Post-treatment Day Research Articles

18 Impact of in-feed vs. in-water antibiotic administrations on the fecal prevalence and antimicrobial susceptibilities of Campylobacter and Salmonella in piglets

Abstract Campylobacter and Salmonella are common food borne pathogens in the gut of pigs and are shed in the feces. The control of these bacteria in pigs is of importance in reducing the potential for transmission to humans. In swine, oral route, either in-feed or in-water, is by far the most common route of administration of antibiotics. Because the distribution of the antibiotic in the gut and the dosage are different, the impact of in-feed vs. in-water administration of antibiotics on the fecal shedding of food borne pathogens, Campylobacter and Salmonella, and on the development of antimicrobial resistance (AMR) in gut bacteria is a largely unexplored area. Therefore, a study was conducted to compare the effects of in-feed and in-water antibiotic administration on fecal prevalence of Campylobacter and Salmonella and AMR development in nursery piglets. A total of 1,296 weaned piglets were allocated into pens (48 pens; 27 pigs per pen) distributed in a single room. Pens were assigned randomly to six treatment groups; Control (No antibiotic), In-feed chlortetracycline (CTC), In-water CTC, In-feed tiamulin, In-water tiamulin, and a combination of CTC and tiamulin (In-feed). Fresh fecal samples were collected randomly from 5 of 27 piglets from each pen on pre-treatment (days -7, 0), treatment (days 7, 14) and post-treatment (days 21, 28) phases. Bacterial isolations and identifications were done by culture method and PCR, respectively. Overall prevalence of Campylobacter and Salmonella were 18.2% (262/1,440) and 3.9% (56/1,440) respectively. Speciation of Campylobacter isolates indicated C. hyointestinalis (17.9%; 258/1,440) and C. coli (0.3%; 4/1,440). Pigs from control group had a higher prevalence (P< 0.05) of both Campylobacter and Salmonella when compared to other treatment groups. Both treatment and post-treatment phases had a significant effect on the prevalence of Campylobacter and Salmonella (P< 0.05).

Exopolysaccharide Producing Bifidobacterium animalis subsp. lactis Strains Modify the Intestinal Microbiota and the Plasmatic Cytokine Levels of BALB/c Mice According to the Type of Polymer Synthesized.

Bacteria-host interactions are mediated by different microbial associated molecular patterns which are most often surface structures such as, among others, exopolysaccharides (EPSs). In this work, the capability of two isogenic EPS-producing Bifidobacterium animalis subsp. lactis strains to modulate the gut microbiota of healthy mice, was assessed. Each strain produces a different type of polymer; the ropy strain S89L synthesized a rhamnose-rich, high-molecular weight EPS in highest abundance than the non-ropy DMS10140 one. BALB/c mice were orally fed for 10 days with milk-bifidobacterial suspensions and followed afterward for 7 post-intervention days (wash-out period). The colonic content of mice was collected in several sampling points to perform a metataxonomic analysis. In addition, the influence of specific microbial clades, apparently stimulated by the ropy and non-ropy strains, on mouse plasmatic cytokine levels was investigated through hierarchical association testing. Analysis of 16S rRNA gene sequences showed that the abundance of Firmicutes phylum significantly increased 7 days after cessing the treatment with both strains. The relative abundance of Alloprevotella genus also rose, but after shorter post-treatment times (3 days for both DMS10140 and S89L strains). Some bacterial clades were specifically modulated by one or another strain. As such, the non-ropy DMS10140 strain exerted a significant influence on Intestinomonas genus, which increased after 4 post-administration days. On the other hand, feeding with the ropy strain S89L led to an increase in sequences of Faecalibaculum genus at 4 post-treatment days, while the abundance of Erysipelotrichaceae and Lactobacillaceae families increased for prolonged times. Association testing revealed that several lactobacilli and bifidobacterial significantly stimulated by ropy S89L strain were positively associated with the levels of certain cytokines, including IL-5 and IL-27. These results highlight relevant changes in mice gut microbiota produced after administration of the ropy S89L strain that were associated to a potential immune modulation effect.

Is conventional management of primary spontaneous pneumothorax appropriate?

This study aimed to evaluate the recurrence rate after primary and secondary conservative treatments and to clarify the validity of current primary spontaneous pneumothorax management by comparing secondary conservative treatment and surgical outcomes. Data from 166 patients with primary spontaneous pneumothorax treated at a single site between September 2015 and March 2019 were retrospectively evaluated. Patient characteristics of those who received primary conservative therapy (n = 166) and secondary conservative therapy (n = 28) were summarized. The outcomes from patients who experienced recurrence (n = 64) were compared based on those who underwent surgery (n = 24) and those who underwent secondary conservative therapy (n = 28). The post-treatment day 60 recurrence rate was 27.1 and 49.5% cases in the primary and secondary treatment groups, respectively, which was significantly higher after secondary treatment than after primary treatment with conservative therapy (p = 0.032). The post-treatment one-year recurrence rate was 13.5 and 57.9% in patients who underwent surgery and secondary conservative treatment, respectively; secondary conservative treatment resulted in a significantly higher recurrence rate than surgery (p < 0.001). There is evidence for guidelines that recommend surgery for recurrent primary spontaneous pneumothorax after primary conservative therapy based on its lower and more delayed post-treatment recurrence rate than secondary treatment with conservative therapy.

Identification Of Strn-Ntrk2 Rearrangement In A High Grade Sarcoma, With Good Clinical Response To Firstline Larotrectinib Therapy

Abstract Casestudy Gene fusions involving tropomyosin receptor kinase genes, NTRK (NTRK1-3), are important in tumorigenesis. Larotrectinib, a selective NTRK inhibitor, is recently approved to treat NTRK fusion positive solid tumors. We herein report a case of soft tissue sarcoma harboring two STRN-NTRK2 gene fusions, with good clinical response to firstline larotrectinib treatment. Results A 35 year-old female presented with pain in the right gluteal region, and a large solid mass without overlying erythema, edema and induration was identified. Initial MRI study showed a heterogenous, vascular and partially necrotic mass (16.5 x 12.9 x 10.4 cm) centered in the right gluteus medius and maximus muscles. A core biopsy of the mass showed a cellular mesenchymal neoplasm with round/ovoid cells, high mitosis (21 per 10 HPFs) and focal staghorn type vessels, reminiscent of solitary fibrous tumor. However, STAT6 immunostaining was negative. Additional immunostains show no specific lineage. Our in-house NGS fusion panel showed two in-frame STRN- NTRK2 fusions, containing the same 5’ partner sequence (exon 1-3) of STRN, with the 3’ fusion partner starting from either the exon 15 or the exon 16 of NTRK2. Due to the large size and location of the tumor, larotrectinib was initiated as firstline therapy. The patient noticed a quick amelioration of tumor related pain, and a significant shrinkage of the size of tumor following the initial 7-day treatment. On post-treatment day 52, MRI showed the tumor significantly decreased in size to 7.7 x 7.4 x 6.6 cm with satisfactory symptomatic relief. Conclusion NTRK2 fusions are relatively rare when compared with NTRK1 and NTRK3, especially in sarcoma. Of note, the only other report in the literature of NRTK2 fusion- positive sarcoma also showed SFT-like morphology, and the patient responded well to larotrectinib as second line therapy.

Identification Of Strn-Ntrk2 Rearrangement In A High Grade Sarcoma, With Good Clinical Response To Firstline Larotrectinib Therapy

Abstract Casestudy Gene fusions involving tropomyosin receptor kinase genes, NTRK (NTRK1-3), are important in tumorigenesis. Larotrectinib, a selective NTRK inhibitor, is recently approved to treat NTRK fusion positive solid tumors. We herein report a case of soft tissue sarcoma harboring two STRN-NTRK2 gene fusions, with good clinical response to firstline larotrectinib treatment. Results A 35 year-old female presented with pain in the right gluteal region, and a large solid mass without overlying erythema, edema and induration was identified. Initial MRI study showed a heterogenous, vascular and partially necrotic mass (16.5 x 12.9 x 10.4 cm) centered in the right gluteus medius and maximus muscles. A core biopsy of the mass showed a cellular mesenchymal neoplasm with round/ovoid cells, high mitosis (21 per 10 HPFs) and focal staghorn type vessels, reminiscent of solitary fibrous tumor. However, STAT6 immunostaining was negative. Additional immunostains show no specific lineage. Our in-house NGS fusion panel showed two in-frame STRNNTRK2 fusions, containing the same 5’ partner sequence (exon 1-3) of STRN, with the 3’ fusion partner starting from either the exon 15 or the exon 16 of NTRK2. Due to the large size and location of the tumor, larotrectinib was initiated as firstline therapy. The patient noticed a quick amelioration of tumor related pain, and a significant shrinkage of the size of tumor following the initial 7-day treatment. On post-treatment day 52, MRI showed the tumor significantly decreased in size to 7.7 x 7.4 x 6.6 cm with satisfactory symptomatic relief. Conclusion NTRK2 fusions are relatively rare when compared with NTRK1 and NTRK3, especially in sarcoma. Of note, the only other report in the literature of NRTK2 fusion- positive sarcoma also showed SFT-like morphology, and the patient responded well to larotrectinib as second line therapy.

Epidemiological and Clinical Studies in Cats with Thoracolumbar Vertebral Fracture and Luxation and Its Management

Background: Thoraco-lumbar spinal trauma frequently results in neurological dysfunction of varying degrees in domestic cats. The consequences may be permanent disability or death depending on severity and segment of spinal cord affected. Assessment of primary damage to spinal cord is important to prevent secondary damage and complications arising from neurological deficit. Since assessment of neurological dysfunction and its treatment in spinal trauma in cats is an evolving field, the present study was undertaken with an objective to record and associate epidemiological data and clinical examination findings at the time of presentation with clinical outcome on 60th post-treatment day in cats with thoracolumbar vertebral fracture and luxation, for a better clinical approach in this condition. Methods: 24 affected cats were assessed epidemiologically and clinically at the time of presentation. All the cats were treated either conservatively or surgically based on extent of neurological dysfunction and severity of injury on radiographic evaluation. Post-treatment observations was then correlated with epidemiological data and clinical findings to assess the clinical outcome. Result: In the present study, sub-adult semi-domicile cats were most affected and major cause was automobile accident. Presence of concomitant injuries (30%) affected recovery. Thoracolumbar spinal cord segment (T3-L3) was most affected (71%). Among 16 cats with unstable vertebral fractures, 5/6 cats recovered after surgical treatment and 2/10 cats recovered after conservative treatment. Survival and recovery was poor when degree vertebral canal displacement was more than 70% (10 cats). All 7 cats with grade 1 or grade 2 and one cat with grade 3 neurological dysfunction recovered after conservative treatment. 5 cats with grade 3 and 2 cats with grade 4 neurological dysfunction recovered after surgical treatment. Selection of surgical treatment provided better recovery in cats with unstable vertebral fractures and greater degree of neurological dysfunction.

Effect of injectable eprinomectin on milk quality and yield of dairy ewes naturally infected with gastrointestinal nematodes

The objective was to investigate the effect of injectable eprinomectin on milk yield and quality of dairy ewes naturally infected with gastrointestinal nematodes when grazing in communal pastures. Onehundred and fifty (150) clinically healthy adult lactating ewes, equally selected from 3 farms, were included in the study. On day -7, the ewes on each farm were randomly allocated into 2 equal groups of 25 animals (n=50): Control group (C) and Treated group (T). On day 0, ewes in group T were given a single subcutaneous injection of eprinomectin at a dose rate of 0.2 mg/kg bodyweight. Ewes in group C were left untreated during the whole experiment. Ewes in group T with a fecal egg count (FEC) >300 eggs per g on day +60 were treated again. Fecal samples were individually collected on days -7, 0, +30, +60, +90, +120 for FEC estimations and coprocultures. On days -7, 0, +30, +60 and +90, individual milk yield (MY) was recorded using ICAR approved volumetric milk meters. Energy corrected milk yield (ECMY) for 6% fat was also calculated. Moreover, individual milk samples were collected on each day for determination of chemical composition [fat (F%), protein (P%) and lactose (L%) content] and somatic cell counts (SCC). On each day, individual fat and protein yield (FY and PY, respectively) were calculated. Total lactation MY, total ECMY, total FY and total PY were computed. The most prevalent parasite at pre-treatment and post-treatment days was Haemonchus spp. The overall efficacy on days +30 and +90 was 97.27 % and 98.80 %, respectively. In two out of the three farms, 80 % and 91.3 % of T ewes received a second treatment on day +60, due to high parasitic burden. Treatment had a significant effect (P=0.033) on MY with an average benefit of 8%. No significant effects of treatment were observed on the other parameters, although values were constantly numerically higher for treated ewes compared to control ones. In this field trial, injectable eprinomectin had a high overall efficacy and a beneficial effect on daily milk yield.

Dynamic Contrast-Enhanced Magnetic Resonance Imaging as Imaging Biomarker for Vascular Normalization Effect of Infigratinib in High-FGFR-Expressing Hepatocellular Carcinoma Xenografts.

Overexpression of fibroblast growth factor receptor (FGFR) contributes to tumorigenesis, metastasis, and poor prognosis of hepatocellular carcinoma (HCC). Infigratinib-a pan-FGFR inhibitor-potently suppresses the growth of high-FGFR-expressing HCCs in part via alteration of the tumor microenvironment and vessel normalization. In this study, we aim to assess the utility of dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) as a non-invasive imaging technique to detect microenvironment changes associated with infigratinib and sorafenib treatment in high-FGFR-expressing HCC xenografts. Serial DCE-MRIs were performed on 12 nude mice bearing high-FGFR-expressing patient-derived HCC xenografts to quantify tumor microenvironment pre- (day 0) and post-treatment (days 3, 6, 9, and 15) of vehicle, sorafenib, and infigratinib. DCE-MRI data were analyzed using extended generalized kinetic model and two-compartment distributed parameter model. After treatment, immunohistochemistry stains were performed on the harvested tumors to confirm DCE-MRI findings. By treatment day 15, infigratinib induced tumor regression (70% volume reduction from baseline) while sorafenib induced relative growth arrest (185% volume increase from baseline versus 694% volume increase from baseline of control). DCE-MRI analysis revealed different changes in microcirculatory parameters upon exposure to sorafenib versus infigratinib. While sorafenib induced microenvironment changes similar to those of rapidly growing tumors, such as a decrease in blood flow (F), fractional intravascular volume (vp), and permeability surface area product (PS), infigratinib induced the exact opposite changes as early as day 3 after treatment: increase in F, vp, and PS. Our study demonstrated that DCE-MRI is a reliable non-invasive imaging technique to monitor tumor microcirculatory response to FGFR inhibition and VEGF inhibition in high-FGFR-expressing HCC xenografts. Furthermore, the microcirculatory changes from FGFR inhibition manifested early upon treatment initiation and were reliably detected by DCE-MRI, creating possibilities of combinatorial therapy for synergistic effect.

Complex toxin binder mycotox® ng influence on the hepatotoxic effect of aflatoxin B1 in experimental treated goslings

Abstarct. The aim of the present investigation was to evaluate the effects of aflatoxin B1 and Mycotox NG applied either independently or together, on blood total protein, albumin, blood glucose, total bilirubin, triglycerides, cholesterol, enzyme activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AP), gamma-glutamyl transferase (γ GT), lactate dehydrogenase (LDH) and changes in liver morphology. At the same time, the potential of supplementation of feed with a mycosorbent (Mycotox NG) was evaluated. Experiments were carried out with 40 1-day-old Toulouse geese from mixed sexes divided into one control and three treatment groups (n=10). Groups were as followed: Group I – control (0 mg/kg AFB1 not supplemented with Mycotox NG); Group II (0.5 g/kg Mycotox NG), Group III (0.5 mg/kg AFB1) and Group IV (0.5 mg/kg AFB1 and 0.5 g/kg Mycotox NG). In this study, commercially available geese of Toulouse strain were reared from day one to forty two days in the deep litter system of management and the birds were divided into four groups. Normal feed tested free of aflatoxin (AFB1), was given to the control (Group – 1). 0.5 g/kg Mycotox was supplemented with the feed to Group 2, Aflatoxin (0.5 mg/kg feed) was supplemented with the feed to Group 3 and Mycotox Ng (0.5 g/kg feed) + 0.5 mg/kg feed AFB1 was supplemented with the feed to Group 4. The duration of the experiments was 42 days. The monitored blood chemical parameters were analysed on post treatment days 21 and 42. In birds treated only with AFB1, (group III) increased blood activities of studied enzymes. At the same time, blood total protein, albumin, cholesterol, glucose and triglycerides were reduced as compared to controls. The observed histopathological changes in the liver consisted in various extent of dystrophy (congestion, vacuolar and granular dystrophy, round cell proliferation, necrobiotic changes, hyperplasia of gallbladder epithelium). The addition of mycosorbent (Mycotox NG) to the feed of Groups IV reduced substantially the changes in blood chemistry and the severity and frequency of liver histological lesions. The addition of mycosorbent (Mycotox NG) to the feed of Groups IV reduced substantially the changes in blood chemistry and the severity and frequency of liver histological lesions.

No Evidence for Ceftobiprole-Induced Immune Hemolytic Anemia in Three Phase 3 Clinical Trials.

PurposeDrug-induced immune hemolytic anemia (DIIHA) is a rare but serious adverse event associated with a number of drugs, including second- and third-generation cephalosporins. A positive direct antiglobulin test (DAT) is a reliable finding in DIIHA, but positive results without evidence of hemolysis can occur, particularly in hospitalized patients. There have been no reports of hemolytic anemia in four previous Phase 3 trials or from post-marketing surveillance of the advanced-generation, broad-spectrum cephalosporin, ceftobiprole. The aim of this analysis was to review the incidence of positive DAT results and any evidence of hemolytic anemia from three recent Phase 3 trials of ceftobiprole.Patients and MethodsPatients were enrolled in three Phase 3 randomized controlled trials: 94 pediatric patients with pneumonia received ceftobiprole in the BPR-PIP-002 trial; 335 adults with acute bacterial skin and skin structure infections received ceftobiprole in the TARGET trial; and 201 adults with Staphylococcus aureus bacteremia have been randomized 1:1 to ceftobiprole or daptomycin ± aztreonam in the ongoing ERADICATE trial. In all three trials, DAT results were obtained at baseline, and follow-up tests were performed either at the test of cure (TOC) visit (BPR-PIP-002), end-of-treatment (EOT) visit (TARGET), or both EOT and post-treatment Day 70 visits (ERADICATE).ResultsIn the BPR-PIP-002 trial, five patients (all ceftobiprole treated) had a documented negative DAT result at baseline followed by a positive result at the TOC visit. One patient in the ongoing, blinded ERADICATE trial had a positive DAT result at both baseline and EOT. Results from other laboratory investigations showed no evidence of hemolytic anemia in these patients. No positive DAT results were reported in the TARGET trial.ConclusionNo evidence of hemolytic anemia associated with ceftobiprole was observed in either adults or children across several indications in this analysis of three large Phase 3 trials.

Acacia farnesiana pods (plant: Fabaceae) possesses anti-parasitic compounds against Haemonchus contortus in female lambs

Acacia farnesiana pods are rich in secondary metabolites and their biological activities have been recorded as antibacterial, antioxidant and anthelmintic. Previously, an in vitro bioguided study showed the important ovicidal and larvicidal effects of an organic fraction (EtOAc-F) from a hydroalcoholic extract of A. farnesiana pods against Haemonchus contortus. The present study aimed to assess the in vivo anthelmintic effect of EtOAc-F from A. farnesiana pods on the H. contortus faecal egg elimination in female lambs and on the infective larvae (L3) population reduction in coprocultures. The EtOAc-F was obtained from a hydroalcoholic extract from A. farnesiana pods through chromatographic procedures; additionally, some secondary compounds were identified using high-performance liquid chromatography (HPLC). Twenty-one ‘Katahdin’ crossbred female lambs ranging from three to four months of age, with body weights 21.9 ± 0.39 kg were used. Animals were orally infected with H. contortus (L3) by a single dose of 350 L3/kg BW. Three experimental groups (n = 7) were assigned as follows: 1) Control (untreated), 2) Albendazole, as a positive control (at 7.5 mg/kg BW, unique dose) and 3) EtOAc-F (at 100 mg/kg BW, once every third day, with three applications in total). Individual faecal samples were collected once a week for 5 weeks (at days 38, 45, 52, 59 and 66) post-treatment, to measure the faecal egg counts (FEC) and to obtain the H. contortus (L3) population from faecal cultures. The highest FEC reduction caused by EtOAc-F was 67.7%; meanwhile, albendazole showed a total FEC reduction after the second week post-treatment (day 45). On the other hand, the fraction caused an important reduction in the larval population in coprocultures (54.3–68.5%). The phytochemical analysis revealed the presence of galloyl derivatives and flavonoids as major compounds. The A. farnesiana pods could serve as a natural anthelmintic for the control of H. contortus, and perhaps for controlling other parasites of veterinary importance.

Decreased hemoglobin after initial treatment is associated with treatment resistance in Kawasaki disease in Kobayashi risk stratification.

Although Kawasaki disease (KD) is known to cause anemia, the relationship between its prognosis and hemoglobin level is less well known. We herein performed a secondary analysis of data from a previous study (Post-RAISE). Children aged 6months to 2years at the time of KD diagnosis were enrolled. Decreased hemoglobin was defined as a decrease > 10% on post-treatment day 2 compared with the level prior to the initial therapy. Of 1253 KD cases, treatment resistance was observed in 209 (17%) and was significantly more frequent in the decreased hemoglobin group (37/167 vs. 172/1086, P = 0.041). Multivariable logistic regression analysis revealed that decreased hemoglobin was associated with resistance to the initial treatment (odds ratio 1.53; 95% confidence interval 1.00-2.33; P = 0.048). Decreased hemoglobin was significantly associated with resistance to initial treatment in patients with KD.

Efficacy of Sub-Tenon Micro-Perfusion of Cyclophosphamide in Rabbits with Severe Ocular Inflammation.

PurposeTo explore the feasibility of cyclophosphamide (CP) via a sub-Tenon micro-perfusion system (SMS) in rabbits, and assess its therapeutic efficacy in severe ocular inflammation.Materials and MethodsDistribution and pharmacokinetics of CP were evaluated in vivo, and the concentrations of CP in plasma, vitreous humor, and retina/choroid were quantitated by ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) at different time points. After induction of severe experimental uveitis, rabbits were divided into three groups (n=8 in each): the SMS group, subconjunctival injection (SI) group, and control group. Clinical inflammatory score was assessed in rabbits. Electroretinography and histopathology were performed on post-treatment day 8. Statistical analyses were performed using Mann–Whitney and Kruskal–Wallis tests. P-value less than 0.05 was considered significant.ResultsThe concentrations of CP in vitreous humor and retina/choroid in the SMS group were significantly higher than that of the SI group at 3, 6, 10, and 24 hours (P<0.01), while plasmatic CP concentrations were comparable at all time points in the SMS group and SI group (P>0.05). The SMS group showed significantly less inflammation compared to the control group and SI group. Furthermore, the restoration of retinal structure and function were more obvious in the SMS group compared with conventional SI application.ConclusionSub-Tenon micro-perfusion of CP exhibited satisfied therapeutic efficacy in rabbits with severe ocular inflammation and may provide a promising alternative for controlling ocular inflammatory disease and immune-mediated ocular diseases.

Comparison of therapeutic efficacy of different drugs against Trypanosoma vivax on experimentally infected cattle

In this study, we evaluated the therapeutic efficacy of diminazene diaceturate at a dose of 7 mg/kg (DA), imidocarb dipropionate at 4.8 mg/kg (IMD), isometamidium chloride at 0.5 and 1.0 mg/kg (ISM 0.5 and ISM 1.0) and combinations applied through different methods to treat Trypanosoma vivax in experimentally infected calves. Thirty male Girolando calves were kept indoors and infected intravenously with T. vivax trypomastigotes (approximately 1 × 106). On D-1, the calves were randomized based on the quantity of infecting parasites per animal, yielding six groups of five animals each: G1: positive control group without treatment; G2 animals treated with DA on Day 0 intramuscularly (IM); G3 animals treated with IMD on Day 0 and D + 14 subcutaneously; G4 animals treated with ISM 0.5 on Day 0 IM; G5 animals treated with ISM 1.0 on Day 0 IM; G6 animals received DA on Day 0 and ISM 1.0 on D + 14, both IM. Throughout 180 days, blood samples were collected for the evaluation of T. vivax using the Woo, Brener and PCR methods. The results indicated that the treatment protocols with DA and/or ISM 0.5 and ISM 1.0 had high efficacy (100 %) against T. vivax. Interestingly, cattle that received ISM remained free of parasites until D + 180. In contrast, animals treated with IMD had relapsed T. vivax detected on the 10th and 14th days post-treatment (DPT). Cattle that received ISM 1.0 did not exhibit relapsed T. vivax in the blood, even after reinfection performed on the 50th DPT. However, treatment with DA on Day 0 failed to prevent a new infection of T. vivax on the 50th DPT. The animals that received ISM 1.0 had a transient decrease in packed cell volume similar to that found in the control group. The reappearance of T. vivax in herds in Brazil treated with DA likely occurred due to the short half-life of the drug and not necessarily due to T. vivax resistance to DA.

Effect of activating chimeric receptor on IL-15 armored NK cell on providing in vitro and in vivo antigen specific tumor response.

e15016 Background: Colorectal cancer liver metastases (CRCLM) are a major source of morbidity and mortality. Historically, curative therapy has been limited to surgical resection, but only a small fraction of patients are eligible. Cellular immunotherapy has shown promise in hematologic cancers, but challenges related to solid tumor therapy remain with optimal cell trafficking, elevated interstitial fluid pressures (IFP), and immunosuppression. We hypothesized that engineered natural killer (NK) cells expressing a natural killer group 2, member D (NKG2D) activating chimeric receptor (ACR) and membrane bound IL-15 (NKX101) would increase anti-tumor activity in vitro and in vivo utilizing our established regional delivery strategies. Methods: In vitro cytotoxicity and cytokine release of NKX101 cells or non-transduced NK cells (NT-NK) derived from the same donor were determined by co-culture systems with HCT116 cells that endogenously express NKG2D ligands. CRCLM-bearing NSG™ mice were treated with NKX101, NT-NK, or vehicle (CTRL) via portal vein (PV) for regional delivery (RD) or tail vein (TV) for systemic delivery (SD). Tumor burden (TB) was measured via tumor bioluminescence (TBL) and histopathology (HP). Flow cytometry (FC) determined the quantity of cells delivered. Student’s t-test and Mann-Whitney tests were performed for statistical comparisons. Results: NKX101 transduction efficiencies ranged between 63.5 – 75.6% across 3 separate healthy donors. EC50 values derived from a 4-hour cytotoxicity assay for NKX101 vs. NT-NK were 3-4 fold lower with the greatest difference observed at the 1:1 effector-to-target (E:T) ratio (mean percent cytotoxicity: 72% vs. 20%, p = 0.001). In vitro cytokine assessment revealed 2.0-2.6 fold increases in IFN-γ, GM-CSF, and TNF-α levels compared to NT-NK cells (p &lt; 0.0001 across all groups). In vivo, FC showed 2.89-fold increase in cell delivery using RD vs. SD on PTD1 (n = 3, p = 0.006). TBL was improved with 5 x 106 cells via PV vs. TV (n = 6) from post-treatment day (PTD) 1-7, with greatest difference seen on PTD7 (12.9 vs. 42.6, p = 0.07). HP analysis showed reduction of TB at PTD7 with PV treatment. Conclusions: NKX101 demonstrated improvements in in vitro cytotoxicity and pro-inflammatory cytokine release. RD techniques in vivo revealed increased cell delivery and improved tumor control. Further studies are underway to confirm our initial findings and understand NKX101 cellular kinetics and susceptibility to immunosuppression in the liver, along with planned clinical evaluation in Phase 1 trials.

Temsirolimus combined with etoposide and cyclophosphamide for relapsed/refractory acute lymphoblastic leukemia: Therapeutic advances in Childhood Leukemia Consortium (TACL 2014-001) trial.

10512 Background: PI3K/mTOR signaling, a critical pathway in cell proliferation, metabolism, and apoptosis, is often dysregulated in acute lymphoblastic leukemia (ALL). A phase 1 trial of the mTOR inhibitor temsirolimus combined with etoposide and cyclophosphamide was performed in children with relapsed/refractory (r/r) ALL. Methods: Temsirolimus was administered intravenously (IV) on days 1 and 8 with cyclophosphamide 440 mg/m2 and etoposide 100 mg/m2 IV daily days 1-5. The starting dose level (DL) of temsirolimus was 7.5 mg/m2 (DL1) with escalation to 10 mg/m2 (DL2), 15 mg/m2 (DL3), and 25 mg/m2 (DL4). MRD was performed centrally. PI3K pathway inhibition was measured by phosphoflow cytometry (PFC) analysis of peripheral blood (PB) from treated patients (pts). Results: Sixteen heavily pretreated r/r ALL pts ages 2-19 years with marrow blasts &gt; 25% were enrolled; 15 were evaluable [10 B-ALL/5 T-ALL]. One dose-limiting toxicity (DLT) of grade (Gr) 4 pleural and pericardial effusions with pneumonitis/lung infection leading to Gr 5 cardiorespiratory arrest occurred in a pt treated at DL3. No further DLTs were seen in the DL3 expansion and DL4 cohorts. Gr 3/4 non-hematologic toxicities occurring in ≥ 3 pts included febrile neutropenia, elevated ALT, hypokalemia, mucositis, and tumor lysis syndrome and were independent of dose. Of 15 evaluable pts, 4 (27%; 2 B-ALL/2 T-ALL) had a complete response (CR) after cycle 1, comprised of 1 pt at each DL. Three had MRD &lt; 0.01%. Three pts (20%; 2 B-ALL/1 T-ALL) had partial response (PR). Overall response rate (CR+PR = ORR) was 47%. Pharmacodynamic PFC studies compared phosphoprotein levels pre (day 0) and post treatment (days 3-5) in 9 consenting pts with available PB. All tested pts showed basal activation of PI3K pathway signaling. Dose-dependent inhibition of mTOR targets phosphorylated (p) S6 and/or p4EBP1 was observed in 9/9 and 6/9 pts, respectively, following temsirolimus and chemotherapy treatment. Various patterns of compensatory upregulation of pPI3K, pmTOR, pAkt, and/or pERK was observed. Conclusions: Temsirolimus at 25 mg/m2 combined with salvage etoposide and cyclophosphamide has an acceptable safety profile in high-risk pediatric patients with r/r ALL. Responses were observed at all DLs. mTOR target inhibition was achieved and appeared to correlate with dose level. Future testing of other PI3K/mTOR pathway inhibitors in combination with chemotherapy may be warranted with a goal of further increasing response in r/r ALL. Clinical trial information: NCT01614197.

Time-Dependent Memory and Gait Improvement by Intranasally-Administered Extracellular Vesicles in Parkinson's Disease Model Rats.

We have recently demonstrated that extracellular vesicles (EVs) derived from the human teeth stem cells improve motor symptoms and normalize tyrosine hydroxylase (TH) expression in the nigrostriatal structures of Parkinson's disease (PD) model rats obtained by 6-hydroxydopamine (6-OHDA) unilateral injection into the medial forebrain bundle (MFB). The aim of this study was to clarify: (1) how long therapeutic effects persist after discontinuation of 17-day intranasal administration of EVs in 6-OHDA rats; (2) may EVs reverse cognitive (learning/memory) dysfunction in these PD model rats; (3) whether and how the behavioral improvement may be related to the expression of TH and Nissl bodies count in the nigrostriatal structures. Our results demonstrated that in 6-OHDA rats, gait was normalized even ten days after discontinuation of EVs administration. EVs successfully reversed 6-OHDA-induced impairment in spatial learning/memory performance; however, the beneficial effect was shorter (up to post-treatment day 6) than that revealed for gait improvement. The shorter effect of EVs coincided with both full normalization of TH expression and Nissl bodies count in the nigrostriatal structures, while slight but significant increase in the 6-OHDA-decreased Nissl count persisted in the substantia nigra even on the post-treatment day 20, supposedly due to the continuation of protein synthesis in the living cells. The obtained data indicate the usefulness of further studies to find the optimal administration regimen which could be translated into clinical trials on PD patients, as well as to clarify other-apart from dopaminergic-neuromodulatory pathways involved in the EVs mechanism of action.

Improved trafficking and tumor control with regional, pressure-enabled drug delivery of anti-CEA CAR-T cells for colorectal liver metastases in a murine model

Abstract Introduction Efficacy of cellular immunotherapy for liver metastases (LM) has been limited by poor trafficking and elevated intra-tumoral pressures. In a phase 1b study, we found that regional, pressure-enabled drug delivery (PEDD, TriSalus Life Sciences, Inc.) improved LM CAR-T delivery. We hypothesized that PEDD of CAR-T in a murine model can improve LM control. Methods LM bearing CD45.2 mice were treated with CD45.1 anti-CEA CAR-T via systemic delivery (SD, tail vein) or regional delivery (RD, portal vein) at low pressure (LPRD) vs. high pressure (HPRD) to model PEDD. Flow rates were calibrated to achieve LPRD or HPRD. Tumor bioluminescence (TB), flow cytometry (FC), immunofluorescence (IF), &amp; quantitative polymerase chain reaction (qPCR) were assessed on various post-treatment days (PTD). Results FC of liver non-parenchymal cells on PTD7 showed increased CAR-T density in HPRD &amp; LPRD (9.4% &amp; 11.0%, p = 0.03 &amp; 0.002) vs. SD (3.7%). qPCR of the CAR transgene supported this observation, but was not significant (HPRD 135.8, p = 0.76, LPRD 108.6, p = 0.40 vs. SD 69.8). TB was lower in RD vs. SD up to PTD7 (log2 fold-change HPRD −1.3 &amp; LPRD −0.9 vs. SD 4.3, p = 0.005 &amp; 0.006). Tumor proliferation by Ki67 IF was significantly reduced with RD vs. control (HPRD &amp; LPRD, p &amp;lt; 0.001 &amp; p = 0.05) with HPRD superior to LPRD (p = 0.03). Serum liver enzymes were not significantly elevated using HPRD vs. LPRD at PTD7 (AST/ALT 154.7/36.3 vs. 139.1/29.1, p = 0.84/0.67), but did rise at PTD17, correlating with tumor progression (AST/ALT 800.7/229.3 vs. 387.5/139.5, p = 0.10/0.28). Conclusions RD increased LM CAR-T delivery and HPRD showed evidence of improved CAR-T trafficking and tumor control relative to LPRD. Further clinical testing of PEDD is planned for LM cellular therapy.

Observation on the effect of liquid paraffin as adjuvant therapy in treating residual wounds of patients with severe burns

Objective: To observe the effect of liquid paraffin as adjuvant therapy in treating residual wounds of patients with severe burns. Methods: From January 2016 to October 2017, 58 patients with residual wounds in the later stage of severe burns were admitted to Beijing Jishuitan Hospital. Those patients met the inclusion criteria of this study and were enrolled in this retrospective cohort study. All the patients received wound disinfection, petrolatum gauze dressing change and silver ion-bearing antimicrobial dressing every other day to treat the residual wound until wound healing. From January to November 2016, 29 patients who were treated with iodophor and normal saline to clean and disinfect the wound before dressing change were enrolled in normal saline group, including 22 males and 7 females, aged (38±12) years. From December 2016 to October 2017, 29 patients who were treated with iodophor and liquid paraffin to clean and disinfect the wound before dressing change were enrolled in liquid paraffin group, including 20 males and 9 females, aged (37±12) years. The effective rate of treatment, infection control rate, and bacterial positive ratio of wound on post first treatment day (PFTD) 7, 14, and 21 and the complete wound healing time of patients were compared between the two groups. Data were statistically analyzed with independent sample t test, Wilcoxon rank-sum test, chi-square test, and Pearson chi-square test, continuity correction chi-square test, and Fisher's exact probability test after Bonferroni correction. Results: (1) On PFTD 7, 14, and 21, the effective rates of wound treatment in patients of liquid paraffin group were 34% (10/29), 59%(17/29), and 79%(23/29), respectively, which were significantly higher than 10% (3/29), 24% (7/29), and 38% (11/29) of normal saline group (χ(2)=5.857, 7.108, 10.235, P<0.05 or P<0.01). (2) The wound infection control rates in patients of liquid paraffin group on PFTD 7 and 14 were 79% (23/29) and 90% (26/29), respectively, which were significantly higher than 31% (9/29) and 52% (15/29) of normal saline group (χ(2)=13.663, 8.321, P<0.01). The wound infection control rate in patients of liquid paraffin group on PFTD 21 was 100% (29/29), which was significantly higher than 69% (20/29) of normal saline group (P<0.01). (3) The bacterial positive ratios in the wounds of liquid paraffin group were similar to those of normal saline group on PFTD 7 and 14 (χ(2)=2.097, 2.583, P>0.05). On PFTD 21, the wound bacterial positive ratio of liquid paraffin group was 8% (3/39), which was significantly lower than 49% (17/35) of normal saline group (χ(2)=13.625, P<0.01). (4) The complete wound healing time of patients in liquid paraffin group was 15.0 (11.0, 20.0) d, which was significantly shorter than 22.0 (15.0, 27.5) d of normal saline group (Z=2.256, P<0.05). Conclusions: In the treatment of residual wounds of severe burn patients, liquid paraffin as adjuvant therapy can effectively improve the treatment effective rate and the infection control rate of wounds, decrease the bacterial positive ratio, and shorten the wound healing time.

Interactive tissue reactions of 1064‐nm focused picosecond‐domain laser and dermal cohesive polydensified matrix hyaluronic acid treatment in in vivo rat skin

Picosecond-domain laser treatment using a microlens array (MLA) or a diffractive optical element (DOE) generates micro-injury zones in the epidermis and upper dermis. To investigate interactive tissue reactions between MLA-type picosecond laser pulses and cohesive polydensified matrix hyaluronic acid (CPMHA) filler in the dermis. In vivo rats with or without CPMHA pretreatment were treated with a 1064-nm picosecond-domain neodymium:yttrium-aluminum-garnet (Nd:YAG) laser using an MLA or DOE. Skin samples were obtained at post-treatment days 1, 10, and 21 and histologically and immunohistochemically analyzed. Picosecond-domain Nd:YAG laser treatment with an MLA-type or a DOE-type handpiece generated fractionated zones of pseudo-cystic cavitation along the lower epidermis and/or upper papillary dermis at Day 1. At Day 21, epidermal thickness, dermal fibroblasts, and collagen fibers had increased. Compared to CPMHA-untreated rats, rats pretreated with CPMHA showed marked increases in fibroblasts and collagen fibers in the papillary dermis. Immunohistochemical staining for the hyaluronic acid receptor CD44 revealed that MLA-type picosecond laser treatment upregulated CD44 expression in the basilar epidermis and dermal fibroblasts. We suggest that the hyaluronic acid-rich environment associated with CPMHA treatment may enhance MLA-type picosecond-domain laser-induced tissue reactions in the epidermis and upper dermis.