Pharmacy students’ knowledge and attitudes about antibiotics in Kosovo

Objective: The main objective of this study was to assess the knowledge and attitude among Pharmacy students of the University of Prishtina in regards to the antibiotics. Methods: 144 pharmacy students at the University of Prishtina were recruited in this study to complete a self-administered questionnaire. The total number of questions in this questionnaire was eight (8), covering two (2) major themes: self-report of the current and past antibiotic use and behavior; and anticipated prescription behavior of antibiotics upon graduation. The data was statistically analyzed through using SPSS for Windows. Descriptive analysis was employed, and the results were expressed in frequency and percentages. Results: The results showcased a good knowledge of antibiotic among students. The most common answer of students' knowledge about antibiotics was good or moderate (82 %), while 63.2% of the subjects used antibiotics by self-decision, most of them (45 %) for sore throat. Upon graduation, 56.9 % of the students will not sell antibiotics without prescription and 85.4% think that module for rational use of antibiotics is very necessary to be inside the pharmacy syllabus. Conclusion: The study showed good and moderate knowledge of pharmacy students regarding the antibiotics. Half of them use antibiotics by self-decision but the majority of them stated that they will not serve the antibiotics without medical prescription. Specific modules and training for proper antibiotic use should be implemented within the Pharmacy program in The Faculty of Medicine.

Glycosylation and Glycodiversification in Polyketide Antibiotics: Unraveling Biosynthetic Steps in Nogalamycin Formation

Soil-dwelling Streptomyces bacteria are known for their ability to produce biologically active compounds such as antimicrobial, immunosuppressant, antifungal and anticancer drugs. S. nogalater is the producer of nogalamycin, a potential anticancer drug exhibiting high cytotoxicity and activity against human topoisomerases I and II. Nogalamycin is an anthracycline polyketide comprising a four-ring aromatic backbone,a neutral deoxy sugar at C7, and an amino sugar attached via an O–C bond at C1 and a C–C bond between C2 and C5´´. This kind of attachment of the amino sugar is unusual thus making the structure of the compound highly interesting. The sugar is also associated with the biological activity of nogalamycin, as it facilitates binding to DNA. Furthermore, the sugar moieties of anthracyclines are often crucial for their biological activity. Together the interesting attachment of the amino sugar and the general reliance of polyketides on the sugar moieties for bioactivity have made the study of the biosynthesis of nogalamycin attractive. The sugar moieties are typically attached by glycosyltransferases, which use two substrates: the donor and the acceptor. The literature review of the thesis is focused on the glycosylation of polyketides and the possibilities to alter their glycosylation patterns. My own thesis work revolves around the biosynthesis of nogalamycin. We have elucidated the individual steps that lead to its rather unique structure. We reconstructed the whole biosynthetic pathway in the heterologous host S. albus using a cosmid and a plasmid. In the process, we were able to isolate new compounds when the cosmid, which contains the majority of the nogalamycin gene cluster, was expressed alone in the heterologous host. The new compounds included true intermediates of the pathway as well as metabolites, which were most likely altered by the endogenous enzymes of the host. The biological activity of the most interesting new products was tested against human topoisomerases I and II, and they were found to exhibit such activities. The heterologous expression system facilitated the generation of mutants with inactivated biosynthetic genes. In that process, we were able to identify the functions of the glycosyltransferases SnogE and SnogD, solve the structure of SnogD, discover a novel C1-hydroxylase system comprising SnoaW and SnoaL2, and establish that the two homologous non-heme α-ketoglutarate and Fe2+ dependent enzymes SnoK and SnoN catalyze atypical reactions on the pathway. We demonstrated that SnoK was responsible for the formation of the additional C–C bond, whereas SnoN is an epimerase. A combination of in vivo and in vitro techniques was utilized to unravel the details of these enzymes. Protein crystallography gave us an important means to understand the mechanisms. Furthermore, the solved structures serve as platforms for future rational design of the enzymes.

Prophylactic antibiotics for the prevention of cellulitis

Background: Cellulitis of the lower leg accounts for 2-3% of hospital admissions (Cox et al, 1998), with an average length of in-patient stay of nine days. Studies have reported that up to half of these patients suffer further episodes (Cox et al, 1998; Dupuy et al, 1999). Reducing the recurrence of cellulitis could therefore have a significant impact on both patient morbidity and NHS costs. Aims: To assess whether prophylactic antibiotics prescribed after an episode of cellulitis of the leg results in fewer subsequent attacks and reduced health service costs (PATCH prophylactic antibiotics for the treatment of cellulitis at home). Methods: This article describes two related studies in which participants are randomised to receive either 12 months of prophylaxis (PATCH I) or six months of prophylaxis (PATCH II). The PATCH I study recruits only patients with recurrent disease defined as two or more episodes of cellulitis in the last three years. PATCH II has more open criteria and includes patients with a first episode of cellulitis and also participants with recurrent disease. It is expected that 260 patients will be recruited into PATCH I and 400 patients into PATCH II.

An Amidinohydrolase Provides the Missing Link in the Biosynthesis of Amino Marginolactone Antibiotics

Desertomycin A is an aminopolyol polyketide containing a macrolactone ring. We have proposed that desertomycin A and similar compounds (marginolactones) are formed by polyketide synthases primed not with gamma-aminobutanoyl-CoA but with 4-guanidinylbutanoyl-CoA, to avoid facile cyclization of the starter unit. This hypothesis requires that there be a final-stage de-amidination of the corresponding guanidino-substituted natural product, but no enzyme for such a process has been described. We have now identified candidate amidinohydrolase genes within the desertomycin and primycin clusters. Deletion of the putative desertomycin amidinohydrolase gene dstH in Streptomyces macronensis led to the accumulation of desertomycin B, the guanidino form of the antibiotic. Also, purified DstH efficiently catalyzed the in vitro conversion of desertomycin B into the A form. Hence this amidinohydrolase furnishes the missing link in this proposed naturally evolved example of protective-group chemistry.

Antibiotics resistance of Stenotrophomonas maltophilia strains isolated from various clinical specimens.

Background: A limited number of antibiotics are recommended for the therapy of Stenotrophomonas maltophilia infections due to therapy difficulties caused by its numerous mechanisms of resistance. Objectives: In this study conducted over a period of approximately 5 years we aimed to determine resistance rates of S. maltphilia based on drug classification recommended by Clinical and Laboratory Standards Institute. Methods: A total of 118 S. maltphilia strains isolated from various clinical specimens between January 2006 and June 2012 were included in the study. BD Phoenixautomated microbiology system (Becton Dickinson, USA) was utilized for species level identification and antibiotic susceptibility testing. Results: Sixty seven of S. maltphilia strains were isolated from tracheal aspirate isolates, 17 from blood, 10 from sputum, 10 from wound and 14 from other clinical specimens. Levofloxacin was found to be the most effective antibiotic against S. maltphilia strains with resistance rate of 7.6%. The resistance rates to other antibiotics were as follows: chloramphenicol 18.2%, trimethoprim-sulfamethoxazole 20.3% and ceftazidime 72%. Conclusion: The study revealed that S. maltphilia is resistant to many antibiotics. The treatment of infections caused by S. maltphilia should be preferred primarily as levofloxacin, chloramphenicol, and TMP-SXT, respectively.

Feasibility study to inform the design of a UK multi-centre randomised controlled trial of prophylactic antibiotics for the prevention of recurrent cellulitis of the leg

Background: This paper describes the results of a feasibility study for a randomised controlled trial (RCT). Methods: Twenty-nine members of the UK Dermatology Clinical Trials Network (UK DCTN) expressed an interest in recruiting for this study. Of these, 17 obtained full ethics and Research & Development (R&D) approval, and 15 successfully recruited patients into the study. A total of 70 participants with a diagnosis of cellulitis of the leg were enrolled over a 5-month period. These participants were largely recruited from medical admissions wards, although some were identified from dermatology, orthopaedic, geriatric and general surgery wards. Data were collected on patient demographics, clinical features and willingness to take part in a future RCT. Results: Despite being a relatively common condition, cellulitis patients were difficult to locate through our network of UK DCTN clinicians. This was largely because patients were rarely seen by dermatologists, and admissions were not co-ordinated centrally. In addition, the impact of the proposed exclusion criteria was high; only 26 (37%) of those enrolled in the study fulfilled all of the inclusion criteria for the subsequent RCT, and were willing to be randomised to treatment. Of the 70 participants identified during the study as having cellulitis of the leg (as confirmed by a dermatologist), only 59 (84%) had all 3 of the defining features of: i) erythema, ii) oedema, and iii) warmth with acute pain/tenderness upon examination. Twenty-two (32%) patients experienced a previous episode of cellulitis within the last 3 years. The median time to recurrence (estimated as the time since the most recent previous attack) was 205 days (95% CI 102 to 308). Service users were generally supportive of the trial, although several expressed concerns about taking antibiotics for lengthy periods, and felt that multiple morbidity/old age would limit entry into a 3-year study. Conclusion: This pilot study has been crucial in highlighting some key issues for the conduct of a future RCT. As a result of these findings, changes have been made to i) the planned recruitment strategy, ii) the proposed inclusion criteria and ii) the definition of cellulitis for use in the future trial.

How much do patients in Blantyre, Malawi know about antibiotics and other prescription only medicines?

Introduction This qualitative and quantitative exploratory study was conducted to assess patients’/customers’ knowledge, beliefs and practices about antibiotics and other prescription only medication (POM) in 10 community pharmacies in Blantyre, Malawi. Method 5 out of 10 pharmacies were selected by simple random sampling and 54 participants attending the selected pharmacies were interviewed. Results The major antibiotic drugs frequently requested without a doctor’s prescription were Bactrim (Cotrimoxazole) and amoxicillin. Knowledge of these medicines was attributed to the patients’ previous medical conditions and doctor’s prescription. However this knowledge does not extend to understanding of proper therapeutic indications and dosage regimens for antibiotic therapy. The majority (30 out of 54 participants) did not know about the importance of completing a prescribed course of antibiotics. All 54 participants believed in the efficiancy of antibiotics. Conclusion Study participants had wide range of knowledge about antibiotics and other POM from hospital, but the majority had limited knowledge regarding clinical indications and correct dosage schedules.

False identification of other microorganisms as Staphylococcus aureus in Southern Nigeria

Purpose: Staphylococcus aureus is the causative agent of many infections and the advent MRSA has drawn much attention to it. However, some organisms have been noted to be wrongly identified as S. aureus through phenotypic identifications leading to wrong treatment of infections. This study is therefore undertaken to evaluate the rate of false identification of other organisms as S. aureus in Southern Nigeria. Methods: 507 microorganisms which have been previously identified as S. aureus in 8 States in Southern Nigeria through characteristic morphology on blood agar, Gram staining, growth and fermentation on Mannitol Salt Agar and coagulase formation were collected. All the isolates were identified in this study through sequencing of 16S rRNA and detection of spa gene. The percentages of true and false identities were determined. Results: Of the 507 isolates previously identified as S. aureus, only 54 (11 %) were confirmed as S. aureus while the rest were coagulase negative Staphylococci (85 % misidentification rate), Bacillus sp. (12 % misidentification rate), and Brevibacterium sp. (3 % misidentification rate). Conclusion: A high rate of false positive identification of S. aureus which could lead to the misuse of antibiotics in emergency situation has been identified in this study. The use of standard methods for the identification of S. aureus at all times is highly recommended.

Parental views of antibiotic use in children with upper respiratory tract infections in Jordan

Purpose: To assess the knowledge, attitudes and practices of parents towards antibiotics use for upper respiratory tract infections (URTIs) in Jordan. Methods: A cross-sectional study was carried out at 10 private outpatients’ pediatric clinics across Amman-Jordan from September to December 2013. During the study period, 1329 parents of young children who fulfilled the inclusion criteria and agreed to participate were interviewed, and completed a validated structured questionnaire. Results: A large proportion of parents (903, 68 %) believed that weather change was the main cause of acute URTIs in their children. Although 1098 (82.8 %) of parents were aware that the recurrent use of antibiotics leads to a decrease in effectiveness due to bacterial resistance, 859 (64.6 %) of the respondents reported that they would give antibiotics without prescription. Fathers (135, 40.2 %), were significantly more aware that URTIs follow its natural course without antibiotic administration compared to mothers (N = 327, 32.9 %), respectively (p = 0.005). Conclusion: There is a lack of adequate parental knowledge concerning the use and misuse of antibiotics in children in Jordan. National publicity campaign should be mounted to improve awareness. Furthermore, existing laws should be enforced to prevent parents from purchasing antibiotics over-thecounter (OTC).

Assessment of the Occurrence and Potential Risks of Antibiotics and their Metabolites in South Florida Waters Using Liquid Chromatography Tandem Mass Spectrometry

An automated on-line SPE-LC-MS/MS method was developed for the quantitation of multiple classes of antibiotics in environmental waters. High sensitivity in the low ng/L range was accomplished by using large volume injections with 10-mL of sample. Positive confirmation of analytes was achieved using two selected reaction monitoring (SRM) transitions per antibiotic and quantitation was performed using an internal standard approach. Samples were extracted using online solid phase extraction, then using column switching technique; extracted samples were immediately passed through liquid chromatography and analyzed by tandem mass spectrometry. The total run time per each sample was 20 min. The statistically calculated method detection limits for various environmental samples were between 1.2 and 63 ng/L. Furthermore, the method was validated in terms of precision, accuracy and linearity. The developed analytical methodology was used to measure the occurrence of antibiotics in reclaimed waters (n=56), surface waters (n=53), ground waters (n=8) and drinking waters (n=54) collected from different parts of South Florida. In reclaimed waters, the most frequently detected antibiotics were nalidixic acid, erythromycin, clarithromycin, azithromycin trimethoprim, sulfamethoxazole and ofloxacin (19.3-604.9 ng/L). Detection of antibiotics in reclaimed waters indicates that they can’t be completely removed by conventional wastewater treatment process. Furthermore, the average mass loads of antibiotics released into the local environment through reclaimed water were estimated as 0.248 Kg/day. Among the surface waters samples, Miami River (reaching up to 580 ng/L) and Black Creek canal (up to 124 ng/L) showed highest concentrations of antibiotics. No traces of antibiotics were found in ground waters. On the other hand, erythromycin (monitored as anhydro erythromycin) was detected in 82% of the drinking water samples (n.d-66 ng/L). The developed approach is suitable for both research and monitoring applications. Major metabolites of antibiotics in reclaimed wates were identified and quantified using high resolution benchtop Q-Exactive orbitrap mass spectrometer. A phase I metabolite of erythromycin was tentatively identified in full scan based on accurate mass measurement. Using extracted ion chromatogram (XIC), high resolution data-dependent MS/MS spectra and metabolic profiling software the metabolite was identified as desmethyl anhydro erythromycin with molecular formula C36H63NO12 and m/z 702.4423. The molar concentration of the metabolite to erythromycin was in the order of 13 %. To my knowledge, this is the first known report on this metabolite in reclaimed water. Another compound acetyl-sulfamethoxazole, a phase II metabolite of sulfamethoxazole was also identified in reclaimed water and mole fraction of the metabolite represent 36 %, of the cumulative sulfamethoxazole concentration. The results were illustrating the importance to include metabolites also in the routine analysis to obtain a mass balance for better understanding of the occurrence, fate and distribution of antibiotics in the environment. Finally, all the antibiotics detected in reclaimed and surface waters were investigated to assess the potential risk to the aquatic organisms. The surface water antibiotic concentrations that represented the real time exposure conditions revealed that the macrolide antibiotics, erythromycin, clarithromycin and tylosin along with quinolone antibiotic, ciprofloxacin were suspected to induce high toxicity to aquatic biota. Preliminary results showing that, among the antibiotic groups tested, macrolides posed the highest ecological threat, and therefore, they may need to be further evaluated with, long-term exposure studies considering bioaccumulation factors and more number of species selected. Overall, the occurrence of antibiotics in aquatic environment is posing an ecological health concern.

Revisión sistemática de la literatura de las intervenciones diseñadas para el comportamiento de automedicación no responsable

La automedicación no responsable se ha convertido en un problema de salud pública global en las últimas décadas, por sus consecuencias individuales (por ejemplo, la intoxicación) y colectivas (por ejemplo, la resistencia microbiana a los antibióticos). Las intervenciones orientadas a este comportamiento han sido aisladas y muy diferentes. Aunque se tiene evidencia de que su aplicación puede traer beneficios en diferentes poblaciones, no se halló en la literatura una compilación sistemática de dichas intervenciones. El objetivo de la presente revisión es sistematizar la literatura científica sobre las diferentes alternativas de intervención del comportamiento individual de automedicación no responsable. En cuanto al método, la revisión de literatura involucró la búsqueda sistemática de “automedicación” e “intervención” en las bases de datos académicas internacionales con contenidos de psicología, suscritas por la Biblioteca de la Universidad del Rosario. Como resultado se encontró que las intervenciones orientadas al comportamiento de automedicación no responsable se pueden clasificar en dos grandes grupos: (a) intervenciones regulatorias, con dirección “arriba hacia abajo”, que suponen una acción de los Estados nacionales por medio de sus legislaciones o de entidades internacionales (por ejemplo, Organización Mundial de la Salud); y (b) intervenciones educativas, con dirección “abajo hacia arriba”, que suponen acciones con individuos y comunidades con el fin de enseñar acerca del uso adecuado de los medicamentos. Se concluye acerca de la necesidad de complementar ambos tipos de intervención, los cuales, si bien demuestran resultados positivos, aisladamente son insuficientes para contrarrestar integralmente este fenómeno creciente y complejo.

Biochemical characterization of systemic bacteria in bananas, sensitivity to antibiotics and plant phytotoxicity during shoot proliferation.

2016

Role of organic acids and phytonutrients as natural alternatives to antibiotics in supporting intestinal functionality

The gastrointestinal tract (GIT) represents the major portion of the body that interfaces with the external environment, with the double function of food processing and line of defense of the body. Numerous components support and regulate the barrier function of the GIT, such as tight junctions (TJs), cytokines, commensal and pathogenic microorganisms, and other systems of the organism, as the endocannabinoid system (ECS). The ECS can control several gastrointestinal functions, as well as the regulation of intestinal inflammation. Failure of the intestinal barrier function triggers an increase of the concentration of pro-inflammatory cytokines and leads to a reduction in intestinal functionality. This thesis aimed to explore the potential of natural compounds as a new alternative approach to antibiotics not only as antimicrobial, but also supporting intestinal maturation and integrity, and as immune-boosting agents. Different experiments were performed to evaluate the potential of nature-identical compounds (NICs), organic acids (OAs), and essential oils (EOs) to support and fight various stressful stimuli. In vitro, a well characterized blend of NICs and OAs were able to improve TJs and transepithelial electrical resistance (TEER) in an intestinal cell line, exerting an anti-inflammatory potential. EOs enhanced TEER and TJs mRNA levels, with a reduction of paracellular permeability, showing antioxidant and antimicrobial properties. In vivo, thymol modulates the gene expression of ECS and gut chemosensing in the GIT of piglets, where the precise localization of the cannabinoid receptors was immunohistochemically confirmed, suggesting an anti-inflammatory potential. In conclusion, natural alternative molecules represent an effective alternative to support or replace the classical pharmacological prophylaxis. These alternative molecules act not only as antimicrobial agents, but also exerted a crucial role in supporting the intestinal barrier function, preventing oxidative stress, and reducing inflammation. Moreover, thymol seems able to modulate the ECS, representing a novel frontier to support animal health and productivity.

Extensive Frontoparietal Abscess: Complication Of Frontal Sinusitis (pott's Puffy Tumor).

First described in 1768, the Pott's puffy tumor is a subperiosteal abscess associated with frontal bone osteomyelitis, resulting from trauma or frontal sinusitis. The classic clinical presentation consists of purulent rhinorrhea, fever, headache, and frontal swelling. The diagnosis is confirmed by CT scan and treatment requires intravenous antibiotics, analgesia, and surgical intervention. Early diagnosis and aggressive medical and surgical approach are essential for a good outcome. It rare and the early diagnosis is important; we describe the case of a 14-year-old adolescent with Pott's puffy tumor who was initially treated inadequately, evolving with extensive frontoparietal abscess. The patient underwent surgical treatment with endoscopic endonasal and external approaches combined. Intravenous antibiotics were prescribed for a prolonged time, with good outcome and remission of the complaints.