Anti-infective Drugs Research Articles

Individual Peptides Extraction and Characteristics of Cow Colostrum Trypsin Hydrolysate with Biological Activity Assessment

The research concerned the trypsin peptides analysis of colostrum hydrolyzate. The authors isolated seven individual peptides: TT1, TT2, TT3, T(1), T1.1, T1.2, mpT. They determined the molecular weight of the peptides and the amino acid sequence in the peptide chain by mass spectrometry; peptides – using the international database Protein NCBI. The mpT peptide contains the largest number of amino acids – 49. The T1.2 peptide is similar to the membrane-active peptide BAMA Bos taurus, which opens prospects for its use as a part of antibacterial agents or as a BAS carrier. In colostrum hydrolyzate the T1 peptide has the T highest concentration (2.04 mg/ml). It consists of 17 amino acids and matches the peptide NCI_CGAP_Brn23 cDNA Homo sapiens clone, which is similar to TR: O35085 ARX HOMEOPROTEIN. The homeobox protein containing the development gene of a PRD-like class is related to the T(1) peptide. This gene in humans is located on the X chromosome and is involved in the central nervous system and pancreas development. According to the results, the researchers isolated peptides from colostrum hydrolyzate, and identified some of them. Still a man understudies the biological role of most of the determined peptides, and has not proved its safety. In this regard, the study aimed at assessing the hydrolyzate cytotoxicity using the Neutral Red Assay method on the HEK293 cell line during a 24-hour incubation. Cytotoxicity in the first, second and third measurements at a concentration of 0.1 mg/ml was: −3.3; −1.0 and 0.20; at a concentration of 0.3 mg/ml: −1.3; −3.5 and −8.7; at 0.5 mg/ml: −2.6; −0.6 and 0.4. The results indicate no toxicity. The authors studied the antimicrobial and antifungal hydrolyzate activity. Colostrum hydrolyzate demonstrates antimicrobial activity against E. coli and B. subtilis bacteria and antifungal activity against C. albicans. The results assert that the peptides are of clinical importance and can serve as a template for the anti-infective drug development.

A Turn-On–Type Fluorescence Resonance Energy Transfer Eco-friendly Method for Nitazoxanide Quantification in Pharmaceutical Dosage Form and Spiked Plasma: Evaluation of Greenness Profile Using Different Assessment Tools

A brand-new class of anti-infective drugs that work against bacteria, viruses, and protozoan parasites is nitazoxanide and related thiazolides. Thiazolides have also been shown to cause cell cycle arrest and apoptotic cell death in cancer cells in recent years. In this study, an eco-friendly, spectrofluorimetric technique that is verified, easy, and sensitive has been proposed for quantifying nitazoxanide (NTZ), a broad-spectrum antiparasitic drug. When NTZ is reduced with zinc (Zn) powder in an acidic media, a highly fluorescent product is produced. To get the highest sensitivity, different experimental conditions impacting the response were examined and optimized. Following excitation at 299 nm, scanning of the fluorescent product was done at 440 nm. The intensity of the fluorescence was proportional to the drug concentration in the range of 0.1–0.6 μg/mL. The approach was validated according to International Conference on Harmonization (ICH) guidelines, and the outcome was satisfactory. The detection and quantitation limits were calculated to be 0.013 and 0.038 μg/mL, respectively. The suggested technique was successful in analyzing commercially available NTZ dosage forms. Furthermore, the proposed technique was used to assess NTZ levels in human plasma and it was bio-analytically validated according to European Medicines Agency (EMA) guidelines. The suggested method can be used in quality control laboratories as well as in pharmacokinetic studies. In order to picture the green profile of the developed method, four greenness assessment tools have been applied. National Environmental Methods Index (NEMI), analytical Eco-Scale Assessment (ESA), Green Analytical Procedure Index (GAPI) and Analytical Greenness metric (AGREE) are the relatively most widely used metrics. So, they were utilized to perform a detailed greenness comparison between the proposed method and some of the reported methods for the determination of NTZ. The developed method was found to be an excellent green method with the highest AGREE score.

Targeted Protein Degradation for Infectious Diseases: from Basic Biology to Drug Discovery.

Targeted protein degradation (TPD) is emerging as one of the most innovative strategies to tackle infectious diseases. Particularly, proteolysis-targeting chimera (PROTAC)-mediated protein degradation may offer several benefits over classical anti-infective small-molecule drugs. Because of their peculiar and catalytic mechanism of action, anti-infective PROTACs might be advantageous in terms of efficacy, toxicity, and selectivity. Importantly, PROTACs may also overcome the emergence of antimicrobial resistance. Furthermore, anti-infective PROTACs might have the potential to (i) modulate "undruggable" targets, (ii) "recycle" inhibitors from classical drug discovery approaches, and (iii) open new scenarios for combination therapies. Here, we try to address these points by discussing selected case studies of antiviral PROTACs and the first-in-class antibacterial PROTACs. Finally, we discuss how the field of PROTAC-mediated TPD might be exploited in parasitic diseases. Since no antiparasitic PROTAC has been reported yet, we also describe the parasite proteasome system. While in its infancy and with many challenges ahead, we hope that PROTAC-mediated protein degradation for infectious diseases may lead to the development of next-generation anti-infective drugs.

Biological and Medical Aspects Related to South American Rattlesnake Crotalus durissus (Linnaeus, 1758): A View from Colombia.

In Colombia, South America, there is a subspecies of the South American rattlesnake Crotalus durissus, C. d. cumanensis, a snake of the Viperidae family, whose presence has been reduced due to the destruction of its habitat. It is an enigmatic snake from the group of pit vipers, venomous, with large articulated front fangs, special designs on its body, and a characteristic rattle on its tail. Unlike in Brazil, the occurrence of human envenomation by C. durisus in Colombia is very rare and contributes to less than 1% of envenomation caused by snakes. Its venom is a complex cocktail of proteins with different biological effects, which evolved with the purpose of paralyzing the prey, killing it, and starting its digestive process, as well as having defense functions. When its venom is injected into humans as the result of a bite, the victim presents with both local tissue damage and with systemic involvement, including a diverse degree of neurotoxic, myotoxic, nephrotoxic, and coagulopathic effects, among others. Its biological effects are being studied for use in human health, including the possible development of analgesic, muscle relaxant, anti-inflammatory, immunosuppressive, anti-infection, and antineoplastic drugs. Several groups of researchers in Brazil are very active in their contributions in this regard. In this work, a review is made of the most relevant biological and medical aspects related to the South American rattlesnake and of what may be of importance for a better understanding of the snake C. d. cumanensis, present in Colombia and Venezuela.

Over 40 Years of Fosmidomycin Drug Research: A Comprehensive Review and Future Opportunities.

To address the continued rise of multi-drug-resistant microorganisms, the development of novel drugs with new modes of action is urgently required. While humans biosynthesize the essential isoprenoid precursors isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) via the established mevalonate pathway, pathogenic protozoa and certain pathogenic eubacteria use the less well-known methylerythritol phosphate pathway for this purpose. Important pathogens using the MEP pathway are, for example, Plasmodium falciparum, Mycobacterium tuberculosis, Pseudomonas aeruginosa and Escherichia coli. The enzymes of that pathway are targets for antiinfective drugs that are exempt from target-related toxicity. 2C-Methyl-D-erythritol 4-phosphate (MEP), the second enzyme of the non-mevalonate pathway, has been established as the molecular target of fosmidomycin, an antibiotic that has so far failed to be approved as an anti-infective drug. This review describes the development and anti-infective properties of a wide range of fosmidomycin derivatives synthesized over the last four decades. Here we discuss the DXR inhibitor pharmacophore, which comprises a metal-binding group, a phosphate or phosphonate moiety and a connecting linker. Furthermore, non-fosmidomycin-based DXRi, bisubstrate inhibitors and several prodrug concepts are described. A comprehensive structure-activity relationship (SAR) of nearly all inhibitor types is presented and some novel opportunities for further drug development of DXR inhibitors are discussed.

A Novel and Efficient Platform for Discovering Noncanonical Quorum-Quenching Proteins.

Quorum sensing (QS) is a well-known chemical signaling system responsible for intercellular communication that is widespread in bacteria. Acyl-homoserine lactone (AHL) is the most-studied QS signal. Previously, bacterially encoded AHL-degrading enzymes were considered to be canonical quorum-quenching proteins that have been widely used to control pathogenic infections. Here, we report a novel platform that enabled the efficient discovery of noncanonical AHL quorum-quenching proteins. This platform initially asked bacteriologists to carry out comparative genomic analyses between phylogenetically related AHL-producing and non-AHL-producing members to identify genes that are conservatively shared by non-AHL-producing members but absent in AHL-producing species. These candidate genes were then introduced into recombinant AHL-producing E. coli to screen for target proteins with the ability to block AHL production. Via this platform, we found that non-AHL-producing Lysobacter containing numerous environmentally ubiquitous members encoded a conserved glycosyltransferase-like protein Le4759, which was experimentally shown to be a noncanonical AHL-quenching protein. Le4759 could not directly degrade exogenous AHL but rather recognized and altered the activities of multiple AHL synthases through protein-protein interactions. This versatile capability enabled Le4759 to block specific AHL synthase such as CarI from Pectobacterium carotovorum to reduce its protein abundance to suppress AHL synthesis, thereby impairing bacterial infection. Thus, this study provided bacteriologists with a unique platform to discover noncanonical quorum-quenching proteins that could be developed as promising next-generation drug candidates to overcome emerging bacterial antibiotic resistance. IMPORTANCE Targeting and blocking bacterial quorum sensing (QS), the process known as quorum quenching (QQ) is an effective mean to control bacterial infection and overcome the emerging antibiotic resistance. Previously, diverse QS signal-degradation enzymes are identified as canonical QQ proteins. Here, we provided a novel and universal platform that enabled to discover previously unidentified noncanonical QQ proteins that were unable to degrade acyl-homoserine lactone (AHL) but could block AHL generation by recognizing multiple AHL synthases via direct protein-protein interactions. Our findings are believed to trigger broad interest for bacteriologists to identify potentially widely distributed noncanonical QQ proteins that have great potential for developing next-generation anti-infectious drugs.

Application of initial anti-infectives and trends in children with septic shock

Objective: To investigate the pathogen composition, initial anti-infectives and pathogen coverage, and trends over the last 5 years in children with septic shock in pediatric intensive care unit (PICU). Methods: The single-center retrospective study included 257 children with septic shock who were admitted to PICU of Beijing Children's Hospital, Capital Medical University from 2017 to 2021. The causitive pathogen composition, initial use of anti-infective drugs, pathogen coverage, and changes in recent years were analyzed. The children were divided into sufficient and insufficient coverage groups according to whether the pathogen were sufficiently covered by initial anti-infectives; community-and hospital-acquired groups; and with and without underlying disease groups. T test, rank-sum test and Chi-square test were used for comparison between the groups to investigate the differences in pathogen, treatment and prognosis. Results: A total of 257 septic shock children were included, with 162 males and 95 females, aged 36 (12, 117) months. The pathogen positive rate was 64.6% (166/257) and the in-hospital mortality was 27.6% (71/257). In the 208 pathogen-positive samples, bacteria was the most common (57.7%, 120/208) with G-negative bacteria predominating (55.8%, 67/120), followed by viruses (26.0%, 54/208). Nearly 99.2% (255/257) of the children were treated with antibacterial at the beginning, of whom 47.1% (121/257) were treated with carbapenems combined with vancomycin or linezolid. The proportion of 3 or more antibacterial combinations was higher in children with underlying diseases and hospital-acquired septic shock than in those without underlying disease or community-acquired septic shock (27.4% (49/179) vs. 14.1% (11/78), 29.4% (52/177) vs. 10.0% (8/80), χ2=5.35,11.56,all P<0.05). The proportion of initial combination of carbapenem and vancomycin or linezolid reduced from 52.5% (21/40) to 41.3% (19/46), and of adequate pathogen coverage increased from 40.0% (16/40) to 58.7% (27/46) in the last five years. Conclusions: The initial use of antibacterial drugs is common in children with septic shock in PICU, especially in those with hospital-acquired septic shock and underlying diseases. In recent years, antimicrobial combinations have decreased, but the pathogen coverage has improved, indicating that drug selection is more reasonable and accurate.

Cell membrane-coated human hair nanoparticles for precise disease therapies

Precision medicine is the ultimate goal for current disease therapies, including tumor and infection. The lack of specific targeted drugs for liver cancer and the lack of specific anti-infective drugs in the treatment of diabetic foot ulcer with infection (DFI) are the representative obstacles in those 2 major diseases currently plaguing human beings. Inventing natural biocompatible polymers derived from natural materials is one of the main development directions of current bio-medical materials. Though previous studies have demonstrated the potential application values of human black hair-derived nanoparticles (HNP) in cancer, methicillin-resistant Staphylococcus aureus (MRSA) infection, and thrombosis scenarios treatments, it still has not solved the problem of low local therapeutic concentration and general targeting ability. Here, we firstly modified the HNP with membrane encapsulations, which endowed these dual-pure natural bio-fabricated materials with better targeting ability at the disease sites with no reduction in photothermal therapy (PTT) effect. HNP coated by red blood cell membrane loaded with DSPE-PEG-cRGD peptide for the therapeutic application of liver cancer greatly prolonged in vivo circulation time and enhanced local targeting efficacy as well as low toxicity; HNP coated by the murine macrophage cell membrane (RAWM) for the DFIs treatment greatly promoted the adhesive ability of HNP on the bacteria and thereby improved the killing effect. Briefly, the appropriate cell membranes camouflaged HNP nanomedicine has the characteristics of excellent photothermal effect, an all-natural source with excellent biocompatibility and easy access, which is expected to have huge potential in both benign and malignant diseases.

Artificial intelligence for antiviral drug discovery in low resourced settings: A perspective

Current antiviral drug discovery efforts face many challenges, including development of new drugs during an outbreak and coping with drug resistance due to rapidly accumulating viral mutations. Emerging artificial intelligence and machine learning (AI/ML) methods can accelerate anti-infective drug discovery and have the potential to reduce overall development costs in Low and Middle-Income Countries (LMIC), which in turn may help to develop new and/or accessible therapies against communicable diseases within these countries. While the marketplace currently offers a plethora of data-driven AI/ML tools, most to date have been developed within the context of non-communicable diseases like cancer, and several barriers have limited the translation of existing tools to the discovery of drugs against infectious diseases. Here, we provide a perspective on the benefits, limitations, and pitfalls of AI/ML tools in the discovery of novel therapeutics with a focus on antivirals. We also discuss available and emerging data sharing models including intellectual property-preserving AI/ML. In addition, we review available data sources and platforms and provide examples for low-cost and accessible screening methods and other virus-based bioassays suitable for implementation of AI/ML-based programs in LMICs. Finally, we introduce an emerging AI/ML-based Center in Cameroon (Central Africa) which is currently developing methods and tools to promote local, independent drug discovery and represents a model that could be replicated among LMIC globally.

Advances of Cobalt Nanomaterials as Anti-Infection Agents, Drug Carriers, and Immunomodulators for Potential Infectious Disease Treatment.

Infectious diseases remain the most serious public health issue, which requires the development of more effective strategies for infectious control. As a kind of ultra-trace element, cobalt is essential to the metabolism of different organisms. In recent decades, nanotechnology has attracted increasing attention worldwide due to its wide application in different areas, including medicine. Based on the important biological roles of cobalt, cobalt nanomaterials have recently been widely developed for their attractive biomedical applications. With advantages such as low costs in preparation, hypotoxicity, photothermal conversion abilities, and high drug loading ability, cobalt nanomaterials have been proven to show promising potential in anticancer and anti-infection treatment. In this review, we summarize the characters of cobalt nanomaterials, followed by the advances in their biological functions and mechanisms. More importantly, we emphatically discuss the potential of cobalt nanomaterials as anti-infectious agents, drug carriers, and immunomodulators for anti-infection treatments, which might be helpful to facilitate progress in future research of anti-infection therapy.

An ultra sensitive and rapid SERS detection method based on vortex aggregation enhancement effect for anti-infective drug residues detection in water

The environmental pollution and human health risks caused by anti-infective residual drugs in the environment have attracted much attention. More convenient and effective detection methods to achieve the rapid and high sensitivity detection for such pollutants are required. In this work, a novel surface-enhanced Raman scattering (SERS) strategy based on vortex aggregation of AgNPs was proposed for the detection of anti-infective drugs in environmental water. The method enhanced the Raman signal of the targets by 2–7.4 times. The mechanism of aggregation enhancement effect under the low-frequency oscillation procedure which significantly enhanced the SERS signal of targets molecular on the aggregated AgNPs was revealed by UV–vis and ICP-MS methods. Three drugs of cefazolin sodium, pefloxacin, and chloroquine phosphate were determined. The detect limits were 3.97 × 10−9 mol/L, 2.42 × 10−10 mol/L, and 7.34 × 10−9 mol/L for cefazolin sodium, pefloxacin, and chloroquine phosphate, respectively. The quantitative relationships were obtained in a wide linear range of 4–5 orders as well as good accuracy and stability with the recoveries of 84.0%–97.1% and the relative standard deviations (RSDs) less than 4.6% for spiked in actual water samples. This method also had excellent repeatability and stability, which have potential application for rapid detection of trace pollutants in water environment.

Indolin-2-one Nitroimidazole Antibiotics Exhibit an Unexpected Dual Mode of Action.

Nitroimidazoles such as metronidazole are used as anti-infective drugs against anaerobic bacteria. Upon in vivo reduction of the nitro group, reactive radicals damage DNA and proteins in the absence of oxygen. Unexpectedly, a recent study of nitroimidazoles linked to an indolin-2-one substituent revealed potent activities against aerobic bacteria. This suggests a different, yet undiscovered mode of action (MoA). To decipher this MoA, we first performed whole proteome analysis of compound-treated cells, revealing an upregulation of bacteriophage-associated proteins, indicative of DNA damage. Since DNA binding of the compound was not observed, we applied activity-based protein profiling (ABPP) for direct target discovery. Labeling studies revealed topoisomerase IV, an essential enzyme for DNA replication, as the most enriched hit in pathogenic Staphylococcus aureus cells. Subsequent topoisomerase assays confirmed the inhibition of DNA decatenation in the presence of indolin-2-one nitroimidazole with an activity comparable to ciprofloxacin, a known inhibitor of this enzyme. Furthermore, we determined significantly increased redox potentials of indolin-2-one nitroimidazoles compared to classic 5-nitroimidazoles such as metronidazole, which facilitates in vivo reduction. Overall, this study unravels that indolin-2-one-functionalized nitroimidazoles feature an unexpected dual MoA: first, the direct inhibition of the topoisomerase IV and second the classic nitroimidazole MoA of reductive bioactivation leading to damaging reactive species. Importantly, this dual MoA impairs resistance development. Given the clinical application of this compound class, the new mechanism could be a starting point to mitigate resistance.

Simultaneous Determination of 37 Anti-infective Drugs Potentially Illegally Added to Cosmetics that Claimed to have Anti-acne Effects by UHPLC-Q-TOF HRMS

Simultaneous Determination of 37 Anti-infective Drugs Potentially Illegally Added to Cosmetics that Claimed to have Anti-acne Effects by UHPLC-Q-TOF HRMS

Titanium surface polyethylene glycol hydrogel and gentamicin-loaded cross-linked starch microspheres release system for anti-infective drugs

Objective To design and construct a hydrogel drug-controlled release system loaded with gentamicin on a titanium surface, and to evaluate the in vitro drug release behaviour and antibacterial properties and biocompatibility of the controlled release system. Methods Titanium (Ti) surface was coated with poly dopamine (PDA) substrate, and then polyethylene glycol (PEG) was attached to PDA. The composite drug microsphere controlled release layer formed by gentamicin (GEN) and cross-linked starch (CSt) were subsequently covered with poly lactic⁃co⁃glycolic acid (PLGA) as a barrier to construct a Ti-GEN-Cst-PLGA anti-infective drug controlled release system. Results The hydrogel drug release system was successfully constructed. The results of in vitro anti-staphylococcus aureus (SAU) assay, anti-staphylococcus epidermidis (SEP) assay and anti-Escherichia coli (ECO) assay showed that Ti-GEN-Cst-PLGA could effectively inhibit the growth of three bacteria. Assay in the New Zealand rabbit found that Ti-GEN-Cst-PLGA could promote wound healing at the 3rd week after implantation, and the pathology assay found that the Ti-GEN-Cst-PLGA group had less inflammatory reactions and significant tissue proliferation at the endophyte contact surface. Conclusion Ti-GEN-Cst-PLGA can effectively inhibit the inflammatory response and promote wound healing, or may be a potential treatment for orthopaedic endophytes.

Antibacterial and Phagocytic Properties of a Novel Silver-Interferon Complex

Background: Developing new antibacterial and antiviral drugs are considered a significant issue due to the emergence and spread of resistant strains of microorganisms. The COVID-19 pandemic has dramatically increased the need for new broad-spectrum anti-infective agents. Objective: This experimental study aimed to investigate the antibacterial and phagocytic properties of silver-interferon preparation. The combination of properties of complex drugs makes them promising for treating drug-resistant infections and bacterial complications of viral diseases. Method: The antibacterial effect of the silver-interferon platform was investigated by agar diffusion and serial dilution methods. The drug's effect on the functional activity of phagocytes was studied on human neutrophils in a Staphylococcus aureus uptake test. Results: Investigations have shown that the silver-interferon complex possesses a bactericidal mechanism of action against tested bacterial strains, including Streptococcus pneumonia, Salmonella enteritidis, Staphylococcus aureus, Escherichia coli. Streptococcus pneumonia was the most susceptible bacterial target for the tested complex, with a growth inhibition zone of 12±0.6 mm and a minimal bactericidal concentration of 0.08 mg/ml. A slight stimulating action of the drug in relation to the activity of phagocytes was revealed. Conclusion: Silver-interferon has proved as a prospective anti-infective drug with a wide range of activities.

Integrated Genotoxicity Testing of three anti-infective drugs using the TGx-DDI transcriptomic biomarker and high-throughput CometChip® assay in TK6 cells.

Genotoxicity testing relies on the detection of gene mutations and chromosome damage and has been used in the genetic safety assessment of drugs and chemicals for decades. However, the results of standard genotoxicity tests are often difficult to interpret due to lack of mode of action information. The TGx-DDI transcriptomic biomarker provides mechanistic information on the DNA damage-inducing (DDI) capability of chemicals to aid in the interpretation of positive in vitro genotoxicity data. The CometChip® assay was developed to assess DNA strand breaks in a higher-throughput format. We paired the TGx-DDI biomarker with the CometChip® assay in TK6 cells to evaluate three model agents: nitrofurantoin (NIT), metronidazole (MTZ), and novobiocin (NOV). TGx-DDI was analyzed by two independent labs and technologies (nCounter® and TempO-Seq®). Although these anti-infective drugs are, or have been, used in human and/or veterinary medicine, the standard genotoxicity testing battery showed significant genetic safety findings. Specifically, NIT is a mutagen and causes chromosome damage, and MTZ and NOV cause chromosome damage in conventional in vitro tests. Herein, the TGx-DDI biomarker classified NIT and MTZ as non-DDI at all concentrations tested, suggesting that NIT’s mutagenic activity is bacterial specific and that the observed chromosome damage by MTZ might be a consequence of in vitro test conditions. In contrast, NOV was classified as DDI at the second highest concentration tested, which is in line with the fact that NOV is a bacterial DNA-gyrase inhibitor that also affects topoisomerase II at high concentrations. The lack of DNA damage for NIT and MTZ was confirmed by the CometChip® results, which were negative for all three drugs except at overtly cytotoxic concentrations. This case study demonstrates the utility of combining the TGx-DDI biomarker and CometChip® to resolve conflicting genotoxicity data and provides further validation to support the reproducibility of the biomarker.

Harmine, an inhibitor of the type III secretion system of Salmonella enterica serovar Typhimurium.

New therapeutic strategies for clinical Salmonella enterica serovar Typhimurium (S. Typhimurium) infection are urgently needed due to the generation of antibiotic-resistant bacteria. Inhibition of bacterial virulence has been increasingly regarded as a potential and innovative strategy for the development of anti-infection drugs. Salmonella pathogenicity island (SPI)-encoded type III secretion system (T3SS) represents a key virulence factor in S. Typhimurium, and active invasion and replication in host cells is facilitated by the secretion of T3SS effector proteins. In this study, we found that harmine could inhibit T3SS secretion; thus, its potential anti-S. Typhimurium infection activity was elucidated. Harmine inhibits the secretion and expression of T3SS effector proteins and consequently attenuates the S. Typhimurium invasion function of HeLa cells. This inhibition may be implemented by reducing the transcription of pathogenesis-related SPI-1 transcriptional activator genes hilD, hilC, and rtsA. Harmine improves the survival rate and bacterial loads of mice infected with S. Typhimurium. In summary, harmine, an effective T3SS inhibitor, could be a leading compound for the development of treatments for Salmonella infection.

DEVELOPMENT AND VALIDATION OF STABILITY INDICATING HPTLC METHOD FOR THE SIMULTANEOUS ESTIMATION OF TINIDAZOLE AND FLUCONAZOLE AND ITS APPLICABILITY IN MARKETED DOSAGE FORM

Objective: The current method is focused on the development and validation of a simple, rapid, precise and robust stability indicating High Performance Thin Layer Chromatography for the simultaneous estimation of anti-infective drugs Tinidazole and Fluconazole in bulk and its pharmaceutical dosage form. The method was tailored to analyse the drugs in their commercial dosage form (tablets) with no interference from ingredients. Methods: Chromatographic separation was performed over precoated TLC plates (60 F254, 20 cm × 10 cm, 250 μm thickness, Merck) via a linear ascending technique using toluene: acetonitrile as the mobile phase in the ratio 6:4 v/v. Detection and quantification was achieved at the isobestic point of the two drugs, which was observed at 263 nm through Spectro-densitometric analysis. Analytical performance of the proposed HPTLC method was validated according to the ICH guidelines with respect to the linearity, accuracy, precision, detection and quantitation limits, robustness and specificity. Results: Tinidazole and Fluconazole were well separated and identified with an Rf value of about 0.46±0.03 and 0.75±0.05, respectively. The calibration curves were linear over a concentration range of 800-1200ng/spot for Tinidazole and 60-90ng/spot for Fluconazole with correlation coefficients (r2) more than 0.998. The above-developed method was validated as per ICH guidelines Q2(R1) and was found to be precise, sensitive, accurate and robust. Conclusion: The validated stability indicating HPTLC method was found to be simple, precise, accurate and sensitive for the concurrent quantification of Tinidazole and Fluconazole in pharmaceutical dosage form and can be released into quality control for regular analysis.

Drug-induced liver injury after anti tuberculosis drugs administration, how to diagnose? a case report

Drug-induced liver injury (DILI) is a liver injury caused by various drugs, herbs, or other xenobiotics, which causes abnormalities in liver tests or liver dysfunction in the absence of other causes of liver damage. The most common causative drugs are antituberculosis drugs (ATDs), anti-infective drugs, and natural herbal medicines. The diagnosis of DILI can be difficult due to the lack of specific signs, symptoms and tests and is partly a diagnosis based on exclusion. In this case report, we will discuss how to diagnosis DILI TB and causative assessment using RUCAM score. A male, 64 years old, has complained of weakness since 1 week ago and worsened since 1 day ago. The patient also felt persistent nausea for 1 week, so his eating and drinking decreased. Besides, he complained getting abdominal pain, especially in the upper right region and heartburn. The patient has been on first category of TB treatment since 20 days ago. Chest X-ray showed Lung TB with infiltrate in multiple cavities. Abdominal ultrasound showed no abnormality. The patient was discharged from our hospital after 6 days of hospitalization. DILI remains a diagnosis of exclusion based primarily on a detailed history and judicious use of blood tests, hepatobiliary imaging, and liver biopsy. The Roussel Uclaf causality assessment method (RUCAM) system is an assigning point for clinical, biochemical, serologic and radiologic features of liver injury. We use RUCAM score to make an assessment that show the likelihood of the hepatic injury due to a specific medication.

Changes in Serum CRP and PCT Levels in Patients with Acute Simple Lower Urinary Tract Infection and Evaluation of the Efficacy of Treatment with Shuangdong Capsules.

Objective The aim of this study is to investigate the changes and significance of serum C-reactive protein (CRP) and procalcitonin (PCT) levels in patients with acute simple lower urinary tract infection (ALUTI) and to analyze the efficacy of treatment with Shuangdong capsules, so as to provide a basis for the rational clinical application of drugs. Methods 92 patients with ALUTI (observation group) were randomly divided into 46 cases each in group A and group B. Group A was treated with basic anti-infective drugs, while group B was treated with Shuangdong capsules, and the duration of treatment in both groups was 14 days. The curative effect of the two groups was analyzed, and the changes in serum PCT and CRP levels were compared before and after treatment and compared with 40 healthy people in the control group. Results The serum PCT and CRP levels, the number of urinary leukocyte count (LEU), and the number of urinary bacterial count (BACT) were significantly higher in Group A and Group B than in the healthy control group before treatment (P < 0.05). After treatment, the total clinical efficiency of patients in group B (97.83%) was significantly higher than that in group A (78.26%) (P < 0.05). The serum PCT and CRP levels, the number of urinary LEU, and the number of urinary BACT decreased in both groups after treatment compared with those before treatment, and all of them were significantly lower in group B than in group A (P < 0.05). The area under the curve (AUC) values of serum PCT and CRP levels for the diagnosis of acute simple lower urinary tract infection were 0.747 (95% CI 0.633–0.860) and 0.926 (95% CI 0.870∼0.982), both with high sensitivity and specificity. Conclusion The Shuangdong capsule combined with conventional antibacterial drugs has better clinical efficacy in the treatment of acute simple lower urinary tract infection; serum CRP and PCT levels in patients with acute simple lower urinary tract infection can be used as indicators for diagnosis and efficacy determination of the infection.