Heterologous expression, purification and single step efficient refolding of recombinant tissue plasminogen activator (Reteplase) from E. coli.

Reteplase (recombinant plasminogen activator, rPA) is a mutant non-glycosylated tissue-type plasminogen activator (tPA) containing 355 amino acids with longer half-life and promising thrombolytic activity than its original counterpart, full length tPA. In this study, we aimed to produce and optimize the purification process of recombinant tissue-type plasminogen activator (tPA) known as Reteplase (rPA). Reteplase cDNA synthesized from total mRNA isolated from human placenta was PCR amplified, cloned into a pET-28a(+) E. coli expression vector and expressed in Rosetta-gami 2 E. coli (NovagenⓇ) host. rPA was expressed as an inclusion body in E. coli and its biological activity was achieved after single step solubilization, purification and refolding. We exploited the strategy of Slow Refolding using Gradual Dialysis (SRGD) in which a refolding buffer containing glutathione oxidized (1 mM GSSG) and glutathione reduced (3 mM GSH) and pH 9.0 was used. Using the SRGD method, we were able to successfully obtain the protein in its active form. We obtained 4.26 mg of active refolded protein from a 50 mL culture that was scaled up in a bioreactor. The purity and homogeneity of rPA was evaluated by SDS-PAGE, Western blotting and mass spectrometry. Circular dichroism spectroscopy was conducted to evaluate the refolding and stability of the refolded rPA in comparison to reference standard rPA. The thrombolytic potential of rPA was assessed by fibrin plate assay and In Vitro clot lysis assay. The presented protocol offers a viable approach for enhancing both the yield and refolding efficiency of reteplase, potentially resulting in an increase in yield.

Electrode Montage Induced Changes in Air-Conducted Ocular Vestibular-Evoked Myogenic Potential.

OBJECTIVES: Stimulus and recording parameters are pivotal for shaping the ocular vestibular-evoked myogenic potential (oVEMP). In the last decade, several attempts were made to identify the optimum electrode placement site to improve the oVEMP responses. A vast majority of these found larger response amplitudes for alternate electrode montages like belly-tendon (BT), chin-referenced (CR), and/or sternum-referenced montages than the clinically used infra-orbital montage. However, no study has yet compared all alternate electrode montages in a simultaneous recording paradigm to eliminate other confounding factors. Also, no study has compared all of them for their test-retest reliability, waveform morphology, and signal-to-noise ratio. Therefore, the decision on which among these electrode montages is best suited for oVEMP acquisition remains opaque. The present study aimed to investigate the effects of various electrode montages on oVEMP's response parameters and to determine the test-retest reliability of each of these in clinically healthy individuals using a simultaneous recording paradigm. DESIGN: This study had a within-subject experimental design. Fifty-five young healthy adults (age range: 20-30 years) underwent contralateral oVEMP recording using infra-orbital, BT, chin-referenced, and sternum-referenced electrode montages simultaneously using a four-channel evoked potential system. RESULTS: BT montage had a significantly shorter latency, larger amplitude, higher signal-to-noise ratio, and better morphology than other alternate montages ( p < 0.008). Further, all electrode montages of the current study showed fair/moderate to excellent test-retest reliability. CONCLUSIONS: By virtue of producing significantly better response parameters than the other electrode montages, BT montage seems better suited to the recording of oVEMP than the known electrode montages thus far.

Effect of Electrode Montage on Frequency Tuning Properties of Air-Conducted Ocular Vestibular-Evoked Myogenic Potential.

OBJECTIVES: The use of a 500 Hz tone burst over other frequencies was adopted for the clinical recording of ocular vestibular-evoked myogenic potential (oVEMP) on the basis that this stimulus frequency produces larger response amplitudes (frequency tuning) than the other frequencies. However, the possibility of reflex contamination due to a spatially displaced reference electrode from the muscle of the response origin raises questions about using an infraorbital (IO) montage. Nonetheless, the belly-tendon (BT) montage, which places both the recording electrodes over the inferior oblique muscle, increases the chances of obtaining a response with greater contribution from the inferior oblique muscle. However, whether this response continues to show the frequency tuning to 500 Hz is not known. Therefore, the present study aimed to examine the frequency tuning of oVEMP using various electrode montages. DESIGN: Thirty-eight young adults underwent simultaneous oVEMP recording from IO, BT, chin-referenced, and sternum-referenced electrode montages in response to 250, 500, 750, 1000, 1500, 2000, 3000, and 4000 Hz tone bursts. RESULTS: The frequency tuning most often coincided with a 750-Hz tone burst irrespective of the montage, with the BT montage exhibiting significantly higher response rates and larger peak to peak amplitudes than other montages (p < 0.008). Further, there was a "substantial" agreement on frequency tuning between BT and IO montages. CONCLUSIONS: With better response rates and response amplitudes yet similar frequency tuning to the IO montage, the BT montage can be a better option for the clinical recording of oVEMP across frequencies.

Unveiling the Health Benefits of Prebiotics: A Comprehensive Review.

Prebiotics play a pivotal role in fostering probiotics, essential contributors to the creation and maintenance of a conducive environment for beneficial microbiota within the human gut. To qualify as a prebiotic, a substance must demonstrate resilience to stomach enzymes, acidic pH levels, and intestinal bacteria, remaining unabsorbed in the digestive system while remaining accessible to gut microflora. The integration of prebiotics and probiotics into our daily diet establishes a cornerstone for optimal health, a priority for health-conscious consumers emphasizing nutrition that supports a balanced gut flora. Prebiotics offer diverse biological functions in humans, exhibiting antiobesity, antimicrobial, anticancer, anti-inflammatory, antidiabetic, and cholesterol-lowering properties, along with preventing digestive disorders. Numerous dietary fibers possessing prebiotic attributes are inadvertently present in our diets, emphasizing the broader significance of prebiotics. It is crucial to recognize that, while all dietary fibers are prebiotics, not all prebiotics fall under the category of dietary fibers. The versatile applications of prebiotics extend across various industries, such as dairy, bakery, beverages, cosmetics, pharmaceuticals, and other food products. This comprehensive review provides insights into different prebiotics, encompassing their sources, chemical compositions, and applications within the food industry.

Chemically Transformed Ag2 Te Nanowires on Polyvinylidene Fluoride Membrane For Flexible Thermoelectric Applications.

Flexible thermoelectric devices of nanomaterials have shown a great potential for applications in wearable to remotely located electronics with desired shapes and geometries. Continuous powering up the low power flexible electronics is a major challenge. We are reporting a flexible thermoelectric module prepared from silver telluride (Ag2 Te) nanowires (NWs), which are chemically transformed from uniquely synthesized and scalable tellurium (Te) NWs. Conducting Ag2 Te NWs composites have shown an ultralow total thermal conductivity ~0.22 W/mK surpassing the bulk melt-grown Ag2 Te ~1.23 W/mK at ~300 K, which is attributed to the nanostructuring of the material. Flexible thermoelectric device consisting of 4 legs (n-type) of Ag2 Te NWs on polyvinylidene fluoride membrane displays a significant output voltage (Voc ) ~2.3 mV upon human touch and Voc ~18 mV at temperature gradient, ΔT ~50 K, which shows the importance of NWs based flexible thermoelectric devices to power up the low power wearable electronics.

Utility of Inter-Frequency Amplitude Ratio of Vestibular-Evoked Myogenic Potentials in Identifying Meniere's Disease: A Systematic Review and Meta-Analysis.

OBJECTIVES: A recently devised parameter of vestibular-evoked myogenic potential (VEMP) based on the principles of frequency tuning is the inter-frequency amplitude ratio (IFAR). It refers to the ratio of the amplitude of 1000 Hz tone burst evoked VEMP to 500 Hz evoked tone burst. A pathology like Meniere's disease changes the frequency response and alters the frequency tuning of the otolith organs. Because IFAR is based on the principle of frequency tuning of VEMP, it is likely to help identify Meniere's disease. Few studies in the last decade have investigated the utility of IFAR in identifying Meniere's disease. However, a systematic review and a meta-analysis on IFAR in Meniere's disease are lacking. The present study investigates whether the IFAR of VEMP helps identify Meniere's disease and differentiates it from healthy ears and other vestibular pathologies. DESIGN: The present study is a systematic review and a meta-analysis. The studies investigating the IFAR of cervical and ocular VEMPs in Meniere's disease, healthy controls, and other vestibular pathologies were searched across research databases such as PubMed, Science Direct, and Scopus. The search strategy was developed using the PICO (population, intervention, comparison, and outcomes) format, and Medical Subject Headings (MeSH) terms and Boolean operators were employed. The systematic review was performed using the Rayyan software, whereas the Review Manager software was used to carry out the meta-analysis. A total of 16,605 articles were retrieved from the databases. After the duplicate removal, 2472 articles remained. These were eliminated using title screening, abstract screening, and full-length inspections. A total of nine articles were found eligible for quality assessment and meta-analysis, and the New Castle-Ottawa Scale was used for quality assessment. After the data extraction, 24 six articles were found to have the desired data format for the meta-analysis. RESULTS: The results showed significantly higher IFAR in the affected ears of individuals in the Meniere's disease group than in the control group's unaffected ears. There was no significant difference between the unaffected ears of individuals in the Meniere's disease group and the ears of the control group. The only study on Meniere's disease and benign paroxysmal positional vertigo found significantly larger ocular VEMP IFAR in ears with Meniere's disease than in benign paroxysmal positional vertigo. CONCLUSIONS: This systematic review and meta-analysis found IFAR efficient in differentiating Meniere's disease from healthy controls. We also found an enhanced IFAR as a potential marker for Meniere's disease. However, more investigations are needed to confirm the utility of an enhanced IFAR value in the exclusive identification of Meniere's disease.

Usefulness of masseter vestibular evoked myogenic potentials in identifying brainstem dysfunction among individuals with multiple sclerosis.

OBJECTIVE: The present study aimed to investigate the findings of cervical, ocular and masseter vestibular evoked myogenic potentials (cVEMP, oVEMP and mVEMP) among Multiple sclerosis (MS) and correlate with clinical and MRI findings. DESIGN: Standard group comparison research design. STUDY SAMPLE: Individuals with relapsing-remitting MS (n = 45) and age-sex-matched controls (n = 45) were the participants. All of them underwent case history, neurological examination, cVEMP, oVEMP and mVEMP testing. MRI was obtained only for MS participants. RESULTS: Abnormal result on at least one vestibular evoked myogenic potential (VEMP) sub-type was evidenced in 95.56% of participants whereas, unilateral or bilateral abnormal result on all three VEMP sub-types was observed in 60% of participants. The mVEMP abnormality was higher (82.22%) than cVEMP (75.56%) and oVEMP (75.56%) abnormalities but the differences were not significant (p > 0.05). There was no significant association of VEMP abnormalities with the presence of the brainstem symptoms, the brainstem signs, or the MRI lesions (p > 0.05). In the MS group, 38% had normal brainstem MRI; however, mVEMP, cVEMP and oVEMP abnormalities were evidenced in 82.4%, 64.7% and 52.94%, respectively. CONCLUSIONS: Among the three VEMP sub-types, mVEMP appears to be of greater value in identifying silent brainstem dysfunction undetected by clinical and MRI findings in the MS population.

Genetic Enhancement for Biotic Stress Resistance in Basmati Rice through Marker-Assisted Backcross Breeding.

Pusa Basmati 1509 (PB1509) is one of the major foreign-exchange-earning varieties of Basmati rice; it is semi-dwarf and early maturing with exceptional cooking quality and strong aroma. However, it is highly susceptible to various biotic stresses including bacterial blight and blast. Therefore, bacterial blight resistance genes, namely, xa13 + Xa21 and Xa38, and fungal blast resistance genes Pi9 + Pib and Pita were incorporated into the genetic background of recurrent parent (RP) PB1509 using donor parents, namely, Pusa Basmati 1718 (PB1718), Pusa 1927 (P1927), Pusa 1929 (P1929) and Tetep, respectively. Foreground selection was carried out with respective gene-linked markers, stringent phenotypic selection for recurrent parent phenotype, early generation background selection with Simple sequence repeat (SSR) markers, and background analysis at advanced generations with Rice Pan Genome Array comprising 80K SNPs. This has led to the development of Near isogenic lines (NILs), namely, Pusa 3037, Pusa 3054, Pusa 3060 and Pusa 3066 carrying genes xa13 + Xa21, Xa38, Pi9 + Pib and Pita with genomic similarity of 98.25%, 98.92%, 97.38% and 97.69%, respectively, as compared to the RP. Based on GGE-biplot analysis, Pusa 3037-1-44-3-164-20-249-2 carrying xa13 + Xa21, Pusa 3054-2-47-7-166-24-261-3 carrying Xa38, Pusa 3060-3-55-17-157-4-124-1 carrying Pi9 + Pib, and Pusa 3066-4-56-20-159-8-174-1 carrying Pita were identified to be relatively stable and better-performing individuals in the tested environments. Intercrossing between the best BC3F1s has led to the generation of Pusa 3122 (xa13 + Xa21 + Xa38), Pusa 3124 (Xa38 + Pi9 + Pib) and Pusa 3123 (Pi9 + Pib + Pita) with agronomy, grain and cooking quality parameters at par with PB1509. Cultivation of such improved varieties will help farmers reduce the cost of cultivation with decreased pesticide use and improve productivity with ensured safety to consumers.

Identification of novel vaccine candidates in the whole-cell Aeromonas hydrophila biofilm vaccine through reverse vaccinology approach.

Biofilm vaccine has been recognised as one of the successful strategy to reduce the Aeromonas hydrophila infection in fish. But, the vaccine contains the protective and non-protective proteins, which may lead to show altered heterologous adaptive immunity response. Moreover, cross protection and effectiveness of previously developed biofilm vaccine was not tested against different geographical A. hydrophila isolates. Therefore, in the present study, whole-cell A. hydrophila biofilm vaccine was evaluated in rohu, vaccinated group showed increased antibody titer and protection against the different geographical A. hydrophila isolates namely KAH1 and AAH2 with 78.9% and 84.2% relative percentage survival, respectively. In addition, by using the immune sera of biofilm vaccinated group, a total of six protective proteins were detected using western blot assay. Further, the same proteins were identified by nano LC-MS/MS method, a total of fourteen candidate proteins showing the immunogenic property including highly expressed OMP's tolC, bamA, lamb, AH4AK4_2542, AHGSH82_029580 were identified as potential vaccine candidates. The STRING analysis revealed that, top candidate proteins identified may potentially interact with other intracellular proteins; involved in ribosomal and (tricarboxylic acid) TCA pathway. Importantly, all the selected vaccine candidate proteins contain the B-cell epitope region. Finally, the present study concludes that, whole-cell A. hydrophila biofilm vaccine able to protect the fish against the different geographical A. hydrophila isolates. Further, through reverse vaccinology approach, a total of fourteen proteins were identified as potential vaccine candidates against A. hydrophila pathogen.

Characterizing the impact of advancing age on 500 Hz tone-burst evoked ocular-vestibular evoked myogenic potentials.

PURPOSE: Ageing is process that brings about steady decline in most of the systems, and vestibular system is no different. Although the effects of advancing age on the functioning of the semicircular canals and saccule has been well explored, its impact on the utricle remains the least studied component. While some researchers have investigated the effect of advancing age on the utricular functioning by virtue of studying ocular-vestibular evoked myogenic potentials (oVEMP), the use of smaller sample sizes would hamper the generalization of their results. Therefore, the present study aimed at investigating the effect of advancing age on oVEMP using a substantially larger sample size than the predecessors. METHODS: A total of 480 individuals in the age range of 10-70 years were used as participants and were divided into six age groups, each one defined as a decade. Contralateral oVEMPs were elicited by 500-Hz tone-bursts and averaged over 200 sweeps to obtain the final waveform. RESULTS: There was a significant decline in the response rates, prolongation of latencies and reduction of peak-to-peak amplitude of oVEMP with increasing age after 50 years (p < 0.05). However, there was no significant difference among the age groups until 50 years of age on any of these parameters (p > 0.05). Further, there was no significant change in inter-aural amplitude difference ratio with increasing age (p > 0.05). CONCLUSION: The results of the study confirm significant impact of the ageing process on oVEMP after the age of 50 years and, therefore, age appropriate norms should be considered when using oVEMP for investigating utricular functioning in an individual, especially after the age of 50 years.

Coding of consonant-vowel transition in children with central auditory processing disorder: an electrophysiological study.

INTRODUCTION: Acoustic change complex (ACC) is an important tool to investigate the encoding of the acoustic property of speech signals in various populations. However, there is a limited number of research papers that have explored the usefulness of ACC as a tool to study the neural encoding of consonant-vowel (CV) transition in children with central auditory processing disorder (CAPD). Thus, the present study aims to investigate the utility of ACC as an objective tool to study the neural representation of consonant-vowel (CV) transition in children with CAPD. METHODS: Twenty children diagnosed having CAPD and 20 normal counterparts in the age range of 8-14 years were the participants. The ACC was acquired using naturally produced CV syllable /sa/ with a duration of 380 ms. RESULTS: Latency of N1' and P2' was found to be prolonged in children with CAPD compared to normal counterparts, whereas the amplitude of N1' and P2' did not show any significant difference. Scalp topography showed significantly different activation patterns for children with and without CAPD. CONCLUSION: Prolonged latencies of ACC indicated poor encoding of CV transition in children with CAPD. The difference in scalp topography might be because of the involvement of additional brain areas for the neural discrimination task in children with CAPD.

Effect of advancing age on inter-frequency amplitude ratio of ocular vestibular evoked myogenic potentials.

OBJECTIVE: The inter-frequency amplitude ratio (IFAR) is the ratio of the amplitude of 1000-Hz tone-burst evoked ocular vestibular evoked myogenic potential (oVEMP) to the 500 Hz tone-burst evoked oVEMP. Since IFAR is an amplitude-based parameter, and the amplitudes of oVEMP for different frequencies are differentially affected by ageing, IFAR could potentially be affected by the ageing process. Therefore, we aimed to examine the effects of ageing on the IFAR of oVEMPs. DESIGN: Multiple static groups comparison. STUDY SAMPLE: We recorded oVEMPs for 500 Hz and 1000 Hz tone-bursts from 270 healthy individuals, each included in one of the six age groups. RESULTS: The IFAR was significantly larger in the ≥60 years age group than the age groups up to 49 years. Furthermore, the IFAR in the 50-59 years age group was significantly larger than all the other age groups up to 39 years. There was a significant positive correlation between age and IFARs (p < 0.01). CONCLUSIONS: IFAR, a sensitive tool in the test battery for the diagnosis of Meniere's disease, is affected even in healthy individuals due to ageing; hence its cautious interpretation is recommended when evaluating adults >50 years of age who are suspected of having Meniere's disease.

Relative efficacy of veria and mastoidectomy techniques of cochlear implantation in preservation of sound-induced saccular responses.

OBJECTIVE: Cochlear implantation (CI) is a safe technique to give hearing sensation to a person with hearing impairment. The present study aimed to compare the two surgical approaches of CI, mastoidectomy and veria, for their effects on saccular function assessed using cervical vestibular-evoked myogenic potential (cVEMP). DESIGN: Multiple group time series design. STUDY SAMPLE: The study included 63 children (3-8 years old) who underwent CI using veria technique (n = 20) and mastoidectomy approach (n = 43). The 500-Hz tone-burst evoked cVEMP were recorded on three occasions- a day before CI surgery, a day after the device switch-on and 4 months after the switch-on. RESULTS: The post-implant results revealed the absence of cVEMP in nearly 40% of the participants. The amplitudes were significantly lower at the time of the switch-on and at the 4-months follow-up period (p < 0.05). Among the participants undergoing CI using mastoidectomy approach, amplitudes were significantly larger after surgery than those undergoing surgery using veria technique (p < 0.05). CONCLUSIONS: The saccular responses are better preserved with the mastoidectomy technique than the veria technique for CI surgery.

Isolation and Characterization of Bacteriophages from Inland Saline Aquaculture Environments to Control Vibrio parahaemolyticus Contamination in Shrimp.

Among the various bacterial pathogens associated with the aquaculture environment, Vibrio parahaemolyticus the important one from shrimp and human health aspects. Though having been around for several decades, phage-based control of bacterial pathogens (phage therapy) has recently re-emerged as an attractive alternative due to the availability of modern phage characterization tools and the global emergence of antibiotic-resistant bacteria. In the present study, a total of 12 V. parahaemolyticus specific phages were isolated from 264 water samples collected from inland saline shrimp culture farms. During the host range analysis against standard/field isolates of V. parahaemolyticus and other bacterial species, lytic activity was observed against 2.3-45.5% of tested V. parahaemolyticus isolates. No lytic activity was observed against other bacterial species. For genomic characterization, high-quality phage nucleic acid with concentrations ranging from 7.66 to 220 ng/µl was isolated from 9 phages. After digestion treatments with DNase, RNase and S1 nuclease, the nature of phage nucleic acid was determined as ssDNA and dsDNA for 7 and 2 phages respectively. During transmission electron microscopy analysis of phage V5, it was found to have a filamentous shape making it a member of the family Inoviridae. During efficacy study of phage against V. parahaemolyticus in shrimp, 78.1% reduction in bacterial counts was observed within 1 h of phage application. These results indicate the potential of phage therapy for the control of V. parahaemoyticus in shrimp. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12088-021-00934-6.

Characterization and genome sequencing of three Aeromonas hydrophila-specific phages, CF8, PS1, and PS2.

Aeromonas hydrophila is an important finfish pathogen, besides being an opportunistic human pathogen. In the present study, the genomes of three A. hydrophila-specific phages, CF8, PS1, and PS2, were isolated, characterized and sequenced. Transmission electron microscopy showed that all three phages had typical Myoviridae morphology. The linear dsDNA genomes of CF8, PS1, and PS2 were 238,150 bp, 237,367 bp, and 240,447 bp in length, with a GC content of 42.2%, 38.8%, and 38.8%, respectively. The low sequence similarity (67.6% - 69.8% identity with 27.0% - 29.0% query coverage) to other phage genomes in the NCBI database indicated the novel nature of the CF8, PS1, and PS2 genomes. A total of 244, 247, and 250 open reading frames (ORFs) were predicted in the CF8, PS1, and PS2 genome, respectively. During the annotation process, functional predictions were made for 28-31 ORFs, while the rest were classified as "hypothetical proteins" with yet unknown functions. Genes for tRNAs were also detected in all phage genomes. As all three phages in the present study had a very narrow host range with lytic activity against only one strain of A. hydrophila, these phages could be good candidates for phage typing applications. Moreover, the endolysin- and lytic-transglycosylase-encoding genes could be used for recombinant cloning and expression of anti-microbial proteins.

Correction to: Characterization and genome sequencing of three Aeromonas hydrophila-specific phages, CF8, PS1, and PS2.

Authors would like to correct the incorrect version.

Neural representation of consonant-vowel transition in individuals with cochlear hearing loss and auditory neuropathy spectrum disorder.

PURPOSE: Acoustic change complex (ACC) is an evoked potential recorded in response to subtle change(s) in the continuing stimuli. It is assumed that poor speech perception can be due to poor encoding of consonant-vowel (CV) transition in cochlear hearing loss (CHL) and auditory neuropathy spectrum disorder (ANSD). The present study aims to investigate the use of ACC as an objective tool to study neural representation of CV transition in individuals with ANSD, CHL, and normal hearing (NH). METHODS: The study consisted of three groups of population (NH, ANSD, and CHL) in the age range of 18-40 years. ACC was recorded for naturally produced CV stimulus /sa/ of 380 ms in duration, which consists consonant (150 ms) and vowel (230 ms) using Biologic Navigator pro 7.2.1. RESULTS: The result showed significantly prolonged latencies of ACC in individuals with ANSD compared to NH. The current study also showed significantly prolonged latency and significantly lower peak-to-peak amplitude in individuals with ANSD compared to CHL. The interesting finding of the present study was significantly better peak-to-peak amplitude for CHL compared to NH. Whereas, there was no significant difference between NH and CHL for latencies measures. CONCLUSION: Poor neural synchronization in individuals with ANSD could be the reason of poor neural representation of CV transition in present study. The outcome of the present study showed poor neural representation of CV transition in individuals with ANSD compared to CHL and NH. Current study also showed better encoding of CV transition in individuals with CHL compared to ANSD.

Shotgun metagenomics offers novel insights into taxonomic compositions, metabolic pathways and antibiotic resistance genes in fish gut microbiome.

Gut microbiota of freshwater carp (Labeo rohita) was investigated by shotgun metagenomics to understand its taxonomic composition and functional capabilities. With the presence of 36 phyla, 326 families and 985 genera, the fish gut microbiota was found to be quite diverse in nature. However, at the phylum level, more than three-fourths of gut microbes belonged to Proteobacteria. Very low prevalence of commonly used probiotic bacteria (Bacillus, Lactobacillus, Streptococcus, and Lactococcus) in fish gut suggested the need to search for alternative probiotics for aquaculture use. Biosynthesis pathways were found to be the most dominant (51%) followed by degradation (39%), energy metabolism (4%) and fermentation (2%). In conformity with herbivorous feeding habit of L. rohita, gut microbiome also had pathways for the degradation of cellulose, hemicellulose, chitin, pectin, starch, and other complex carbohydrates. High prevalence of Actinobacteria and antibiotic biosynthesis pathways in the fish gut microbiome indicated its potential for bioprospecting of potentially novel natural antibiotics. Fifty-one different types of antibiotic resistance genes (ARGs) belonging to 15 antimicrobial resistance (AMR) gene families and conferring resistance against 24 antibiotic types were detected in fish gut. Some of the ARGs for multi-drug resistance were also found to be located on sequences of plasmid origin. The presence of pathogenic bacteria and ARGs on plasmid sequences suggested the potential risk due to horizontal gene transfer in the confined gut environment. The role of ARGs in fish gut microbiome needs further investigations.

Is There a Safe Level for Recording Vestibular Evoked Myogenic Potential? Evidence From Cochlear and Hearing Function Tests.

OBJECTIVE: There is a growing concern among the scientific community about the possible detrimental effects of signal levels used for eliciting vestibular evoked myogenic potentials (VEMPs) on hearing. A few recent studies showed temporary reduction in amplitude of otoacoustic emissions (OAE) after VEMP administration. Nonetheless, these studies used higher stimulus levels (133 and 130 dB peak equivalent sound pressure level [pe SPL]) than the ones often used (120 to 125 dB pe SPL) for clinical recording of VEMP. Therefore, it is not known whether these lower levels also have similar detrimental impact on hearing function. Hence, the present study aimed at investigating the effect of 500 Hz tone burst presented at 125 dB pe SPL on hearing functions. DESIGN: True experimental design, with an experimental and a control group, was used in this study. The study included 60 individuals with normal auditory and vestibular system. Of them, 30 underwent unilateral VEMP recording (group I) while the remaining 30 did not undergo VEMP testing (group II). Selection of participants to the groups was random. Pre- and post-VEMP assessments included pure-tone audiometry (250 to 16,000 Hz), distortion product OAE, and subjective symptoms. To simulate the time taken for VEMP testing in group I, participants in group II underwent these tests twice with a gap of 15 minutes. RESULTS: No participant experienced any subjective symptom after VEMP testing. There was no significant interear and intergroup difference in pure-tone thresholds and distortion product OAE amplitude before and after VEMP recording (p > 0.05). Furthermore, the response rate of cervical VEMP was 100% at stimulus intensity of 125 dB pe SPL. CONCLUSIONS: Use of 500 Hz tone burst at 125 dB pe SPL does not cause any temporary or permanent changes in cochlear function and hearing, yet produces 100% response rate of cervical VEMP in normal-hearing young adults. Therefore, 125 dB pe SPL of 500 Hz tone burst is recommended as safe level for obtaining cervical VEMP without significantly losing out on its response rate, at least in normal-hearing young adults.

Inter-frequency amplitude ratio of oVEMP for differentiating Meniere's disease from BPPV: clinical validation using a double-blind approach.

OBJECTIVE: The study aimed at examining the usefulness of inter-frequency amplitude ratio (IFAR) of ocular vestibular evoked myogenic potential (oVEMP) in identifying Meniere's disease (MD) and differentiating it from benign paroxysmal positional vertigo (BPPV). DESIGN: A case-control design was used with a double blind approach. Phase 1 included 70 healthy individuals and 36 individuals each with MD and BPPV and Phase 2 included 20 individuals each with MD and BPPV. The age range of the participants in both phases was 15-50 years. All participants underwent oVEMP testing using 500 and 1000 Hz tone bursts and IFAR was obtained. RESULTS: The results in phase 1 revealed significantly higher IFARs in Meniere's disease than BPPV and healthy individuals (p < 0.001). An optimum criterion point of IFAR ≥1.11 for diagnosing MD was found which yielded 80% sensitivity and 98% specificity. The results in phase 2 demonstrated 85% correct identification of MD and 95% correct rejection of BPPV as non-MD. CONCLUSIONS: IFAR of oVEMP appears highly sensitive and specific parameter for identifying MD and a clinically valid tool for differentiating MD from BPPV.