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Adenosine triphosphate binding cassette proteins (ATP-ABC) transporters belong to a broad superfamily of proteins in all living organisms that have essential physiological functions. They are necessary for transporting a wide range of substrates across lipid membranes. ABC transport proteins share two conserved domain architectures: the multiple transmembrane helices (TMD) nucleotide binding protein domain (NBD). Fungi have evolved active transport pathways through endogenous and exogenous toxicants. Drug efflux from ABC transporters lowers the concentration of intracellular drugs, in many pathogens and cancer cells; therefore, efflux is subjected to extensive studies in pathogens. ABC transporter gene of aspergillus versicolor as reference and evaluated metadata of ABC transporter gene in comparison from dermatophyte and non-dermatophyte fungi using fungi DB and NCBI databases. ABC transporters gene was studied using gene homology via phylogenetic analysis. Study evaluated ABC transporters sub families, number of transcripts, exon, intron numbers sizes and position using gene sequence analysis by NCBI. The evolutionary history of ABC gene analyzed by using 12 proteins of both dermatophytes. Study also explored the single nucleotide polymorphism (SNPs) homology in dermatophytes using the NCBI SNP tool. ABC protein examined five sub-families which include ABC-B, ABC-C, ABC-D, ABC-F. Phylogenetic studies on ABC transporter genes showed its emergence in three major groups of dermatophytes and non- dermatophyte. Exon number of ABC transporter gene varies from 2 to 18 and one transcript number in dermatophytes and non-dermatophytes. The evolutionary history of ABC-B was analyzed by using 12 proteins of both dermatophytes and non-dermatophytes. ABC genes in dermatophytes were found in five subfamilies ABC-B, ABC-C, ABC-D, ABC-F and ABC-G. However, ABC transporter gene family survey suggested that ABC- C family is a highly emerged sub family in dermatophytes and non-dermatophytes. Study revealed that ABC protein has been found in five sub-families of ABC transporter gene in dermatophytes. It has one transcript number in dermatophytes and non-dermatophytes. Whereas, the exon numbers are varied from 2 to 18. Moreover, ABC transporter gene phylogenetic family survey suggested that ABC- C family is highly emerged in both fungal types.

Background:Squalene epoxidase gene is an azole antifungal target enzyme in the ergosterol biosynthesis pathway in fungi—the inhibition of enzyme aggregate squalene, a type of ergosterol derivative depletion that leads to fungal growth inhibition. Squalene epoxidase widely diverges in three fungal groups of dermatophytes Trichophyton, Microsporum and Epidermophyton. Methodology:The study has demonstrated a variety of squalene epoxidase genes in the dermatophyte genome. Squalene epoxidase gene was also studied for possible single nucleotide polymorphism (SNPs) in the Trichophyton group's homologs and found multiple SNP positions that induce non-synonymous mutation and change the sequence of amino acid into protein structure that can change phenylalanine to leucine. Results:Squalene epoxidase is widely present in dermatophytes. Mutation in the squalene epoxidase gene can mislead normal sterol formation in the fungal cell membrane that induces fungal resistance against several drugs, including azoles and polyenes. Squalene epoxidase gene carries 2 - 3 transcripts and 2 - 3 number of exons. Squalene epoxidase gene has FAD-dependent oxidoreductases and NADP binding domain conserved domains in fungal groups of dermatophytes. Conclusion:This study analyzed the abundance of the squalene epoxidase gene, suggesting that squalene epoxidase gene-resistant mutants can occur naturally. Thus squalene epoxidase gene should be extensively studied in order to increase the potential of available antifungals. Link:http://aeirc-edu.com/ojs14/index.php/IJEHSR/article/view/574/651

Objectives: To compare the difference in responses regarding lapses in professionalism related to academic integrity between first and final year MBBS students, and in relation to gender. Methodology: A quantitative cross sectional study was conducted in Allama Iqbal Medical College between first and final year MBBS students over a period of one year from March 2016 to Feb 2017. A total 200 students were included 100 from each class in the study. A validated and customized 47 items Dundee poly professionalism inventory-I questionnaire was given to all students. Results: The response rate was 100%, all the students completed the Performa and returned back. Eighty percent of the first year students were between 18-20 years. Eighty percent of the final year students were 23-24 years. Out of 200 students 58.5% were females and 41.5% were males. The difference in responses regarding lapses in professionalism related to academic integrity was not statistically significant class-wise, but significant gender-wise. Conclusions: The study highlighted that there is no difference in responses regarding lapses in professionalism related to academic integrity among class wise. However significant difference was observed gender wise, more lapses were observed in Males as compare to Females.

Irrigation with sewage-contaminated water poses a serious threat to food security, particularly in developing countries. Heavy metal tolerant bacteria are sustainable alternatives for the removal of wastewater contaminants. In the present study, four lead (Pb)-tolerant strains viz. Bacillus megaterium (N8), Bacillus safensis (N11), Bacillus sp. (N18), and Bacillus megaterium (N29) were inoculated in spinach and grown in sewage water treated earthen pots separately and in combination with canal water. Results showed that Pb-tolerant strains significantly improved plant growth and antioxidant activities in spinach and reduces metal concentration in roots and leaves of spinach plants irrigated with treated wastewater. Strain Bacillus sp. (N18) followed by B. safensis (N11) caused the maximum increase in shoot length, root length, shoot fresh weight, root fresh weight, shoot dry weight, root dry weight, and leaf area compared to the uninoculated control of sewage water treated plants. These strains also improved antioxidant enzymatic activity including catalase, guaiacol peroxidase dismutase, superoxide dismutase, and peroxidases activities compared to the uninoculated control under sewage water conditions. Strain Bacillus sp. (N18) followed by B. safensis (N11) showed the highest reduction in nickel, cadmium, chromium, and Pb contents in roots and leaves of spinach compared to the uninoculated control plants treated with the sewage water. Such potential Pb-tolerant Bacillus strains could be recommended for the growth promotion of spinach after extensive evaluation under field conditions contaminated with wastewater.

Mineral nutrition of crop plants is one of the major challenges faced by modern agriculture, particularly in arid and semi-arid regions. In alkaline calcareous soils, the availability of phosphorus and zinc is critically less due to their fixation and precipitation as complexes. Farmers use fertilizers to fulfill crop requirements, but their efficacy is less, which increases production costs. Plant growth-promoting rhizobacteria (PGPR) can improve the availability of crop nutrients through solubilizing the insoluble compounds of phosphorus and zinc in soil. In the present study, a total of 40 rhizobacterial isolates were isolated from cotton rhizosphere and screened for improving cotton growth through the solubilization of phosphorus and zinc. Out of these 40 isolates, seven isolates (IA2, IA3, IA6, IA7, IA8, IA13, and IA14) efficiently solubilized insoluble rock phosphate while seven isolates (IA10, IA16, IA20, IA23, IA24, IA28, and IA30) were more efficient in solubilizing insoluble zinc oxide. In liquid media, strain IA7 (2.75 μg/mL) solubilized the highest amount of phosphate while the highest concentration of soluble zinc was observed in the broth inoculated with strain IA20 (3.94 μg/mL). Seven phosphate-solubilizing and seven zinc-solubilizing strains were evaluated using jar trial to improve the growth of cotton seedlings, and the results were quite promising. All the inoculated treatments showed improvement in growth parameters in comparison with control. Best results were shown by the combined application of IA6 and IA16, followed by the combination of strains IA7 and IA20. Based on the jar trial, the selected isolates were further characterized by plant growth-promoting characters such as siderophores production, HCN production, ammonia production, and exopolysaccharides production. These strains were identified through 16S rRNA sequencing as Bacillus subtilis IA6 (accession # MN005922), Paenibacillus polymyxa IA7 (accession # MN005923), Bacillus sp. IA16 (accession # MN005924), and Bacillus aryabhattai IA20 (accession # MN005925). It is hence concluded that the integrated use of phosphate-solubilizing and zinc-solubilizing strains as potential inoculants can be a promising approach for improving cotton growth under semi-arid conditions.

SERine Proteinase INhibitors Clade B: SERPINS, a largely dispersed family of protease inhibitors that exhibit a conformational alteration to inhibit target enzymes. Atypical serpin proteins often form aggregates that can accumulate to toxic levels within cells. SerpinB4 is a chymotrypsin-like serine proteinase inhibitor. Phylogenetic analysis of Serpin Clade B revealed no duplication throughout the mammalian clades. Thus, elucidating retention of a single copy of human serpin members in nature, endorsing its essentiality as an inhibitor. Epidemiological studies specify that a rising number of patients are inclined to develop chronic liver disease due to SerpinB4 overexpression. This upregulation is therefore not in fine fettle for the human physiology. Molecular docking protocol classified compound 57, a novel inhibitory compound, as the ligand, best able to fit the binding pocket with a Gold Score of 94. A two hundred nanosecond molecular dynamics simulation of docked SerpinB4 illustrated a steady binding pattern of ligand in the protein's active site. Pursuing this, trajectory analysis was implemented to assess various characteristics of the docked system in terms of function of time. These findings may act effectively in designing effective remedies against SerpinB4.

Background: Adenosquamous carcinoma of the pancreas (ASCP) is a very rare and highly aggressive variant of pancreatic ductal adenocarcinoma (PDA), accounting for 0.5-4% of all pancreatic cancer cases in the US. Current data indicates that epigenetic changes and Myc overexpression lead to squamous transdifferentiation of pancreatic tumor cells and development of ASCP. Minnelide is an oral anti-super-enhancer drug that inhibits Myc expression in preclinical models of ASCP. Oral Minnelide has been previously tested in the clinic where a Phase I study determined a recommended phase 2 dose of 2 mg/day. Major toxicities included neutropenia and sepsis. We hypothesize that Minnelide will be safe and effective in patients with advanced ASCP. Methods: This Phase II open label, single arm study being conducted at the NIH Clinical Center in Bethesda, MD (NCT04896073) will enroll up to 25 evaluable participants with advanced ASCP that has been previously treated. Eligibility criteria include: histologically confirmed ASCP (≥30% malignant squamous component), presence of metastatic, recurrent, or locally advanced unresectable disease, and progression or intolerance ≥1 prior systemic treatment for advanced disease. All participants will self-administer Minnelide at 2mg PO daily on days 1-21 of each 28-day cycle, for up to 12 cycles in the absence of disease progression or unacceptable toxicity. Restaging scans will be performed every 8 weeks. Optional biopsies will be requested at baseline, mid-cycle 1 and at time of progression. The primary objective is to determine the antitumor activity of Minnelide by evaluating the disease control rate. Secondary objectives are determination of drug safety in this patient population, progression free survival, and overall survival. Exploratory objectives include assessment of Minnelide effect on tumor cell epigenetics, Myc expression and immune cell populations using on-treatment tumor biopsy tissues. Changes in circulating tumor DNA and circulating immune cell populations upon treatment will also be evaluated. A Simon optimal two-stage phase II trial design will be used. If 3 of the first 12 patients experience disease control, then enrollment will continue up to 25 evaluable participants. If ≥8 of 25 participants experience disease control this would rule out an unacceptably low clinical benefit of 20% in favor of 40% (α = 0.1, β = 0.2). This trial opened for accrual in July 2021. Citation Format: Nebojsa Skorupan, Mehwish I. Ahmad, Seth M. Steinberg, Jane B. Trepel, Derek Cridebring, Haiyong Han, Daniel D. Von Hoff, Christine Alewine. A phase II trial of the super-enhancer inhibitor Minnelide in advanced refractory adenosquamous carcinoma of the pancreas (ASCP) [abstract]. In: Proceedings of the AACR Virtual Special Conference on Pancreatic Cancer; 2021 Sep 29-30. Philadelphia (PA): AACR; Cancer Res 2021;81(22 Suppl):Abstract nr PO-054.

Purpose: LMB-100 is a recombinant immunotoxin (iTox) consisting of a mesothelin-binding Fab for targeting and a modified Pseudomonas exotoxin A payload. Preclinical studies showed that combining taxanes with iTox results in synergistic antitumor activity. The objectives of this phase I/II study were to determine the MTD of LMB-100 when administered with nanoalbumin bound (nab)-paclitaxel to patients with previously treated advanced pancreatic adenocarcinoma and to assess the objective response rate. Patients and Methods: Patients (n = 20) received fixed-dose nab-paclitaxel (125 mg/m2 on days 1 and 8) with LMB-100 (65 or 100 μg/kg on days 1, 3, and 5) in 21-day cycles for 1–3 cycles. Results: Fourteen patients were treated on the dose escalation and an additional six in the phase II expansion. MTD of 65 μg/kg was established for the combination. Dose-limiting toxicity resulting from capillary leak syndrome (CLS) was seen in two of five patients treated at 100 μg/kg and one of six evaluable phase I patients receiving the MTD. Severity of CLS was associated with increases in apoptotic circulating endothelial cells. LMB-100 exposure was unaffected by anti-LMB-100 antibody formation in five of 13 patients during cycle 2. Seven of 17 evaluable patients experienced >50% decrease in CA 19-9, including three with previous exposure to nab-paclitaxel. One patient developed an objective partial response. Patients with biomarker responses had higher tumor mesothelin expression. Conclusions: Although clinical activity was observed, the combination was not well tolerated and alternative drug combinations with LMB-100 will be pursued.

Objective: The main idea of this research work is to assess the role of magnetic resonance imaging to examine the degree of disease of spinal TB (Tuberculosis). Methodology: This research work was an elaborate descriptive carried out in the surgical department & orthopedics department in General Hospital, Lahore from August 2017 to March 2019. Sixty patients suffering from the spinal TB were the part of his research work who got admission through emergency department of the same hospital. Fifty five percent (n: 33) patients were males & 45.0% (n: 27) were female’s patients. The range of the age of patients was from 14 to 36 years. The average age of the patients was thirty-three years. We noted the clinical aspects of all the patients & we collected the elaborated information of the patients on a well-organized Performa. Confirm patients of spinal TB were the part of this research work. Patients diagnosed with no tuberculosis were not the part of this research work. Results: The very frequent symptom was backache in 63.0% (n: 38) patients. We found the compression of the spinal cord in 26.60% (n: 16) patients. Very frequent affected level was the thoracolumbar spine present in 45.0%. We found the diffuse contribution as very rare & only 5.0% (n: 3) patients were available with the involvement of more than one level. The most basic modality of imaging was the plain radiography. The most important effective investigation was MRI. The findings of the MRI for spinal TB were decreased intervertebral space between discs in 95.0% patients, wedge body collapse in 30.0% (n: 18) patients, whole body destruction in 20.0% (n: 12) & para-spinal abscess in 40.0% (n: 24) patients. We found the calcification in 30.0% (n: 18) patients & cord compression in 26.60% (n: 16) patients. Conclusion: The most effective and valuable investigation in the patients suffering from spinal TB is MRI. It provides the data about the disease extend & it is the best method to provide guidelines for the surgical intervention. The use of the MRI is necessary during the follow up to check the response of the whole treatment. Key Words: Destruction, methodology, compression, MRI, spinal, TB, suffering, response, treatment, therapy, discs.

Objectives: This research study aimed to find out the relationship between smoking of cigarettes and the pulmonary TB. Methodology: This transverse research study conducted in Jinnah Hospital, in Lahore city located South of Punjab and in the middle of the country Pakistan. We reviewed the medical charts of the patients from TB between 2016 to the year of 2018. Total 61 patients having age from 15 to 96 years with confirm pulmonary TB were the part of case group & one hundred and twenty two persons with matched age & gender with no TB (patients getting treatment in other wards of hospital) were the part of the control group. We collected the information about the smoking condition, amount of the cigarettes smoked in a day and smoking duration from the medical charts of the participants. SPSS. 22 was in use for the comparison of the collected information of both groups. Chi-square test was in use for the comparison of the rate of the smoking of cigarettes in both groups. Results: Out of total sixty one patients, 68.90% (n: 42) patients were smoker, whereas out of total one hundred and twenty two controls, 18.0% (n: 22) persons were smoker. The proposed OR (Odds Ratio) of the association between habit of cigarette smoking & TB was 10.10 with CI of 95%. The average pocket-year of smoked cigarette (Twenty per pocket) in patients & controls persons were 15.90 ± 13.70 & 13.50 ± 9.10 correspondingly. Conclusion: The results of this research study concluded that there is strong association between pulmonary TB and habit of cigarette smoking. Therefore, smoking of cigarette is a positive risk factor for the acquisition of the tuberculosis. Key Words: Pulmonary, tuberculosis, smoking, cigarette, TB, methodology, acquisition, SPSS, comparison, controls.&nbsp

3051 Background: LMB-100 recombinant immunotoxin consists of a mesothelin-binding Fab for targeting a modified Pseudomonas exotoxin A payload to tumors. Previous clinical trials demonstrated that almost all patients formed anti-drug-antibodies (ADAs) to LMB-100 that made administration beyond cycle 2 ineffective. Tofacitinib is an oral JAK inhibitor that prevented formation of ADAs against a closely related immunotoxin in pre-clinical studies. The primary objective of the dose escalation cohort was assessment of safety and tolerability of LMB-100 given with tofacitinib to patients with mesothelin-expressing solid tumors. The primary objective of the expansion cohort was to determine whether co-administration of tofacitinib delays formation of neutralizing LMB-100 ADAs. Methods: Patients (n = 13) with pancreatic adenocarcinoma and other mesothelin-expressing solid tumors (n = 3; cholangiocarcinoma, appendix, cystadenocarcinoma) were treated for up to 3 cycles with LMB-100 as a 30-minute infusion on days 4, 6, and 8 at two dose levels (100 and 140 mcg/kg) and co-treated with oral tofacitinib for the first 10 days of the cycle (10 mg BID). Results: Dose level 1 of LMB-100 was started at 100 mcg/kg one dose level below the single agent MTD. Dose escalation to 140 mcg/kg (dose level 2) resulted in DLTs in 2 of the 3 patients treated: grade 3 cardiac toxicity and grade 4 hyponatremia, both attributed to capillary leak syndrome. Ultimately, 7 patients were treated at dose level 1 without DLTs and 100 mcg/kg was chosen as the LMB-100 dose for the expansion cohort. The last of 6 patients treated in the expansion cohort developed grade 4 pericardial effusion leading to early closure of the study for toxicity. No objective responses were seen. Of the 8 patients who received two cycles of treatment at MTD, 4 met prespecified criteria for ADA prevention, and 2 patients who went on to receive cycle 3 had detectable LMB-100 plasma drug levels after administration. Conclusions: LMB-100 was unable to be co-administered safely with tofacitinib. ADA formation was prevented in 2 patients through 3 cycles, a rare occurrence. Clinical trial information: NCT04034238.

TPS452 Background: LMB-100 recombinant immunotoxin consists of a mesothelin-binding Fab for targeting with a modified Pseudomonas exotoxin A payload. Formation of anti-drug antibodies (ADAs) is thought to contribute significantly to limited clinical efficacy of LMB-100 seen in prior clinical trials. Most patients develop clinically meaningful ADAs after 1-2 cycles of LMB-100, resulting in rapid neutralization of LMB-100 during subsequent cycles and undetectable plasma drug levels. Tofacitinib is an oral Janus Kinase-1 and -3 (JAK) inhibitor approved by the FDA for the treatment of rheumatoid arthritis and ulcerative colitis. Pre-clinical studies have shown that tofacitinib can prevent the formation of ADAs against recombinant immunotoxin (Onda et al. Journal of Immunology 2014), and that co-administration of tofacitinib with LMB-100 increases immunotoxin serum half- life in mice and anti-tumor efficacy (Simon et al. JCI Insight 2019). We hypothesize that co-administration of tofacitinib with LMB-100 will prevent or delay the formation of high titer ADAs to LMB-100, such that 2 effective cycles of immunotoxin can be administered to patients. Methods: This phase I clinical trial consists of a dose escalation phase using 3+3 design to determine the maximum tolerated dose (MTD) of LMB-100 that can be administered with tofacitinib in participants (n = 18 max) with mesothelin-expressing solid tumors, followed by a dose expansion phase at the MTD for participants (n = 15) with pancreatic adenocarcinoma or extrahepatic cholangiocarcinoma. The primary objective of the expansion phase is to determine whether co-administration of tofacitinib delays formation of neutralizing anti-LMB-100 ADAs for cycle 2 of treatment as measured by LMB-100 plasma drug levels. A positive outcome will be reached if percent of participants achieving threshold LMB-100 drug levels during cycle 2 increases from 50% to 80% (83.6% probability if >10 of 15 evaluable participants meet this milestone). Plasma drug levels during cycle 3 will also be analyzed as a secondary endpoint. Key inclusion criteria include adults with histologically confirmed previously treated solid tumor malignancies. Participants will receive tofacitinib 10 mg twice daily on days 1-10 and LMB-100 at 65, 100, or 140 mcg/kg on days 4, 6 and 8 of a 21-day cycle. The dose escalation phase has been completed and enrollment onto dose expansion phase is ongoing. Clinical trial information: NCT04034238.

3553 Background: LMB-100 recombinant immunotoxin consists of a mesothelin-binding Fab for targeting a modified Pseudomonas exotoxin A payload. Previous Phase 1 clinical testing of a 30-minute LMB-100 “short” infusion format identified a serum half-life of ~1 hour. Pre-clinical data suggested that extending infusion time could improve anti-tumor efficacy by increasing tumor cell duration of exposure to LMB-100. The primary objective of this study was to determine the safety and tolerability of administering LMB-100 in a long infusion format over 24-48 hours alone or with nab-paclitaxel chemotherapy in patients with mesothelin-expressing solid tumors. Methods: Patients (n = 15) with pancreatic adenocarcinoma and other mesothelin-expressing solid tumors (n = 3; mesothelioma, colon, and ampullary cancers) treated on 3 dose levels received long infusion of LMB-100 (65 or 100 mcg/kg/day) for 24 hour on Days 1 and 4 (n = 6) or 48 hour on Day 1 (n = 9) with or without a loading dose (40 mcg/kg over 30 minutes) for up to 2 cycles. In the second arm, patients (n = 5) with pancreatic adenocarcinoma were treated with LMB-100 over 24 hours on Day 1 concurrently with nab-paclitaxel (125 mg/m2) for up to 3 cycles. Results: DLT of proteinuria (grade 3) in one patient and acute kidney injury (grade 1) in one patient were observed amongst patients receiving 100 mcg/kg/day over 48 hours and 24 hours, respectively. No objective responses were seen but all patients receiving nab-paclitaxel had > 50% decrease in CA 19-9. Patients at all single agent dose levels (8 of 10 evaluable) developed high titer anti-drug antibodies (ADAs) against LMB-100. Those with ADAs (8 of 8) had undetectable cycle 2 peak plasma LMB-100 concentration. Development of high titer ADAs occurred more frequently with long infusion than seen previously with “short” infusion LMB-100. Most long infusion patients (19 of 20) developed increased serum IL-6 within 24 hours of LMB-100 infusion. However, the systemic inflammatory response to LMB-100 (as measured by increased serum CRP) which occurs in most “short” infusion patients was not observed. Conclusions: Long infusion format LMB-100 is generally well tolerated but immunogenicity limits treatment to 1 effective cycle. No anti-tumor efficacy of the single agent was observed. Clinical trial information: NCT02810418 .

Due to continuous growth of world population, there is dire need of serious efforts and innovative approaches to meet food demands through sustainable production practices, improvement in supply chain, and control of food wastage. All these efforts should ensure the access to nutritious food to all suffering from hunger and malnutrition. Due to intensive crop cultivation and use of synthetic fertilizers, soil health is seriously deteriorating. However, soil fertility can be improved by incorporating legumes in the cropping system and/or use of rhizobial inoculants, which not only increase nitrogen fixation but also improve soil fertility and crop production through several other attributes such as phosphate solubilization, siderophores production, phytohormones production, enzymes synthesis, and exopolysaccharides production. Moreover, these bacteria can be helpful for improvement in crop production on marginal lands due to their tolerance against various biotic and abiotic stresses. All these characteristics make rhizobia equally important for non-legumes as for legumes. The use of rhizobial inoculants can ensure improvement in crop productivity and environment sustainability by enhancing soil fertility and reduction in use of synthetic chemical fertilizers. Present review focuses on important plant growth-promoting mechanisms of rhizobia and the use of these rhizobia for sustainable crop production through improvement in crop nutrition, physiology, productivity, and stress tolerance of crop plants. The potential of the synergistic use of rhizobia with other soil microorganisms for sustainable agriculture has also been elucidated with examples, followed by their future prospects.

BackgroundLow phosphorus availability limits crop production in alkaline calcareous soils in semi-arid regions including Pakistan. Phosphate solubilizing bacteria may improve crop growth on alkaline calcareous soils due to their ability to enhance P availability.MethodsTwenty rhizobacterial isolates (Q1–Q20) were isolated from rhizosphere of cotton and characterized for their growth promoting attributesin vitro. The selected phosphate solubilizing isolates were further screened for their ability to improve cotton growth under axenic conditions (jar trial). The phosphorus solubilization capacities of selected strains were quantified and these strains were identified through 16S rDNA sequencing.ResultsIsolates Q2, Q3, Q6, Q7, Q8, Q13 and Q14 were able to solubilize phosphate from insoluble sources. Most of these isolates also possessed other traits including catalase activity and ammonia production. The growth promotion assay showed that Q3 was significantly better than most of the other isolates followed by Q6. Maximum root colonization (4.34 × 106cfu g−1) was observed in case of isolate Q6 followed by Q3. The phosphorus solubilization capacities of these strains were quantified, showing a maximum phosphorus solubilization by Q3 (optical density 2.605 ± 0.06) followed by the Q6 strain. The strain Q3 was identified asBacillus subtilis(accession #KX788864) and Q6 asPaenibacillussp. (accession #KX788865) through 16S rDNA sequencing.DiscussionThe bacterial isolates varied in their abilities for different growth promoting traits. The selected PGPRBacillus subtilisstrain Q3 andPaenibacillussp. strain Q6 have multifarious growth promoting traits including ability to grow at higher EC and pH levels, and phosphorus solubilizing ability. These strains can efficiently colonize cotton roots under salt affected soils and help plants in phosphorus nutrition. It is concluded that both strains are potential candidates for promoting cotton growth under alkaline conditions, however further investigation is required to determine their potential for field application.

Class B G-protein coupled receptors are involved in a wide variety of diseases and are a major focus in drug design. Migraines are a common problem, and one of their major causative agents is the class B G-protein coupled receptor, Calcitonin gene-related peptide (CGRP) receptor, a target for competitive drug discovery. The calcitonin receptor-like receptor generates complexes with a receptor activity-modifying protein, which determines the type of receptor protein formed. The CGRP receptor comprises a complex formed from the calcitonin receptor-like receptor and receptor activity-modifying protein 1. In this study, an in silico docking approach was used to target the calcitonin receptor-like receptor in the bound form with receptor activity-modifying protein 1 (CGRP receptor), as well as in the unbound form. In both cases, the resulting inhibitors bound to the same cavity of the calcitonin receptor-like receptor. The twelve evaluated compounds were competitive inhibitors and showed efficient inhibitory activity against the CGRP receptor and Calcitonin receptor-like receptor. The two studied quinoline derivatives demonstrated potentially ideal inhibitory activity in terms of binding interactions and low range nano-molar inhibition constants. These compounds could prove helpful in designing drugs for the effective treatment of migraines. We propose that quinoline derivatives possess inhibitory activity by disturbing CGRP binding in the trigeminovascular system and may be considered for further preclinical appraisal for the treatment of migraines.

Purpose: LMB-100 recombinant immunotoxin (iTox) consists of a mesothelin-binding Fab for targeting and a modified Pseudomonas exotoxin A payload. Preclinical studies showed that combining taxanes with iTox results in synergistic antitumor activity. The primary objectives of this phase I/II study were to determine the maximum tolerated dose (MTD) of LMB-100 when given with nanoalbumin bound (nab)-paclitaxel to participants with previously treated advanced pancreatic adenocarcinoma and to assess the objective response rate. Patients and Methods: Patients (n = 20) received fixed-dose nab-paclitaxel (125 mg/m2 on Days 1 and 8) with LMB-100 (65 or 100 mcg/kg on Days 1, 3, and 5) in 21-day cycles. Treatment was limited to 2-3 cycles by protocol. Safety and tolerability, pharmacokinetics, immunogenicity, and relationship of activity to tumor mesothelin expression were assessed. Results: Dose-limiting toxicity (DLT) of Grade 3 myalgia was observed in 2 of 5 patients treated with 100 mcg/kg. DLT of Grade 3 edema from capillary leak syndrome (CLS) was observed in 1 of 6 patients at MTD of 65 mcg/kg. Severe toxicities of Grade 4 left ventricular dysfunction and hypotension from CLS were observed in 1 patient in the phase II expansion, resulting in study closure. LMB-100 anti-drug antibody formation limited LMB-100 exposure in 5 of 13 patients during cycle 2. Seven of 17 patients with evaluable CA19-9 experienced >50% decrease of this tumor marker, and one patient developed a partial response. Response was associated with higher tumor mesothelin expression and Cycle 2 LMB-100 exposure. Conclusions: The combination caused serious toxicity. Despite some observed clinical activity, further testing of this combination will not be pursued. This abstract is also being presented as Poster B01. Citation Format: Christine C. Alewine, Mehwish Ahmad, Cody J. Peer, Zishuo Hu, Min-jung Lee, Akiro Yuno, Jessica Kindrick, Anish Thomas, Seth Steinberg, Jane B. Trepel, William D. Figg, Raffit Hassan, Ira Pastan. Phase I/II study of mesothelin-targeted immunotoxin LMB-100 with nab-paclitaxel for patients with advanced pancreatic adenocarcinoma [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2019 Sept 6-9; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2019;79(24 Suppl):Abstract nr PR07.

Purpose: LMB-100 recombinant immunotoxin consists of a mesothelin-binding Fab for targeting and a modified Pseudomonas exotoxin A payload. Previous Phase 1 clinical testing of a 30-minute LMB-100 infusion format identified dose-limiting toxicity (DLT) of capillary leak syndrome (CLS) and a serum half-life of ~60 minutes. Pre-clinical data suggested that extending infusion time could improve anti-tumor efficacy by increasing tumor cell duration of exposure to LMB-100. The primary objective of this study was to determine the safety and tolerability of administering LMB-100 over a longer infusion time. We hypothesized that reduced CMAX would result in decreased severity of off-target toxicities like CLS without decreasing anti-tumor activity. Patients and Methods: Patients (n = 15) with pancreatic adenocarcinoma and other mesothelin-expressing solid tumors (n = 3; mesothelioma, colon adenocarcinoma, ampullary carcinoma) treated on 3 dose levels received long infusion (24-48 hours) LMB-100 (65 or 100 mcg/kg/day) with or without a loading dose (40 mcg/kg over 30 minutes) for up to 2 cycles. Safety and tolerability, pharmacokinetics, immunogenicity and anti-tumor activity were assessed. Results: DLT of proteinuria (grade 3) was observed in 1 of 6 patients receiving 100 mcg/kg/day over 48 hours with pre-load. Three of 6 patients at this dose level developed significant CLS (>5 kg weight gain from fluid edema) that was not dose-limiting. No CLS was observed in the 6 patients who received 100 mcg/kg over 24 hours on days 1 and 4 with pre-load. Increased immunogenicity was observed with the long infusion compared to standard administration format: 8 of 9 patients receiving 2nd cycle treatment had undetectable plasma LMB-100 concentration during this cycle that was associated with the development of high titer anti-LMB-100 antibodies. Progressive disease was observed in 11 of 12 evaluable patients, and none of 8 evaluable patients with pancreatic adenocarcinoma had a clinically significant decline in CA 19-9 tumor marker. Conclusions: Long infusion format of delivery for LMB-100 can reduce CLS but increases immunogenicity. No anti-tumor activity was observed. Citation Format: Christine Alewine, Mehwish Iqra Ahmad, Cody J Peer, Seth Steinberg, William D Figg, Raffit Hassan, Ira Pastan. Phase I study of mesothelin-targeted immunotoxin LMB-100 given as a long infusion [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A081. doi:10.1158/1535-7163.TARG-19-A081

307 Background: LMB-100 is a Pseudomonas exotoxin A-based immunotoxin that targets mesothelin (MSLN). MSLN is expressed by >75% of pancreatic adenocarcinomas (PDAC). LMB-100 kills MSLN-expressing cells by irreversibly modifying elongation factor-2 to halt protein synthesis. Phase I studies of LMB-100 defined the maximum tolerated dose (MTD) of 140 mcg/kg IV given on D1, 3 and 5 of a 21-day cycle. Development of anti-drug antibodies (ADAs) limited patient drug exposure beyond cycle 2. Our pre-clinical data showed that combination of LMB-100 with a taxane resulted in synergistic anti-tumor activity. Methods: We conducted a phase I single center study (standard 3+3 design) to determine the MTD of LMB-100 given with nab-paclitaxel in patients with previously treated advanced PDAC. LMB-100 was given on D1, 3 and 5 of a 21-day cycle, and nab-paclitaxel (125 mg/m2) on D1 and D8. Initial patients could receive a maximum of 4 cycles, but subsequently a 2-cycle maximum was employed. Results: Fourteen patients (median age 58) were enrolled. Two of 6 patients experienced DLTs at the 100 mcg/kg dose of LMB-100 (myalgia- 2 pts, fatigue- 1 pt, hypotension- 1 pt; all grade 3). One of 8 patients had DLT at the 65 mcg/kg dose (edema, urine output decrease- both grade 3). Other toxicities related to LMB-100 included hypoalbuminemia, edema-associated weight gain, hyponatremia, fatigue, drug fever, infusion-related reaction, hypophosphatemia, nausea and anorexia. One patient died on treatment from complications of bowel perforation attributed to cancer. All patients achieved detectable serum levels of LMB-100 during the first cycle, even those with pre-existing ADAs, and 5 of 8 did so during cycle 2. One patient receiving the 65 mcg/kg dose had a confirmed partial response, and CA 19-9 dropped by > 50% in 5 of 8 evaluable patients. Conclusions: MTD of LMB-100 is 65 mcg/kg given with nab-paclitaxel on this schedule. Anti-tumor activity was observed. A phase II cohort is currently being accrued. Clinical trial information: NCT02810418.

Nanoparticles (NPs) that are ∼100 nm in diameter can potentially cause toxicity in the central nervous system (CNS). Although NPs exhibit positive aspects, these molecules primarily exert negative or harmful effects. Thus, the beneficial and harmful effects should be compared. The prevalence of neurodegenerative diseases, such as Alzheimer disease, Parkinson disease, and some brain tumors, has increased. However, the major cause of these diseases remains unknown. NPs have been considered as one of the major potential causes of these diseases, penetrating the human body via different pathways. This review summarizes various pathways for NP-induced neurotoxicity, suggesting the development of strategies for nanoneuroprotection using in silico and biological methods. Studies of oxidative stress associated with gene expression analyses provide efficient information for understanding neuroinflammation and neurodegeneration associated with NPs. The brain is a sensitive and fragile organ, and evolution has developed mechanisms to protect it from injury; however, this protection also hinders the methods used for therapeutic purposes. Thus, brain and CNS-related diseases that are the cause of disability and disorder are the most difficult to treat. There are many obstacles to drug delivery in the CNS, such as the blood brain barrier and blood tumor barrier. Considering these barriers, we have reviewed the strategies available to map NPs using biological techniques. The surface adsorption energy of NPs is the basic force driving NP gathering, protein corona formation, and many other interactions of NPs within biological systems. These interactions can be described using an approach named the biological surface adsorption index. A quantitative structural activity relationship study helps to understand different protein-protein or protein-ligand interactions. Moreover, equilibrium between cerebrovascular permeability is required when a drug is transferred via the circulatory system for the therapy of neurodegenerative diseases. Various drug delivery approaches, such as chemical drug delivery and carrier-mediated drug delivery, have been established to avoid different barriers inhibiting CNS penetration by therapeutic substances. Developing an improved understanding of drug receptors and the sites of drug action, together with advances in medicinal chemistry, will make it possible to design drugs with greatly enhanced activity and selectivity; this may result in a significant increase in the therapeutic index.