Rapid Detection of Carbapenemase-Producing Multidrug-Resistant (MDR) Pathogens by Modified Carba NP Test in Ventilator-Associated Pneumonia (VAP) in Elderly Patients.

Background Ventilator-associated pneumonia (VAP) is defined as pneumonia that develops 48 hours or more after endotracheal intubation or tracheostomy and is brought on by infectious organisms that are not present or incubating during mechanical ventilation. Multidrug-resistant organisms originate primarily from the hospital environment and significantly contribute to ventilator-associated pneumonia. These organisms pose a severe threat, leading to a higher mortality rate due to their resistance to more potent antibiotics. Methods The study aims to assess the efficacy of the modified Carba NP test in detecting carbapenemase-producing bacteria in geriatric VAP patients. Results Forty (38 gram-negative and 2 gram-positive) pathogens were isolated from VAP patients. The isolates were identified using standard laboratory protocol; Acinetobacter spp. (n=16; 40% ), followed by Klebsiella pneumoniae (n=13; 32.5%), is the most common organism isolated. Seventeen (44.73%) were multi-drug resistant gram-negative bacteria. The carbapenemase producers were detected by the Kirby-Bauer disc diffusion method and compared with the modified Carba NP test with a turnaround time of 12-18 hrs in comparison to the disk diffusion test which requires additional 12hrs. Carbapenemase production was seen in 12 (70.59%) MDR isolates (7-Acinetobacter spp, 3-Klebsiella pneumonia, 1-Escherichia coli, and 1-Pseudomonas aeruginosa).  Conclusion Modified Carba NP can be used as a rapid test to detect carbapenemase production, and it can replace the traditional disk diffusion method of detecting carbapenemase production. This test plays a crucial role in the management of critical patients by saving 12-18 hours to determine the most appropriate and effective antibiotic treatment. This timely decision is essential in preventing sepsis caused by localized infections.

The Effect of Speed and Rotation for Protaper File Systems on Postobturation Pain in a Single Visit and Multiple (Two) Visits in Root Canal Therapy: An In Vivo Study.

INTRODUCTION: The basic idea of a root canal treatment is to alleviate the pain and heal the infection within the infected tooth, which can be resolved in a single visit or multiple visits in root canal therapy. Therefore, in this article, an in vivo comparison of single visits and multiple visits using different rotation and speed for two Protaper universal design file systems is done on the basis of time taken and incidence of pain. MATERIALS AND METHODS: One hundred single-rooted premolars with irreversible symptomatic pulpitis were assigned to two groups of 50 patients each using the odd-even method, GA to be treated endodontically in a single visit and GB to be treated endodontically in multiple visits. Each group was further divided into two subgroups of 25 patients each on the basis of two different variations of speed and rotation for two Protaper file systems of the same design, GA1 (Hand Protapers) and GA2 (Rotary Protapers), GB1 (Hand Protapers) and GB2 (Rotary Protapers), respectively. After proper biomechanical shaping and cleaning, obturation was done with Gutta-percha cones and Ah plus sealer using Fast Pack obturation pen for warm vertical compaction. The pain was measured by a 100 mm modified visual analogue scale, and time was measured using a stopwatch. RESULTS: At 6-h intervals, post-obturation pain was more in single-visit root canal therapy than multiple-visit root canal therapy (P < 0.01). Single-visit rotary Protaper (GA2) had less incidence of post-obturation pain as compared with single-visit hand protapers (GA1) (P < 0.05). There was no significant difference in post-obturation pain in the multiple-visit hand protaper subgroup (GB1) and multiple-visit rotary protaper subgroup (GB2) (P > 0.05). Preoperative pain significantly influences the post-obturation pain. The statistical analysis was done using SPSS (Statistical Package for Social Sciences) version 17.0 statistical Analysis Software. CONCLUSIONS: The presence of preoperative pain can significantly influence the presence of postoperative pain. Most of the pain in both single-visit and multiple-visits root canal therapy occurred in the first 48 h after obturation, which decreases thereafter. Single-visit rotary protaper (GA2) had less incidence of post-obturation pain as compared with single-visit hand protapers (GA1). There was no significant difference in post-obturation pain in the multiple-visit hand protaper subgroup (GB1) and multiple-visit rotary protaper subgroup (GB2). Presence of sealer puff influences the duration of pain.

Healthcare Access, Pregnancy Intention, and Contraceptive Practices Among Reproductive-Aged Women Receiving Opioid Agonist Therapy in Northeast Tennessee.

OBJECTIVES: Women with substance use disorders often experience unique challenges to obtaining contraception and adhering to user-dependent methods. As a result, this at-risk population of women tends to have higher than average rates of unintended pregnancy. The objective of this study was to describe contraceptive use, pregnancy intentions, and adequacy of access to reproductive healthcare among women receiving opioid agonist therapy in northeast Tennessee. METHODS: A cross-sectional survey was piloted among female patients aged 18 to 55 years from two opioid agonist therapy clinics. Descriptive analyses were conducted using logistic regression to evaluate the statistical significance of bivariate associations. RESULTS: Of 91 participants, 84% reported having health insurance, with 70% perceiving having adequate access to health care. More than half had a history of unwanted pregnancy (53%), among whom few (23.1%) reported the consistent use of contraception at time of conception. Although most desired to avoid pregnancy (90%), only 59% of women reported the current use of regular contraception. Most of those not using regular contraception believed that they were not at risk for pregnancy (54.3%). CONCLUSIONS: Although most participants reported adequate access to health care and a desire to avoid pregnancy, few reported the consistent use of regular contraception. Furthermore, misperceptions regarding pregnancy risk were common among participants. Research is needed to identify barriers to contraceptive acceptance and causes of pregnancy risk misperceptions in this population of women at increased risk of unintended pregnancy.

Gene editing and genetic engineering approaches for advanced probiotics: A review.

The applications of probiotics are significant and thus resulted in need of genome analysis of probiotic strains. Various omics methods and systems biology approaches enables us to understand and optimize the metabolic processes. These techniques have increased the researcher's attention towards gut microbiome and provided a new source for the revelation of uncharacterized biosynthetic pathways which enables novel metabolic engineering approaches. In recent years, the broad and quantitative analysis of modified strains relies on systems biology tools such as in silico design which are commonly used methods for improving strain performance. The genetic manipulation of probiotic microorganisms is crucial for defining their role in intestinal microbiota and exploring their beneficial properties. This review describes an overview of gene editing and systems biology approaches, highlighting the advent of omics methods which allows the study of new routes for studying probiotic bacteria. We have also summarized gene editing tools like TALEN, ZFNs and CRISPR-Cas that edits or cleave the specific target DNA. Furthermore, in this review an overview of proposed design of advanced customized probiotic is also hypothesized to improvise the probiotics.

Metabolic Engineering for Probiotics and their Genome-Wide Expression Profiling.

Probiotic supplements in food industry have attracted a lot of attention and shown a remarkable growth in this field. Metabolic engineering (ME) approaches enable understanding their mechanism of action and increases possibility of designing probiotic strains with desired functions. Probiotic microorganisms generally referred as industrially important lactic acid bacteria (LAB) which are involved in fermenting dairy products, food, beverages and produces lactic acid as final product. A number of illustrations of metabolic engineering approaches in industrial probiotic bacteria have been described in this review including transcriptomic studies of Lactobacillus reuteri and improvement in exopolysaccharide (EPS) biosynthesis yield in Lactobacillus casei LC2W. This review summaries various metabolic engineering approaches for exploring metabolic pathways. These approaches enable evaluation of cellular metabolic state and effective editing of microbial genome or introduction of novel enzymes to redirect the carbon fluxes. In addition, various system biology tools such as in silico design commonly used for improving strain performance is also discussed. Finally, we discuss the integration of metabolic engineering and genome profiling which offers a new way to explore metabolic interactions, fluxomics and probiogenomics using probiotic bacteria like Bifidobacterium spp and Lactobacillus spp.

An overview of advanced technologies for selection of probiotics and their expediency: A review.

It has become easy to identify and select an appropriate microorganism with the advancement in various molecular biology and analytical techniques. The majority of the novel techniques are being implemented for the identification and characterization of microorganisms used for probiotic application. Standard microbial techniques such as biochemical testing and culture techniques routinely used for probiotic microbes screening, identification and selection. However, these standard techniques may not give complete information on the microbes that can be used for probiotic production. Furthermore, alternative molecular and analytical techniques such as 16S and 23S ribosomal DNA sequencing, RNA analysis by reverse transcriptase (RT-PCR), fluorescent in situ hybridization (FISH), quantitative analysis by real time PCR (RT-PCR or qPCR) and fluorescent activated cell sorting (FACS) are potentially used to confirm and select all types of lactic acid microorganism. All these approaches can be employed in the screening and selection of appropriate lactic acid bacteria which can be potentially used for the production of human use probiotics in large scale fermentation. This review mainly focuses on various tools and techniques used for effective screening and selection of a better candidate bacterium for probiotic applications.

Bioengineering for Microbial Inulinases: Trends and Applications.

Inulinase has attracted attention due to their number of applications in various industries viz. pharmaceuticals, food and bioethanol. Enzymes due to their unique properties and enormous power of catalysis at very wide range of temperature are always in demand in industries. There are certain techniques which are employed to improve the productivity of enzymes as well as enhancing their catalytic activity. Modeling of structure of inulinase will provide an overview of the catalytic domain and help in improvising catalytic potential. In the present review we have discussed on the topics of different substrate specificity and statistical optimization methods for the improvement of inulinase production. Recovery of enzyme is a cost effective approach and crucial step in the industrial application of enzyme and can be achieved by different immobilizing techniques. Immobilized inulinases have been widely studied and applied in different bioreactor systems. Kluyveromyces, Aspergillus, Staphylococcus, Xanthomonas, and Pseudomonas are few high level of inulinase producing microorganisms and are commercially employed for production of certain important product. Since inulins are used as prebiotic, it has also great impact in the nutritional biology field. Inulinase in food industries and inulin as probiotic are also discussed.

Probiotic Properties of Lactobacillus plantarum RYPR1 from an Indigenous Fermented Beverage Raabadi.

Present study documents the potential probiotic Lactobacillus isolated from indigenous fermented beverage Raabadi, consumed during summers in Haryana and Rajasthan regions of India. A total of five Raabadi samples were collected aseptically and 54 isolates were purified using MRS medium. All the isolates were assessed for tolerance to low pH and bile salts. It was observed that out of 54 only 24 isolates could survive the simulated gastric conditions. These isolates were further evaluated in vitro for cell surface hydrophobicity, cell surface hydrophobicity, hypocholesteramic activity, anti-oxidative potential, BSH activity, antagonistic activity, and antibiotic resistance profile. In addition, the confirmation of phenol resistance was also done. On the basis of results obtained, the survival rate of isolates was noted and six isolates were finally selected for further studies. Among them Lactobacillus plantarum RYPR1 and RYPC7 showed good survival at pH 2 which shows good acid tolerance. Moreover, L. plantarum RYPR1 showed the highest hydrophobicity (79.13%) and represented the deconjugation of bile salts, which help in their adhesion to epithelial cells and colonization. Furthermore, RYPR1 also exhibited highest cholesterol reduction (59%) and subsequent analysis of results revealed that the above mentioned isolates further exhibit a good hypocholesterolemic effect and could be possibly used to prevent hypercholesterolemia. The present study divulges that L. plantarum RYPR1 has an excellent probiotic potential.

KAP Study on HIV/AIDS Among Antenatal Women Attending Central Referral Hospital of North East India.

BACKGROUND: This study was conducted to determine whether antenatal mothers in Sikkim have adequate knowledge about awareness, attitude, and preventive practices regarding HIV infection. METHODS: Cross-sectional study using structured questionnaire. 220 Antenatal mothers attending the outpatient department of Central Referral Hospital of Sikkim were taken for the study for a period of 1 year from April 2011 to April 2012. Questionnaire form filled by pregnant women during their first antenatal visit was the source of data for this study. Systematic sampling technique was used where every alternate pregnant women registering for ANC visit were voluntarily recruited into the study. RESULTS: 2.27 % (5) women had not heard about HIV. 84 % (38) women had the knowledge that HIV was related to STI, while 50 % (110) did not. Television was the best method of increasing the knowledge (48 %). 68 % (150) of the women were aware about mother-to-child transmission (MTCT) of HIV during antenatal period. Only 2.66 % (6) women knew that HIV can be transmitted to child through breast milk. 90 % (198) knew that HIV is spread by having unsafe sex, 48 % (106) women knew using condoms would protect against it. 69.4 % (153) women wanted partner testing, and 84 % (185) of women consented that all pregnant women should be tested for HIV. CONCLUSIONS: The current study revealed high levels of knowledge, positive attitude, and preventive practices regarding HIV; however, this population lacked knowledge about MTCT and its prevention.

Catalytic Interactions and Molecular Docking of Bile Salt Hydrolase (BSH) from L. plantarum RYPR1 and Its Prebiotic Utilization.

Prebiotics are the non-digestible carbohydrate, which passes through the small intestine into unmetabolized form, reaches the large intestine and undergoes fermentation by the colonic bacteria thus; prebiotics stimulate the growth of probiotic bacteria. Further, bile salt hydrolase (BSH) is an enzyme that catalyses the deconjugation of bile salt, so it has enormous potential toward utilizing such capability of Lactobacillus plantarum RYPR1 toward detoxifying through BSH enzyme activity. In the present study, six isolates of Lactobacillus were evaluated for the co-aggregation assay and the isolate Lactobacillus plantarum RYPR1 was further selected for studies of prebiotic utilization, catalytic interactions and molecular docking. The prebiotic utilization ability was assessed by using commercially available prebiotics lactulose, inulin, xylitol, raffinose, and oligofructose P95. The results obtained revealed that RYPR1 is able to utilize these probiotics, maximum with lactulose by showing an increase in viable cell count (7.33 ± 0.02 to 8.18 ± 0.08). In addition, the molecular docking of BSH from Lactobacillus plantarum RYPR1 was performed which revealed the binding energy -4.42 and 7.03 KJ/mol. This proves a considerably good interactions among BSH and its substrates like Taurocholic acid (-4.42 KJ/mol) and Glycocholic acid (-7.03 KJ/mol). These results from this study establishes that Lactobacillus plantarum RYPR1 possesses good probiotic effects so it could be used for such applications. Further, molecular dynamics simulations were used to analyze the dynamic stability of the of modeled protein to stabilize it for further protein ligand docking and it was observed that residues Asn12, Ile8, and Leu6 were interacting among BSH and its substrates, i.e., Taurocholic acid and Lys88 and Asp126 were interacting with Glycocholic acid. These residues were interacting when the docking was carried out with stabilized BSH protein structure, thus, these residues may have a vital role in stabilizing the binding of the ligands with the protein.