned rats. In addition, 3rd ventricle injection of the selective CRF R-2 antagonist, antisauvagine-30, also reduced REE significantly in burned rats, while similar treatment with the selective CRF R-1 antagonist, antalarmin, was without effect. To determine which endogenous peptide was altered following burn we measured hypothalamic levels of urocortin (UCN) and CRF 15 days after burn injury, finding UCN was significantly elevated by nearly 3-fold, while CRF level tended to be decreased. We also assessed hypothalamic mRNA peptide and receptor expression by real-time PCR 7, 14, and 21 days post-burn, observing decreased CRF expression 7 and 21 days post-burn, decreased UCN-2 expression 7 days post-burn, and no significant alteration in UCN-1 at any time point. However, CRF R-2 mRNA was elevated at each post-burn time point. These results continue to suggest that increased neuronal activity is integrally involved in the mediation of burn-induced hypermetabolism, and that one of the UCN peptides may be the endogenous ligand affecting this receptor.

Deficiency of tenascin‐C attenuates liver fibrosis in immune‐mediated chronic hepatitis in mice

Tenascin-C (TNC), an extracellular matrix glycoprotein, is upregulated in chronic liver disease. Here, we investigated the contribution of TNC to liver fibrogenesis by comparing immune-mediated hepatitis in wild-type (WT) and TNC-null (TNKO) mice. Eight-week-old BALB/c mice received weekly intravenous injections of concanavalin A to induce hepatitis, and were sacrificed one week after the 3rd, 6th, 9th, and 12th injections. In WT livers, immunohistochemical staining revealed a gradual increase in TNC deposition. TNC mRNA levels also increased sequentially and peaked after the 9th injection. Collagen deposition, stained with picrosirius red, was significantly less intense in TNKO mice than in WT mice, and procollagen I and III transcripts were significantly upregulated in WT mice compared with TNKO mice. Inflammatory infiltrates were most prominent after the 3rd-6th injections in both groups and were less intense in TNKO mice than in WT mice. Interferon-gamma, tumour necrosis factor-alpha, and interleukin-4 mRNA levels were significantly higher in WT mice than in TNKO mice, while activated hepatic stellate cells (HSCs) and myofibroblasts, a cellular source of TNC and procollagens, were more common in WT livers. Transforming growth factor (TGF)-beta1 mRNA expression was significantly upregulated in WT mice, but not in TNKO mice. In conclusion, TNC can promote liver fibrogenesis through enhancement of inflammatory response with cytokine upregulation, HSC recruitment, and TGF-beta expression during progression of hepatitis to fibrosis.

Possible role of CRF peptides in burn-induced hypermetabolism

Hypermetabolism and anorexia are significant problems associated with major burn trauma. Recent studies have shown that hypothalamic corticotropin releasing factor (CRF) elevates metabolic rate, while neuropeptide Y (NPY) reduces it. CRF also elicits anorexia, while NPY stimulates feeding. We hypothesized that elevation of CRF and decrease of NPY may be mediators of these negative effects of burn trauma. Therefore, we assessed concentrations of CRF and NPY in hypothalamus of burned rats one, three, and twenty-one days after a 30% body surface area, full-thickness, open flame burn. In addition we determined whether a biochemical lesion of CRF receptors using 3rd ventricle injections of a saporin–CRF conjugated peptide would decrease resting energy expenditure (REE). We found a three-day period of anorexia, with REE significantly increasing three days after the burn trauma. Concentrations of NPY were increased in the PVN-containing dorsomedial region of the hypothalamus 1 and 3 days after burn trauma, but were increased further in the day 1 pair-fed rats suggesting this change was a consequence of the anorexia. Levels of CRF were decreased in the ventromedial region of the hypothalamus in day 1 and day 3 burned and PF rats. Treatment with the saporin–CRF conjugate normalized REE and reduced CRF receptor-2 density in the hypothalamus of burned rats, and blocked CRF-induced hypermetabolism in sham-burned rats. Although these results suggest a role of CRF receptors in mediating burn-induced hypermetabolism, CRF itself may not be the principle ligand, as suggested by the significant elevation of hypothalamic urocortin 15 days after burn injury.

IgE Production after Four Routes of Injections of Japanese Cedar Pollen Allergen without Adjuvant: Crucial Role of Resident Cells at Intraperitoneal or Intranasal Injection Site in the Production of Specific IgE toward the Allergen

The production of specific IgE antibodies directed toward cedar pollen correlates well with the onset of allergic rhinitis; but the mechanisms of allergen recognition as nonself and Ig class switch to IgE by the immune system are still not fully understood. In the present study, we injected cedar pollen into mice through 4 different routes (intranasal (i.n.), intraperitoneal (i.p.), intravenous (i.v.), and subcutaneous (s.c.)) without adjuvant 1 to 3 times, and determined time-dependent changes in the total and specific serum IgE levels compared with those in the serum levels of other isotype Igs. After an i.p. or i.n. injection of allergen into the mice, they produced a 1.5-to 1.7-fold increase in total IgE, but none in IgG, IgM, or IgA antibodies in their serum, whereas an i.v. or s.c. injection of allergen was inactive as an inducer of total IgE antibodies. Upon a 2nd (s.c.) injection of the allergen into the i.p. or i.n. sensitized mice, a large amount of allergen-specific IgE antibodies was found in the serum. In the case of i.v. or s.c. sensitized mice, however, they produced total, but not specific, IgE antibodies; and a 3rd (s.c.) injection of the allergen resulted in a large amount of specific IgE antibodies in the serum. These results imply that resident cells at the i.p. or i.n. injection site may play a crucial role in the efficient production of total and specific IgE antibodies toward the allergen.

Up-regulation of adenosine A 1 receptor binding in pentylenetetrazol kindling in mice: effects of angiotensin IV

The effects of the hexapeptide angiotensin II (3–8) ANG IV, the selective A 1 receptor agonist cyclohexyladenosine (CHA) and the combination of ANG IV + CHA on pentylenetetrazol (PTZ)-generalized seizures; kindling development and maintenance were studied. By using in vitro quantitative receptor autoradiography, the regulation of adenosine A 1 receptor density at different time points during the kindling procedure and postkindling period was determined. ANG IV and CHA effectively reduced clonic seizures in PTZ-generalized seizure model, in PTZ-kindled mice as well as during kindling development and a week later by rechallenge with PTZ. Furthermore, coadministration of ANG IV and CHA had a strong anticonvulsant effect, both compounds acting synergistically. A significant increase of adenosine A 1 receptor density was detected in somatosensory cortex, hippocampus, amygdala and geniculate nuclei early in the kindling procedure (after the 3rd injection), which persisted at least 1 month after the end of kindling procedure. In addition, a delayed up-regulation of adenosine A 1 receptor binding was observed a week after kindling in the mamillary bodies and a month later in the motor cortex. The pretreatment with ANG IV caused a down-regulation of adenosine A 1 receptor density to the control level in most time points and brain areas. In conclusion, PTZ kindling-induced increase of adenosine A 1 receptor binding at different time points and in specific brain structures might represent an adaptive mechanism for coping with the hyperexcitability typical for this phenomenon. The antiepileptogenic effect of ANG IV could be realized partly through an adenosine-dependent mechanism.

Voltage Waveforms Comparison for Different PWM Modulation Strategies

A comparison between several commonly used PWM modulation strategies is made, namely Sinusoidal Regular-Sampled Asymmetrical PWM, Regular-Sampled Asymmetrical PWM plus 3rd harmonic injection and Centred Asymmetrical Space Vector PWM, in order to determine which of them leads to a higher voltage waveform quality. It is shown that the first PWM method causes higher harmonic losses and that although 3rd harmonic injection PWM spectrum contains few harmonic terms than Centred Asymmetrical Space Vector PWM, the latter shows less weighted harmonic distortion.

Animal Models of Chronic Pain: Gene Therapy using naked plasmid DNA coding the anti-inflammatory gene, Interleukin-10 (IL10): repeated injections lead to long-term pain control in chronic constriction injury (CCI) rats

The proinflammatoy cytokine, interleukin-1 [IL1], released from activated spinal cord glia (astrocytes & microglia) has been shown to critically mediate the maintenance of pathological pain produced by Chronic Constriction Injury (CCI). The anti-inflammatory cytokine, IL10, suppresses the production & function of the proinflammatory cytokines IL1, tumor necrosis factor [TNF] & interleukin-6 [IL6]). Here, we used spinal delivery (intrathecal; i.t.) of naked plasmid DNA coding the rat IL10 gene (pDNA-IL10) or the reporter gene, jellyfish green fluorescent protein (control; pDNA-GFP) to (a) reverse exaggerated pain after single & multiple injections, (b) measure pDNA-IL10 induced changes of proinflammatory cytokine & IL10 protein & mRNA levels in spinal tissue and/or release into lumbosacral & cervical cerebrospinal fluid (CSF), & (c) to visualize pDNA-GFP transfected cells . We tested CCI-induced thermal hyperalgesia and allodynia. Following behavioral testing, lumbar dorsal cord & cerebrospinal fluid (CSF) were assayed. Behavioral measures (von Frey test and/or thermal Hargreaves test) were assessed before, 10 days after CCI & every 1-2 days after i.t. pDNA for 70 days. The time interval that i.t. pDNA-IL10 reversed CCI-induced allodynia & hyperalgesia increased following each injection. Allodynia was stably reversed for 3 weeks after the 3rd injection. Following 2 pDNA-10 injections in behaviorally verified rats, i.t. pDNA-IL10 increased human IL10 & decreased rat IL1protein released into CSF; no changes were observed in spinal tissue. Spinal cauda equina IL10 mRNA levels are altered by i.t. pDNA-IL10 vs. pDNA-GFP. Bright GFP was observed (autofluorescence) in ependymal cells of the cauda equina from pDNA-GFP treated rats vs. controls. Ongoing studies are examining 1) spinal changes in mRNA for TNF, IL6 & IL10 &amp;/or their receptors, & 2) whether IL10 facilitates gene transcription. MH01558, MH00314, NS38020, Juvenile Diab. Res. Found. & NIH DK58327, HL59412.

Graphical and Algebraic Synthesis for PWM Methods

The article proposes an algebraic synthesis for PWM methods, which is based on a graphical representation of the 2 levels VSI (Voltage Source Inverter). This theory can be the starting point for every PWM method representation.In order to understand the historical evolution of the PWM methods the authors propose two classification groups of the main PWM techniques. The main classification is based on the movement of the neutral point (zero-voltage VN0 evolution). The details of PWM methods presented here are also seen from the graphical representation perspective.The equivalence between the well-known SVM (Space Vector Modulation) practically DDT realized (Direct Digital Technique) and the three-phase PWM with a carrier based industrial realization is shown. The principles of the three-phase PWM and the 3rd harmonic injection PWM (THIPWM4, THIPWM6) are compared the same as the DPWM (Discontinuous PWM) methods. These classifications and equivalences between different techniques show that the number of contemporary used PWM principles can be reduced. The general complex space representation and the classifications proposed here are simple to comprehension educational tools and they simply the way towards practical implementation.