Antibacterial Profile in vitro and in vivo of New 1,4-Naphthoquinones Tethered to 1,2,3-1H-Triazoles against the Planktonic Growth of Streptococcus mutans

The cariogenic processes are mainly caused by the bacterium Streptococcus mutans (S. mutans) and consist of the demineralization of the tooth that occurs when the acid production overcomes the natural repair or if a problem occurs in the last one. In this work, we performed the synthesis of twenty-one 1,4-naphthoquinones tethered to 1,2,3-1H-triazoles (8a-8k and 9a-9j), antibacterial evaluation against the S. mutans in vitro and the acute toxicity of the better ones in vivo. We observed strong inhibition results in the disc diffusion test ranging, the halos of inhibitions, from 18.66 (± 0.57) to 29 (± 2.64) mm, and good values in the minimum inhibitory concentration (5 to 50 μg), for the compounds 9e, 9h, 9i and 9j. Furthermore, they do not have a cytotoxic effect at the concentrations tested. Besides that, in the in vivo test, they show some slight alteration in the histopathological analyses and the biochemistry. Thus, we found four potential candidates to become instruments for the treatment of cavities.

Characterisation of AmpC / ESBL genes in some pathogen gram-negatives isolated from clinical cases of livestock and companion animals

This study was aimed to search and characterize the AmpC and/or ESBL genes of Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa isolated from clinical cases of local livestock and companion animals between 2017 and 2019. A total of eight ceftiofur-resistant E. coli (n= 7) and ceftiofur-resistant K. pneumoniae (n= 1) and seven P. aeruginosa were isolated from different cases in local animals. By combination disc method, six E. coli isolates and one K. pneumoniae isolate were found to be ESBL producers. By combination of the disc method and double disc synergy test, no P. aeruginosa isolates were found as ESBL producers. In the agar disc diffusion test (ADDT) performed with cefoxitin and cefoxitin-boronic, only one E. coli was determined as AmpC producer. In ESBL-producing isolates, only the CTX-M class gene was detected by polymerase chain reaction (PCR) and subsequent sequence analysis revealed CTX-M-3 and CTX-M-15 variants. An AmpC positive E. coli isolate was found to carry plasmidic ampC gene in cmy-2 variant from CIT family. It was observed that P. aeruginosa isolates did not carry the plasmidic ampC gene. After the chromosomal ampC gene of one P. aeruginosa was amplified by PCR and sequenced, R79Q and T105A mutations in the chromosomal ampC gene was revealed. This showed that overproduction of the ampC enzyme is involved in the resistance to β-lactams in P. aeruginosa isolates in the study.

Assessment of the Effect of Structural Modification of Ibuprofen on the Penetration of Ibuprofen from Pentravan® (Semisolid) Formulation Using Human Skin and a Transdermal Diffusion Test Model

The effect of transdermal vehicle (Pentravan®) on skin permeability was examined for unmodified ibuprofen (IBU) and ion pairs of ibuprofen with new L-valine alkyl esters [ValOR][IBU]. The percutaneous permeation across the human skin and transdermal diffusion test model (Strat-M® membranes) of ibuprofen and its structural modification were measured and compared using Franz diffusion cells. For comparison, the penetration of ibuprofen from a commercial product was also investigated. The cumulative amount of drug permeated through human skin at the end of the 24 h study was highest for ibuprofen derivatives containing propyl (C3), isopropyl (C3), ethyl (C2), and butyl (C4) esters. For Strat-M®, the best results were obtained with the alkyl chain length of the ester from C2 to C5. The permeation profiles and parameters were appointed, such as steady-state flux, lag time, and permeability coefficient. It has been shown that L-valine alkyl ester ibuprofenates, with the propyl, butyl, and amyl chain, exhibit a higher permeation rate than ibuprofen. The diffusion parameters of analyzed drugs through human skin and Strat-M® were similar and with good correlation. The resulting Pentravan-based creams with ibuprofen in the form of an ionic pair represent a potential alternative to other forms of the drug-containing analgesics administered transdermally. Furthermore, the Strat-M® membranes can be used to assess the permeation of transdermal preparations containing anti-inflammatory drugs.

Molecular characterization of enterohemorrhagic Escherichia coli isolated from diarrhea samples from human, livestock, and ground beef in North Jordan

Background and Aim: Enterohemorrhagic Escherichia coli (EHEC) is an important foodborne pathogen with worldwide distribution. Data regarding its presence, distribution, virulence, and antimicrobial susceptibility among various animal species and humans in Jordan are lacking. Therefore, the objectives of this study were to isolate and characterize EHEC from human and animal diarrhea fecal samples and ground beef samples. Materials and Methods: A total of 100 and 270 diarrhea fecal samples from humans and animals, respectively, were collected. In addition, 40 ground beef meat samples were collected from retail markets. EHEC was positively identified by detecting Shiga toxins (stx1 and stx2) genes using multiplex polymerase chain reaction (PCR). Antimicrobial susceptibility patterns were determined using the disk diffusion test. Beta-lactamase production was detected using the double disk diffusion test and the extended-spectrum beta-lactamases (ESBLs) were identified by detection of blaTEM, blaSHV, and OXA-1 genes using multiplex PCR. Pulsed-field gel electrophoresis (PFGE) was used to investigate the relatedness of EHEC isolates from different sources. Results: Out of 410 samples, 194 E. coli isolates were positively identified, of which 57 isolates (29%) were classified as EHEC. Thirty-five (61%) of EHEC isolates were serotyped as O157 (19: O157:H7 and 16: O157:NM). The stx1 gene was detected only among the sheep and goats isolates at a rate of 7.6% and 5.2%, respectively, while the stx2 gene was detected in only one ground beef meat sample. EHEC isolates showed high resistance patterns against amoxicillin, gentamycin, cephalexin, and doxycycline. Twenty-four out of 32 EHEC isolates were determined as ESBL producers, among which 14 isolates expressed the blaSHV gene and 19 isolates expressed the blaTEM while four expressed both genes. PFGE analysis revealed two clusters with high similarity (92%) originated from ground beef meat and cattle fecal samples. No similarities were found between human and animal E. coli isolates. Conclusion: Results of this study indicate widespread ESBL EHEC among humans, animals, and ground beef meat samples. These results represent an important alarm that requires the implementation of appropriate preventative measures by both human and animal health sectors to prevent the transmission of this important foodborne pathogen.

Evaluation of cefoxitin disc diffusion test as a rapid phenotypic method for detection of methicillin resistance

: Accuracy and promptness in the detection of methicillin resistance are of key importance in ensuring correct antibiotic treatment in infected patients and control of MR staphylococci in the hospital environment. The aim of this study was to detect MRSA phenotypically by oxacillin screen agar and Oxacillin MIC method and to evaluate cefoxitin disc diffusion test as a screening tool for MRSA detection. In the present study, a total of 50 isolates of from various clinical samples collected were used for the detection of Methicillin resistant (MRSA). Methicillin resistance was determined by oxacillin disc diffusion, cefoxitin disc diffusion the oxacillin screen agar test and MIC. Out of 50 isolates 21 (42%) isolates were detected as MRSA based on MIC method, which is considered as gold standard method for the detection of MRSA. All the isolates of MRSA were 100% susceptible to vancomycin and linezolid. rnIn the present study, cefoxitin diffusion method has given 100% sensitivity and specificity in concordance with MIC method. However, the oxacillin screen agar method showed 95.24% sensitivity and 96.55% specificity. As per our study and previous reports elsewhere on phenotypic detection of MRSA, cefoxitin is more potent inducer of the regulatory system and an accurate surrogate marker for the detection of MRSA in the routine susceptibility testing.

Respiratory muscle strength in patients after COVID-19

Respiratory muscles (RM) are a very important part of the respiratory system that enables pulmonary ventilation. This study aimed to assess the post-COVID-19 strength of RM by estimating maximum static inspiratory (MIP or PImax) and expiratory (MEP or PEmax) pressures and to identify the relationship between MIP and MEP and the parameters of lung function. We analyzed the data of 36 patients (72% male; median age 47 years) who underwent spirometry, and body plethysmography, diffusion test for carbon monoxide (DLCO) and measurement of MIP and MEF. The median time between the examinations and onset of COVID-19 was 142 days. The patients were divided into two subgroups. In subgroup 1, as registered with computed tomography, the median of the maximum lung tissue damage volume in the acute period was 27%, in subgroup 2 it reached 76%. The most common functional impairment was decreased DLCO, detected in 20 (55%) patients. Decreased MIP and MEP were observed in 5 and 11 patients, respectively. The subgroups did not differ significantly in MIP and MEP values, but decreased MIP was registered in the second subgroup more often (18%). There were identified no significant dependencies between MIP/MEP and the parameters of ventilation and pulmonary gas exchange. Thus, in patients after COVID-19, MIP and MEP were reduced in 14 and 31% of cases, respectively. It is reasonable to add RM tests to the COVID-19 patient examination plan in order to check them for dysfunction and carry out medical rehabilitation.

A Link Between Components of the Nasal Microbiome and an Individual’s Susceptibility to Influenza.

According to CDC influenza estimates, the flu infects ~40 million people and causes 24,000 to 60,000 deaths in the United States annually. Vaccination can be highly effective but is often a neglected tool for preventing infection. In this project, three methods were developed to compare an individual’s reported self-history of influenza infection to the types and amounts of nasal bacteria collected by nasal swab to assess if certain bacteria may correlate with less history of influenza infection. These three methods quantified species from four genera of bacteria - Staphylococcus, Micrococcus, Streptococcus and Haemophilus - and compared the amounts of each type of bacteria with participant survey answers regarding their history of influenza infection. In Method 1, a disk diffusion test with bacitracin distinguished isolates of Staphylococcus from Micrococcus. Higher ratios of Staphylococcus to Micrococcus were found in individuals less susceptible to influenza (p = 0.003). In Method 2, S. pyogenes and S. pneumoniae were distinguished based on their hemolytic patterns. A higher proportion S. pneumoniae significantly correlated with more history of influenza (p = 0.002). In Method 3, total numbers of Staphylococcus spp. and H. influenzae were compared. More frequent H. influenzae significantly correlated with higher influenza frequency (p = 0.006). While all three methods indicate correlations between specific nasal bacteria and influenza susceptibility, Method 2 was the simplest and least expensive to perform. Commercialization of one or more of these methods could result in a simple and inexpensive test to identify at-risk individuals for influenza.

ANTIMICROBIAL SUSCEPTIBILITY PATTERN OF STAPHYLOCOCCUS AUREUS STRAINS IN ISLAMABAD, PAKISTAN

Objective: To investigate the prevalence of S. aureus in hospitalized patients of Islamabad. Study Design: Cross-sectional study. Study Duration: Pakistan Institute of Medical Science, Applied Microbiology and Biotechnology Lab, COMSATS Institute of Information Technology, Islamabad, from Sep 2017 to Sep 2018. Methodology: A total of 500 samples were collected. The isolates were divided into four study groups according to their source of origin i.e. group 1 (dermal group), group 2 (nasal group), group 3 (blood group) and group 4 (urine group). Gram staining, catalase test and DNA se media analysis were done for validation of S. aureus. Disc diffusion test (for antibiotic susceptibility), Oxacillin disc test (to differentiate between methicillin-resistant Staphylococcus aureus and methicillin-susceptible staphylococcus aureus) and minimal inhibitory concentration (for susceptibility to vancomycin), were performed. Results: Degree of the prevalence of staphylococcus aureus was 21%, 17%, 9% and 8% in group 1, 2, 3 & 4 respectively. The overall prevalence of staphylococcus aureus was 19.5% in all isolates. The disc diffusion test showed the descending resistance pattern of isolates i.e. 100, 94, 94, 76, 58, 55, 47, 43, 40 and 37% for penicillin, ciprofloxacin, Kanamycin, erythromycin, tetracycline, oxazolidinone, sulfamethoxazole, doxycycline, clindamycin, and cipoxin respectively. Minimal inhibitory concentration found only one sample resistant at 2ug/l concentration of Vancomycin. Moreover, Oxacillin disc test showed 52% methicillin-susceptibleStaphylococcus aureus while 48.2% methicillin-resistant staphylococcus aureus among all isolates. Conclusion: There is an increase in the frequency of methicillin-resistant staphylococcus aureus. Single vancomycin resistant staphylococcus aureus strain was also isolated.