3rd Injection Research Articles

Comparative analysis of immunization schedules using a novel adenovirus-based immunotherapeutic targeting hepatitis B in naïve and tolerant mouse models

Development of active targeted immunotherapeutics is a rapid developing field in the arena of chronic infectious diseases. The question of repeated, closely spaced administration of immunotherapeutics to achieve a rapid impact on the replicating agent is an important one. We analyzed here, using a prototype adenovirus-based immunotherapeutic encoding Core and Polymerase from the hepatitis B virus (Ad-HBV), the influence of closely spaced repeated immunizations on the level and quality of induced HBV-specific and vector-specific immune responses in various mouse models. Ad-HBV, whether injected once or multiple times, was able to induce HBV- and adeno-specific T cells both in HBV-free mice and in a HBV tolerant mouse model. Adenovirus-specific T cell responses and titers of neutralizing anti-Ad5 antibodies increased from time of the 3rd injection. Interestingly, single or multiple Ad-HBV injections resulted in detection of Polymerase-specific functional T cells in HBV tolerant mice. Overall no modulation of the levels of HBV-specific cytokine-producing (IFNγ/TNFα) and cytolytic T cells was observed following repeated administrations (3 or 6 weekly injections) when compared with levels detected after a single injection with the exception of two markers: 1. the proportion of HBV-specific IFNγ-producing cells bearing the CD27+/CD43+ phenotype appeared to be sustained in C57BL/6J mice following 6 weekly injections; 2. the percentage of IFNγ/TNFα Core-specific producing cells observed in spleens of HLA-A2 mice as well as of that specific of Polymerase observed in livers of HBV tolerant mice was maintained. In addition, percentage of HBV-specific T cells expressing PD-1 was not increased by multiple injections. Overall these data show that, under experimental conditions used, rapid, closely spaced administrations of an adenovirus-based HBV immunotherapeutics does not inhibit induced T-cell responses including in a HBV-tolerant environment.

A98: Rescue Treatment by Increased Doses of IL‐1 Inhibitors for Macrophage Activation Syndrome in Children With Systemic Juvenile Idiopathic Arthritis

Background/Purpose:macrophage activation syndrome (MAS) is a life‐threatening complication of systemic juvenile idiopathic arthritis (SJIA). More than half cases of MAS are steroid‐resistant and required to use second‐line agents, such as cyclosporine A (CsA), intravenous immunoglobulin (IVIG) and biologic agents. Known interleukin‐1 (IL‐1) is the key cytokine in pathogenesis of MAS and SJIA, so anti‐IL‐1 treatment considered as potential effective treatment. There are several publication about efficacy of increased doses of anakinra during the MAS episodes. Data about efficacy others new IL‐1 blockers (canakinumab and rilonacept) for MAS in SJIA are limited. There are only several cases of MAS during and after canakinumab (CNKB) treatment of SJIA in randomized controlled trials.Methods:a retrospective study of the clinical notes, reports of laboratory investigations and radiological images. We included 3 patients with SJIA who developed MAS. First patient (Pt#1) had the severe form of MAS, resistant to high doses of steroids, IVIG, CsA. After failure of previous treatment anakinra was added, and later changed on CNKB. Second patient (Pt#2) developed SJIA flare with severe MAS after 3rd CNKB injection, resistant to high doses steroids, IVIG, required Fresh Frozen Plasma, third patient (Pt#3) developed only mild MAS after 8th CNKB injection without signs of SJIA flare. Indications for increased doses of CNKB were inefficacy of previous anti IL‐1 treatment in standard and increased doses and severity of MAS (Pt#1, Pt#2) according the manufacture's manual permitted to use the maximum dose of 300 mg per injection. The initial CNKB dose before MAS was 4 mg/kg every 4 weeks in Pt#2 and Pt#3. All patient during the MAS received one single injection of increased dose of CNKB: 15 mg/kg (Pt#1), 12.5 mg/kg (Pt#2) and 7.5 mg/kg (Pt#3). CNKB was injected every 4 weeks with gradually tapering dose up to 4 mg/kg every 4 weeks. All parents signed the inform consent. Institutional Review Board affirmed this kind of treatment.Results:in Pt#1 initial treatment with standard doses of anakinra was insufficient and dose was increased up to 20 mg/kg/day with good response. Later anakinra was changed on CNKB in 1st dose 15 mg/kg every 4 weeks with gradually tapering dose up to 6 mg/kg up to now. The switching from anakinra to CNKB did not loose the efficacy. In Pt#2 after initiation of increased dose of CNKB the flare of JIA and active MAS was dramatically abrupt up to 7th day after 1st injection. Total normalization of lab tests occurred in 2 weeks. After normalization we tapered corticosteroids. In Pt#3 MAS also quickly disappeared, the normalization of lab test also occurred in 10 days. No side and adverse effects were noticed during using of increased anakinra and CNKB doses.Conclusion:SJIA and MAS are both IL‐1‐ dependent conditions. The SJIA flare and MAS usually associated with increased production of pro‐inflammatory cytokines, especially IL‐1β. So we suppose that increased levels of IL‐1β require increased doses of IL‐1 blockers. Our data shows the efficacy and safety of increased doses of IL‐1 blockers. It is necessary to provide future trials to evaluate the efficacy and safety increased doses of IL‐1 blockers for treatment MAS in SJIA.

Aqueous Interleukin-6 (IL-6) Level Is a Marker for Treatment Resistance to Bevacizumab in Age-Related Macular Degeneration —Aqueous Cytokines after Bevacizumab

Background: To prospectively evaluate the effect of intravitreal bevacizumab on aqueous levels of interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF) in patients with exudative age-related macular degeneration (AMD) and to correlate clinical outcomes of patients and aqueous cytokine levels before and after injection. Methods: The study group consisted of 30 eyes from 30 patients with exudative AMD who underwent intravitreal injection of bevacizumab three times at monthly intervals. The aqueous samples prior to the 1st injection (baseline) and 3rd injection were analyzed for VEGF and IL-6 levels, evaluating the effect of 2 doses of intravitreal bevacizumab. Study patients were sub-grouped based upon change in central subfield (CSF) macular thickness on SD-OCT, at 8 weeks. Group 1 included patients (n = 14) with a decrease in CSF thickness greater than 10% from the baseline and were categorized to have “improved”. Group 2 included patients (n = 16) who had a decrease in CSF thickness 10% or less and were considered “treatment-resistant”. Results: There was no statistically significant change in aqueous VEGF and IL-6 levels after intravitreal bevacizumab. In sub-group analysis, in both Groups 1 and 2 patients, aqueous IL-6 levels showed a better correlation with CSF thickness on SD-OCT (r = 0.72 and 0.71, respectively). Conclusions: Data from our study suggest that aqueous IL-6 may be an important marker of treatment response or resistance. Future therapeutic strategies may include targeted treatment against both VEGF and IL-6, in patients who do not respond to anti-VEGF treatment alone.

Aqueous interleukin-6 levels are superior to vascular endothelial growth factor in predicting therapeutic response to bevacizumab in age-related macular degeneration.

Objective. To prospectively evaluate the effect of intravitreal bevacizumab on aqueous levels of interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF) in patients with exudative age-related macular degeneration (AMD) and correlate clinical outcomes with cytokine levels. Methods. 30 eyes of 30 patients with exudative AMD underwent intravitreal injection of bevacizumab three times at monthly intervals. The aqueous samples prior to the 1st injection (baseline) and 3rd injection were analyzed for VEGF and IL-6 levels. Subjects were subgrouped based upon change in the central subfield (CSF) macular thickness on SD-OCT at 8 weeks. Group 1 included patients (n = 14) with a decrease in CSF thickness greater than 10% from the baseline (improved group). Group 2 included patients (n = 16) who had a decrease in CSF thickness 10% or less (treatment-resistant). Results. In subgroup analysis, in both groups 1 and 2 patients, compared to aqueous VEGF, aqueous IL-6 levels showed a better correlation with CSF thickness on SD-OCT (r = 0.72 and 0.71, resp.). Conclusions. Aqueous IL-6 may be an important marker of treatment response or resistance in wet macular degeneration. Future therapeutic strategies may include targeted treatment against both VEGF and IL-6, in patients who do not respond to anti-VEGF treatment alone.

English

OBJECTIVE: To evaluate the safety and efficacy of intralesional triamcinolone acetonide (TA) injection in primary chalazion. DESIGN: Prospective, consecutive case series. MATERIALS AND METHOD: A prospective study of sixty six(66) patients with primary chalazion presenting to outpatient clinic of Department of Ophthalmology at Tripura medical college & Dr. B.R. Ambedkar Teaching Hospital, Agartala from 1st December 2012 to 3oth September 2013. All patients received intralesional injection of 0.2ml TA (40mg/ml). Data regarding size including digital colour photography, lesion regression or recurrence, and complete ophthalmic examination, were recorded at the time of injection and at different intervals until resolution or surgical excision. Success was defined as at least 80% decrease in size with no recurrence. If the lesion recurred or regression was minimal (<50%), further injections were given as needed. Patients who did not respond to 3rd injections were referred for surgical incision and curettage. RESULT: Around 80% decrease in size of the lesion was noted in forty-eight (48) patients after 1st injection (72.7%). Twelve (12) patients showed complete resolution of the lesion with 2nd injection (18.17%). 3rd injection was needed only in two (2) patients for satisfactory outcome. Four (4) patients were referred for incision and curettage that didn’t show positive response after 3rd injection also. Intraocular pressure and visual acuity remained stable after treatment. One patient developed depigmentation of the skin at the site of injection. No other major complications such as visual loss, subcutaneous fat atrophy were noted with steroid injections. CONCLUSION: Intralesional triamcinolone acetonide (TA) injection is a safe, effective and a simple method of treating primary chalazion where diagnosis is straightforward.

Primary effects of intravitreal bevacizumab inpatients with diabetic macular edema

To evaluate the efficacy of primary intra vitreal bevacizumab (IVB) injection on macular edema in diabetic patients with improvement in best corrected visual acuity (BCVA) and central macular thickness (CMT) on optical coherence tomography (OCT). This prospective interventional case series study was conducted at Retina Clinic, Al-Ibrahim Eye Hospital, and Isra Postgraduate Institute of Ophthalmology Karachi. Between December 2010 to June 2012. BCVA measurement with Early Treatment in Diabetic Retinopathy Study (ETDRS) charts and ophthalmic examination, including Slit-lamp bio microscopy, indirect ophthalmoscopy, Fundus fluorescein angiography (FFA) and OCT were done at the base line examination. At monthly interval all patients were treated with 3 injections of 0.05 ml intra vitreal injection containing 1.25 mg bevacizumab. Patients were followed up for 6 months and BCVA and OCT were taken at the final visit at 6 month. The mean BCVA at base line was 0.42±0.14 Log Mar units. This improved to 0.34±0.13, 0.25±0.12, 0.17±0.12 and 0.16±0.14 Log Mar units at 1 month after 1(st), 2(nd) 3(rd) injections and at final visit at 6 months respectively, a difference that was statistically significant (P>0.0001) from base line. The mean 1mm CMT measurement was 452.9 ± 143.1 µm at base line, improving to 279.8 ± 65.2 µm (P<0.0001) on final visit. No serious complications were observed. Primary IVB at a dose of 1.25 mg on monthly interval seems to provide stability and improvement in BCVA and CMT in patient with DME.

AB0985 Comparison of two different intra-articular hyaluronic acid compounds for carpo-metacarpal joint osteoartrhitis

BackgroundCarpometacarpal osteoarthritis (CMC OA) is highly prevalent in older adults, and is often unresponsive to medical treatment. Intra-articular hyaluronic acid (HA) has been widely shown to improve pain and function...

Efficacy Of Boldenone Undecylenate And Probiotic As Growth Promotors On Male Buffalo Calves

This study was planned to investigate the effect of probiotic and Boldenone undecylenate on body weight gain, feed conversion rate, blood picture and some biochemical alterations of male buffalo calves. This study was done on twenty male healthy buffalo calves 7-8 months old and weight ranges from 164-170 Kg. They were divided into 4 equal groups each of 5 calves, 1st group healthy buffalo calves (control), 2nd group healthy buffalo calves treated with probiotic (Biogen), 3rd group, healthy buffalo calves treated with half therapeutic dose of Boldenone undecylenate and 4th group healthy buffalo calves treated with the therapeutic dose of Boldenone undecylenate. Two blood samples were collected at 1st, 2nd, 3rd, 4th and 5th injection at 4 weeks and 4 weeks post last injection. 1st Sample used for hematological studies. 2nd sample used for biochemical analysis. Oral dosing of probiotic and I/M injection of full dose of boldenone undecylenate induced a significant increase in weight gain, erythrocytic count, hemoglobin% content, packed cell volume %, total leukocytic count, total proteins, , globulin, calcium and phosphorus and significant decrease in feed conversion rate beside insignificant effect on albumin compared with control buffalo calves. Half therapeutic dose of boldenone undecylenate induce insignificant effect in all above parameters. Probiotic and Boldenone undecylenate in therapeutic dose showed non significant effect on serum ALT, AST, ALP, urea and creatinine and significant decrease in blood glucose level. It could be concluded that the Probiotic and boldenone undecylenate induce good improvement in body weight and feed conversion rate but boldenone undecylenate has some adverse effects on some biochemical parameters than probiotic, so probiotics can be used as safe growth promoter.

Electrical stimulation of left anterior thalamic nucleus with high-frequency and low-intensity currents reduces the rate of pilocarpine-induced epilepsy in rats

PurposeBilateral electrical stimulation of anterior nuclei of thalamus (ANT) has shown promising effects on epileptic seizures. However, bilateral implantation increases the risk of surgical complications and side effects. This study was undertaken to access the effectiveness of a stimulation paradigm involving high frequency and low intensity currents to stimulate the left ANT in rats. MethodsMale Sprague-Dawley rats were implanted with electroencephalogram (EEG) electrodes, and an additional concentric bipolar stimulation electrode into either the left or right ANT. The stimulus was a train of pulses (90μs duration each) delivered with a frequency of 200Hz and a current intensity of 50μA. Thalamic stimuli were started 1h before the first intraperitoneal pilocarpine injection (i.p., 300mg/kg), and were applied for 5h. ResultsEEG documented seizure activity and status epilepticus (SE) developed in 87.5% of rats treated with no ANT stimulation after a single dose of pilocarpine. Left ANT stimulation significantly increased the tolerance threshold for pilocarpine-induced EEG seizure activity; 20% of rats developed their EEG documented seizure activity after receiving the first dose, whereas 50%, 10% and 20% of rats did not develop seizure activity until they had received the 2nd, 3rd and 4th pilocarpine injection at 1-h intervals. Moreover, left thalamic stimulation reduced the occurrences of both EEG documented seizure activity and SE induced by single-dose pilocarpine to 25%. However, our result demonstrated that little effect on the occurrence rate of seizures and SE was found when rats received right ANT stimulation. ConclusionsThese results suggest that continuously 5-h left ANT stimulation with high frequency and low intensity currents, beginning from 1h before the pilocarpine administration, may successfully reduce the occurrence rate of EEG documented seizure activity and SE development in rats.

Cervical Myelomalacia in a Lamb (Ovis aries) and an Alpaca (Vicugna pacos) after Attempted Intramuscular Injection

Cervical Myelomalacia in a Lamb (<i><scp>O</scp>vis aries</i>) and an Alpaca (<i><scp>V</scp>icugna pacos</i>) after Attempted Intramuscular Injection

Nesfatin-130–59 but not the N- and C-terminal fragments, nesfatin-11–29 and nesfatin-160–82 injected intracerebroventricularly decreases dark phase food intake by increasing inter-meal intervals in mice

Nesfatin-1 is an 82 amino acid N-terminal fragment of nucleobindin2 that was consistently shown to reduce dark phase food intake upon brain injection in rodents. We recently reported that nesfatin-11–82 injected intracerebroventricularly (icv) reduces dark phase feeding in mice. Moreover, intraperitoneal injection of mid-fragment nesfatin-1 (nesfatin-130–59) mimics the food intake-reducing effects of nesfatin-11–82, whereas N-terminal (nesfatin-11–29) and C-terminal fragments (nesfatin-160–82) did not. We therefore characterized the structure-activity relationship of nesfatin-1 injected icv to influence the dark phase meal pattern in mice. Mouse nesfatin-11–29, nesfatin-130–59, nesfatin-160–82 or vehicle was injected icv in freely fed C57Bl/6 mice immediately before the dark phase and food intake was monitored using an automated episodic feeding monitoring system. Nesfatin-130–59 (0.1, 0.3, 0.9nmol/mouse) induced a dose-related reduction of 4-h food intake by 28%, 49% and 49% respectively resulting in a 23% decreased cumulative 24-h food intake compared to vehicle at the 0.3nmol/mouse dose (p<0.05). The peak reduction occurred during the 3rd (-96%) and 4th hour (-91%) post injection and was associated with a reduced meal frequency (0–4h: −47%) and prolonged inter-meal intervals (3.1-times) compared to vehicle (p<0.05), whereas meal size was not altered. In contrast, neither nesfatin-11–29 nor nesfatin-160–82 reduced dark phase food intake at equimolar doses although nesfatin-160–82 prolonged inter-meal intervals (1.7-times, p<0.05). Nesfatin-130–59 is the active core of nesfatin-11–82 to induce satiety indicated by a reduced meal number during the first 4h post injection. The delayed onset may be indicative of time required to modulate other hypothalamic and medullary networks regulating nocturnal feeding as established for nesfatin-1.

Phase 1 Study of a Recombinant Mutant Protective Antigen of Bacillus anthracis

A phase 1 study of a recombinant mutant protective antigen (rPA) vaccine was conducted in 186 healthy adults aged 18 to 45 years. Volunteers were randomized to receive one of three formulations of rPA (formalin treated, alum adsorbed, or both), in 10- or 20-μg dosages each, or the licensed vaccine, AVA. Three injections were given at 2-month intervals and a 4th 1 year after the 3rd. Vaccinees were examined at the clinic once following each injection, at 48 to 72 h postinjection. Adverse reactions were recorded in diaries for 7 days. Sera were collected before each injection and 1 week after the 1st, 2 weeks after the 3rd and 4th, and 1 year after the 4th. Serum anti-PA IgG was assayed by enzyme-linked immunosorbent assay (ELISA) and toxin neutralization assay (TNA). All formulations at both dosages were safe and immunogenic, inducing booster responses, with the highest antibody levels following the 4th injection (354 to 732 μg/ml). The lowest levels were induced by the formalin-only-treated rPA; there was no statistical difference between levels induced by alum-adsorbed and formalin-treated/alum-adsorbed rPA or by the two dosages. The antibody levels declined in all groups during the 1-year intervals after the 3rd and 4th injections but less so during the 2nd year, after the 4th injection (fold decreases were 10 to 25 versus 3.4 to 7.0, P < 0.001). There were too few AVA recipients for statistical comparisons, but their antibody levels followed those of rPA. Anti-rPA measured by ELISA correlated with TNA titers (r = 0.97). These data support studying alum-adsorbed rPA in children.

Fetal Human Liver Progenitor Cells: A Potential Immune-Privileged Cellular Source from Non Heart Beating Donors

Fetal Human Liver Progenitor Cells: A Potential Immune-Privileged Cellular Source from Non Heart Beating Donors

English

Rational drug usage, clinical regulations and pharmaceutical care are important features of health practice.&nbsp;Thus; we have aimed case report to elaborate the clinical pharmaceutical services. A 3 year old girl, was presented in a hospital with minor burn that happened by spilt hot water on her wrist. She was in severe pain. The emergency ward staff applied an ointment. The doctor&nbsp;instructed injection Dormicum two times with short gape and then 3rd injection of Pavulon (Pancuronium). Patient went silent and started losing consciousness and eventually passed away. In view of the above facts and circumstances the inquiry board and judiciary took&nbsp;action against the negligence, mishandling. While; going through the investigation the inquiry board did not bothered the channel through which this injection came without proper check that is therapeutical drug monitoring, prescription reviewing, bio-safety and pharmaceutical evaluation. Furthermore, it is not a single death but representation of so many casualties because of the wrong medications. The medial practice needs some more legal, professional and principled restrictions to make sure the safe and correct therapy plans. The clinical legislation, drug rules and medical practice seriously need the attention of judiciary, society and leadership to assure the safety of precious live. &nbsp; Key words:&nbsp;Wrong medication, medical malpractice, professional negligence, pharmaceutical services,&nbsp;Suo-moto notice of chief justice of high court.

Intravitreal bevacizumab (AVASTIN®) in the treatment of diabetic macular oedema

Abstract Purpose To report visual acuity and anatomic response to intravitreal bevacizumab(IVB) injection in patients with persistent diabetic macular oedma(DMO). Methods Clinic notes of 19 patients (20eyes)were retrospectively evaluated. VA in ETDRS, central retinal thickness (CRT) and number of IVB (1.25mg/0.05mi)injections required was recorded. All patients underwent at least one FFA prior to IVB therapy. Re‐treatment was PRN based on persistent or increase in fluid with drop of VA.Previous treatments included triamcinolone (IVTA),grid (micropulse and green) lasers.Follow up was every 6weeks. Results The mean age was 69.5±6.9 (range 59‐81) years. All eyes had previous lasers. Seven eyes had both laser and IVTA. Mean number of bevacizuamab injection was 1.15. Three eyes received 2nd injection at mean of 13.3weeks (range 4‐24) and 1 eye a 3rd injection at 9weeks. The average interval between the grid lasers and avastin therapy was 18.9weeks (SD=10.35). Mean follow up was 6.45±1.1 (6‐9) months. The mean baseline VA was 42.51±16.24 and mean baseline CRT was 471±95.23. The mean change in vision at 1st 3rd and 6th month was +10.8, +11.8 and +13.2(p=0.0867) The mean reduction in CRT from baseline was ‐101.36µm,‐167.7µm and‐149.25µm at month 1, 3 and 6 respectively. (p= 0.00912). Three patients completed 9months follow up with a mean change in VA of +10.83 and CRT of‐105µm. Twenty‐ five percent gained &gt;15letters, 75% maintained stable vision and none lost more than 15 letters. No complications recorded. Conclusion Intravitreal bevacizumab 1.25mg was followed by improvement of vision and a significant reduction in persistent DMO. Further, larger studies are needed to evaluate the long term efficacy and safety of IVB on persistent diabetic macular oedema.

Central melanocortin stimulation increases phosphorylated perilipin A and hormone-sensitive lipase in adipose tissues

Norepinephrine (NE) released from the sympathetic nerves innervating white adipose tissue (WAT) is the principal initiator of lipolysis in mammals. Central WAT sympathetic outflow neurons express melanocortin 4-receptor (MC4-R) mRNA. Single central injection of melanotan II (MTII; MC3/4-R agonist) nonuniformly increases WAT NE turnover (NETO), increases interscapular brown adipose tissue (IBAT) NETO, and increases the circulating lipolytic products glycerol and free fatty acid. The WAT pads that contributed to this lipolysis were inferred from the increases in NETO. Because phosphorylation of perilipin A (p-perilipin A) and hormone-sensitive lipase are necessary for NE-triggered lipolysis, we tested whether MTII would increase these intracellular markers of lipolysis. Male Siberian hamsters received a single 3rd ventricular injection of MTII or saline. Trunk blood was collected at 0.5, 1.0, and 2.0 h postinjection from excised inguinal, retroperitoneal, and epididymal WAT (IWAT, RWAT, and EWAT, respectively) and IBAT pads. MTII increased circulating glycerol concentrations at 0.5 and 1.0 h, whereas free fatty acid concentrations were increased at 1.0 and 2.0 h. Western blot analysis showed that MTII specifically increased p-perilipin A and hormone-sensitive lipase only in fat pads that previously had MTII-induced increases in NETO. Phosphorylation increased in IWAT at all time points and IBAT at 0.5 h, but not RWAT or EWAT at any time point. These results show for the first time in rodents that p-perilipin A can serve as an in vivo, fat pad-specific indictor of lipolysis and extend our previous findings showing that central melanocortin stimulation increases WAT lipolysis.

In vivo Fluorescence Imaging of Muscle Cell Regeneration by Transplanted EGFP-labeled Myoblasts

In vivo fluorescence imaging (FLI) enables monitoring fluorescent protein (FP)-labeled cells and proteins in living organisms noninvasively. Here, we examined whether this modality could reach a sufficient sensitivity to allow evaluation of the regeneration process of enhanced green fluorescent protein (eGFP)-labeled muscle precursors (myoblasts). Using a basic FLI station, we were able to detect clear fluorescence signals generated by 40,000 labeled cells injected into a tibialis anterior (TA) muscle of mouse. We observed that the signal declined to approximately 25% on the 48 hours of cell injection followed by a recovery starting at the second day and reached a peak of approximately 45% of the original signal by the 7th day, suggesting that the survived population underwent a limited run of proliferation before differentiation. To assess whether transplanted myoblasts could form satellite cells, we injured the transplanted muscles repeatedly with cardiotoxin. We observed a recovery of fluorescence signal following a disappearance of the signal after each cardiotoxin injection. Histology results showed donor-derived cells located underneath basal membrane and expressing Pax7, confirming that the regeneration observed by imaging was indeed mediated by donor-derived satellite cells. Our results show that FLI is a powerful tool that can extend our ability to unveil complicated biological processes such as stem cell-mediated regeneration.

Development of pentylenetetrazole kindling in rats is associated with an increase in hippocampal doublecortin expression

Doublecortin is a 50 kDa protein normally expressed in neurons at early stages of their differentiation. In this study, we investigated the doublecortin expression in rat hippocampus during chronic pentylenetetrazole administration (kindling). Rats were decapitated after the 1st, 3rd, 5th, and 13th injection of the chemoconvulsant at a subconvulsive dose; doublecortin levels were assessed using Western-blot analysis. Antibodies to doublecortin stained two proteins with molecular weights of 50 and 70 kDa. Doublecortin expression increased after 13 injections of pentylenetetrazole, which indicates intensification of neurogenesis in the hippocampus of rats with increased seizure readiness. The 70 kDa proteins were found in both neocortex and hippocampus; changes in their expression did not correlate with doublecortin expression.

Plasma concentration of amrubicinol in gamma phase in patients treated for 3 days with amrubicin and hematological toxicities

e14606 Background: Amrubicinol (AMR-OH) is an active metabolite of amrubicin (AMR), a novel synthetic 9-amino-anthracycline derivative. The time-concentration profile of AMR-OH exhibits a continuous long plateau slope in terminal phase. To determine the relationships between the steady-state plasma concentration of AMR-OH and treatment effects and toxicities associated with AMR therapy, we performed a PK/PD study of AMR in patients with lung cancer treated with AMR alone or the combination of AMR + cisplatin (CDDP). Methods: AMR was given at a dose of 30 or 40 mg/m2 on days 1 to 3. Plasma samples were collected at pretreatment, 24 hours after the 1st injection (day 2), and 24 hours after the 3rd injection (day 4). Plasma concentrations of AMR, AMR-OH, and CDDP were determined by HPLC and AAS. Percent change in neutrophil count (dANC) and the plasma concentration of AMR-OH were evaluated using a sigmoid Emax model. Results: A total of 35 patients with a median age of 65 years (range 40–78), including 10 with small cell lung cancer (SCLC), 23 with non-small cell lung cancer (NSCLC), 1 with thymic cancer, and 1 with neuroblastoma were enrolled. A total of 107 plasma samples were available for study. Mean concentrations of AMR on day 2, AMR on day 4, AMR-OH on day 2, and AMR-OH on day 4 were 8.52ng/mL+4.63, 16.55ng/mL+11.92, 7.28 ng/mL+3.56 SD, and 13.35ng/mL+5.56 (mean ± SD), with significant increase from day 2 to day 4 for both AMR (p&lt;0.0001) and AMR-OH (p&lt;0.0001). Significant relationships were observed between AMR-OH on day 4 and the toxicity grades of leukopenia, neutropenia, and anemia (p=.018, p=.012, and p=.025). Thrombocytopenia grade exhibited a tendency toward relationship with AMR-OH on day 4 (p = .081). No relationships were observed between drug concentrations and responses. The plasma concentration of AMR-OH on day 4 was positively correlated with dANC in the group of all patients, as well as in patients treated with AMR alone and in patients with co-administration with CDDP using a sigmoid Emax model. Conclusions: The plasma concentration of AMR-OH on day 4 was correlated with hematological toxicities in patients treated with AMR. Assessment of plasma concentration of AMR-OH at one time point might enable prediction of hematological toxicities. No significant financial relationships to disclose.

Knee vs hip single-joint intra-articular hyaluronic acid injection in patients with both hip and knee osteoarthritis: a pilot study

This paper aims to compare the results of single-joint knee vs hip hyaluronic acid (HA) injections in patients with osteoarthritis (OA) involving both the knee and hip joints. Thirty-eight patients who were diagnosed to have both hip and knee OA were enrolled. Patients were divided into two groups to receive HA injection three times at 1-week intervals either to the hip or knee joints. Pain level during activities and rest was measured by using visual analog scale (VAS). Western Ontario and McMaster University Osteoarthritis Index (WOMAC 5-point Likert 3.0) was also used prior to the injections and 1 month after the 3rd injection. In the knee injection group, the intragroup analysis revealed significant improvements in VAS activity pain, VAS rest pain, and WOMAC pain values following injection when compared with preinjection values, while no significant difference was detected in WOMAC stiffness, WOMAC physical function, and WOMAC total values. In the hip injection group, VAS activity pain, VAS rest pain, WOMAC pain, WOMAC stiffness, WOMAC physical function, and WOMAC total values showed significant improvement after the injection when compared with preinjection values. Although statistically not significant (p > 0.05), the comparison of the differences (preinjection-postinjection) between the groups demonstrated higher values in the hip injection group. We imply that intra-articular single-joint HA injections either to the knee or hip joints in OA patients with involvement of both of these joints are effective with regard to pain and functional status.