Exploring Possible Diagnostic Precancerous Biomarkers for Oral Submucous Fibrosis: A Narrative Review.

Oral submucous fibrosis (OSF) stands as a progressive oral ailment, designated as a potentially malignant disorder. OSF has gained widespread recognition as a significant precursor to malignant transformation. In the pursuit of dependable, straightforward, and non-invasive diagnostic measures for the early detection of oral malignant progression, research has delved into potential diagnostic biomarkers of OSF. This comprehensive review delves into current investigations that explore the correlation between various biomarkers and OSF. The molecular biomarkers of OSF are categorized based on cytology and sampling methods. Moreover, this review encompasses pertinent studies detailing how these biomarkers are acquired and processed. Within this scope, we scrutinize four potential biomarkers that hold the promise of facilitating the development of diagnostic tools for detecting early-stage OSF.

Combined Effects of Temperature and Salinity on the Pharmacokinetics of Florfenicol in Nile Tilapia (Oreochromis niloticus) Reared in Brackish Water.

Prudent antimicrobial use requires knowledge of pharmacokinetics (PK) in a specific fish species which in turn depends on water temperature and salinity. Although the influence of each individual factor is known, the combined effect is less clear. The objective of the current study was to investigate the effect of temperature and salinity concurrently on the PK of florfenicol (FF) in Nile tilapia reared in brackish water. Twenty-eight fish were divided into four groups and kept at one of two temperatures (24 vs. 32°C) and two salinity levels (5 vs. 15 ppt). The FF was administered at a single dose of 15 mg/kg body weight via oral gavage. The serum concentrations were analyzed by HPLC method and the PK parameters were analyzed by a 2-compartmental model. The result revealed that at 32°C, the elimination half-lives (t1/2ß), time to reach the peak concentration (Tmax), area under the serum concentration-time curve (AUC), and mean residence time (MRT) were significantly decreased, while the clearance relative to bioavailability (CL/F) significantly increased compared to those at 24°C. The extents of these PK changes were similar at the two salinity levels. On the contrary, increasing the salinity from 5 to 15 ppt at a given temperature level produced no significant change in the PK behavior. Our finding indicated that only water temperature, but not salinity, is the major determinant factor governing the FF fate in the fish body.

Determination of Optimal Doses and Minimum Effective Concentrations of Tricaine Methanesulfonate, 2-Phenoxyethanol and Eugenol for Laboratory Managements in Nile Tilapia (Oreochromis niloticus).

Anesthetic agents are often used in fish experiments to reduce the stress and struggle and to improve animal welfare. The present study aimed to determine the optimal doses and serum minimum effective concentration (MEC) of tricaine methanesulfonate (MS-222), 2-phenoxyethanol (2-PE), and eugenol (EUG) in Nile tilapia. Twenty-one fish were immersed in three different doses of each anesthetic and the minimal dose that produce stage III anesthesia within 5 min, maintain anesthesia status for 3 min, and recover within 5 min was considered the optimal dose. The serum concentrations of anesthetics immediately after the fish reached stage III anesthesia was defined as the MEC. The results revealed that the anesthetics dose-dependently shorten the induction time while the effect of doses on the recovery times were variable. The determined optimal doses for MS-222, 2-PE, and EUG were 300, 900, and 90 ppm, respectively. The MECs were 70, 263, and 53 µg/mL, respectively, about two to four times lower than the optimal doses and were independent of the doses. After immersion stopped, the serum concentrations decreased by >90% within the first hour and >99% after 4 h. Our research provides useful information for a smooth fish handling and design for researches requiring stage III anesthesia.

Postoperative Vomiting Following Laparoscopic Cholecystectomy Is Associated with Intraoperative Fluid Administration: A Retrospective Cohort Study.

Potential risk factors for postoperative vomiting (POV) are important for daily anesthesia practice. To identify the risk factors associated with POV we retrospectively reviewed 553 adult patients who underwent scheduled simple laparoscopic cholecystectomy under sevoflurane-based general anesthesia between January and December 2018. Patients who experienced POV were predominantly women, had lower body weight, and higher ASA (American Society of Anesthesiologists) physical status. The POV group showed female sex predominance, lower body weight, and higher ASA physical status, with a significant difference when compared with the non-POV group. In univariate analysis, female sex and Apfel scores of 2, 3, and 4 were associated with a higher POV incidence. Age > 70 years, higher body weight, and ASA physical status III were associated with a lower POV incidence. In multivariate logistic regression, sex, age, Apfel score, and intraoperative crystalloid infusion rate were POV predictive factors. Receiver operating characteristic analysis showed a negative association between the intraoperative crystalloid infusion rate and POV occurrence with an area under the curve of 0.73 (p = 0.001). The cutoff intraoperative crystalloid infusion rate was 2 mL/kg/h with 82% sensitivity and 49% specificity (≥2 mL/kg/h was associated with a lower POV incidence vs. <2 mL/kg/h (OR, 95% CI; 0.52 [0.33-0.83])). To decrease POV in these patients, identifying high-risk factors and an intraoperative crystalloid administration of ≥2 mL/kg/h should be considered in patients undergoing LC under sevoflurane-based general anesthesia.

Bath immersion pharmacokinetics of florfenicol in Nile tilapia (Oreochromis niloticus).

Drug administration by immersion can be a preferable method in certain conditions especially for treating small-sized, anorexic, or valuable fish. Pharmacokinetic information regarding bath treatment is considerably lacking in comparison to other common administration routes. The current study aimed to investigate if immersion can be an effective route to administer florfenicol (FF) for treatment in Nile tilapia. Nile tilapia reared at 28°C were immersed with FF solution at concentrations of 50, 100, 200, 500, and 500/200 (3 hr/117 hr) ppm for 120 hr and moved to drug-free freshwater for another 24 hr. The serum FF concentration in 100, 200, and 500/200 ppm groups reached steady-state at 12 hr with concentrations of 2.44, 3.04, and 5.26 µg/ml, respectively, which were about 2% of the bathing concentrations. The target therapeutic levels of 1-4 µg/ml were attained and maintained within 1-12 hr, depending on the immersion concentration and the target MIC. Serum FF reached the target with shorter time at higher bathing concentration. Following the 120-hr bath, the serum FF declined with the first-order half-life of approximately 10 hr. A minimum of 100 ppm FF is required for treatment purpose, and an initial high loading concentration followed by maintenance concentration is a plausible way to reach in vivo therapeutic level in short time. Greater than 99% of the residual FF in the bathing water could be removed within 15 min by 0.05% NaOCl. Our results indicated that bath immersion is a promising potential route for FF administration in Nile tilapia.

Pharmacokinetic-pharmacodynamic modelling for the determination of optimal dosing regimen of florfenicol in Nile tilapia (Oreochromis niloticus) at different water temperatures and antimicrobial susceptibility levels.

Optimized dosing regimen is key to the effective use of antibacterials and to minimizing drug-related side effects. The current study established a pharmacokinetic-pharmacodynamic (PK-PD) model for the determination of optimal antibacterial dosing regimen in fish taken into consideration the temperature-dependent PK and the pathogen-dependent antimicrobial susceptibility, using florfenicol (FF) in Nile tilapia as an example. The calculated optimal dosages significantly varied by temperature and target MIC levels, ranging from 2.23 (MIC 1 µg/ml at 24°C) to 34.88 mg kg-1  day-1 (MIC 4 µg/ml at 32°C). The appropriateness of the calculated dosages was successfully verified by the in vivo studies. After 5 days of oral administration of the calculated optimal dosage at 24°C, the predicted plasma drug values were in line with the mean observed Cmin(ss) while at 28 and 32°C underestimation of the Cmin(ss) in a dose-dependent manner was observed and likely due to the occurrence of non-linear PK at high dosages. The averaged serum protein binding of FF was 19.1%. Our results demonstrated the appropriateness and clinical applicability of the developed PK-PD approach for the determination of optimal dosing regimens at given temperatures and MICs. Saturation metabolism and PK non-linearity of FF in tilapia warrant further study.

Crayfish plague Aphanomyces astaci detected in redclaw crayfish, Cherax quadricarinatus in Taiwan.

Between December 2013 and January 2014, five outbreaks of an unknown disease with moderate to high cumulative mortality were observed among the freshwater redclaw crayfish (Cherax quadricarinatus) populations at four crayfish farms in Miaoli and Changhua counties (northern Taiwan) and at one crayfish farm in Pingtung County (southern Taiwan). Polymerase chain reaction (PCR) analysis allowed the detection of Aphanomyces astaci DNA in dead crayfish. Histopathological examination revealed an infection of host tissue by fungal hyphae that presented as typical non-septate hyphae within the soft abdominal cuticle from the first to second segment and in the tail fan. In PCR assays completed for the detection of crayfish plague, an expected 568-bp product, specific for the A. astaci ITS gene, was obtained from all sub-adults and adults examined. In a comparison of our strains with the known strains of A. astaci in Europe, nucleotide sequence identities were very similar, with 99.8-100% sequence similarity in that gene region. Positive reactions to in situ hybridization, using a digoxigenin (DIG)-labelled DNA probe, further confirmed A. astaci as the causative agent. This is the first report concerning natural infection of A. astaci in freshwater redclaw crayfish in Asia.

Effects of light-dark cycle on hippocampal iNOS expression and CREB activation in rats.

Nitric oxide (NO) is one of the most important interneural signaling molecules mediating hippocampal functions for learning and memory. The diurnal expression of neuronal NO synthase (nNOS) has been well studied. However, the temporal profile and underlying mechanisms of inducible NOS (iNOS) expression in a normal photoperiod or in altered light-dark cycles remain unclear. We examined the temporal profile of iNOS expression in adult male Sprague-Dawley rats maintained in a 12h-light/ 12h-dark photoperiod (12L/12D), which were pre-synchronized for 7 days before the experiment. The protein and mRNA levels of iNOS in the cortex and hippocampus were measured to examine the photic influences on iNOS expression. The results showed rhythmically changes of the levels of iNOS mRNA and protein in the hippocampus, but not in the cerebral cortex. The iNOS mRNA levels peaked at Zeitgeber time (ZT) 6 and ZT22, and the protein levels peaked at ZT8 and ZT18. Notably, the peaks in iNOS mRNA and protein levels in the 12L/12D group were 10 to 12 h apart, and the rhythmic pattern was absent in the 24-h period of the darkness group. In addition, the level of the phosphorylated cAMP response element binding protein (phospho-CREB) was the highest at ZT18, prior to a peak in iNOS mRNA expression at ZT22. A phospho-CREB-iNOS signaling pathway was further confirmed by the interaction of phospho-CREB and the iNOS DNA in histone complexes isolated by chromatin immunoprecipitation at ZT18. In conclusion, the photoperiod affects the diurnal expression of iNOS and activation of CREB in the hippocampus, which provide clues for the possible impact of circadian changes in hippocampal functions.

Memory consolidation and inducible nitric oxide synthase expression during different sleep stages in Parkinson disease.

BACKGROUND: Parkinson disease (PD) is a neurodegenerative disease characterized by motor and nonmotor dysfunctions, which include sleep disturbances. Rapid eye movement (REM) sleep is associated with numerous physiologic changes such as memory consolidation. Compelling evidence suggests that nitric oxide (NO) is crucial to both sleep regulation and memory consolidation. In our study, we explored changes in biologic molecules during various sleep stages and the effects of sleep on memory consolidation in PD. METHODS: Ten PD patients and 14 volunteers without PD participated in our study. The gene expression of inducible NO synthase (iNOS) in all sleep stages was measured using realtime polymerase chain reaction (PCR) based on polysomnography (PSG)-guided peripheral blood sampling. In addition, the efficiency of memory consolidation during the sleep of the participants was measured using the Wechsler Memory Scale, third edition (WMS-III). RESULTS: The iNOS expression increased in all sleep stages among the PD patients compared to the control participants, in whom iNOS expression decreased during REM sleep. Regarding memory consolidation, the performance of the controls in logic memory and the patients in visual reproduction tasks improved after sleep. CONCLUSIONS: The iNOS synthase expression was different from control participants among PD patients, and the expression was dissimilar in various sleep stages. Sleep might enhance memory consolidation and there are different memory consolidation profiles between PD and control participants demonstrating distinct memory consolidation profiles.

Production of monoclonal antibody against ORF72 of koi herpesvirus isolated in Taiwan.

A monoclonal antibody (MAb) was generated against the capsid protein (ORF 72) of koi herpesvirus (KHV) isolated from diseased koi Cyprinus carpio in Taiwan. The clone of MAb-B2 was obtained by immunizing mice with whole virus particles and further identified using indirect enzyme-linked immunosorbent assay and Western blot assay. In addition, it detected KHV in KHV-infected cells but not in those of mock-infected cells as demonstrated by indirect immunofluorescence assay. The neutralization test showed that MAb-B2 neutralized KHV. Furthermore, we uncovered that MAb-B2 recognizes the ORF72 of KHV as revealed by liquid chromatography-tandem mass spectrometry and Western blot assays. Additionally, MAb-B2 has been used as a diagnostic tool for detection of KHV in clinical samples by immunohistochemistry. Collectively, our results indicated that MAb-B2 could be used in the development of a diagnostic kit for diagnosis of KHV infections and ORF72 protein of KHV might be a candidate for future vaccine development.

Effects of C-terminal amino acids truncation on enzyme properties of Aeromonas caviae D1 chitinase.

C-terminal truncation mutagenesis was used to explore the functional and structural significance of the C-terminal region of Aeromonas caviae D1 chitinase (AcD1ChiA). Comparative studies between the engineered full-length AcD1ChiA and the truncated mutant (AcD1ChiAK606) included initial rate kinetics, fluorescence and circular dichroism (CD) spectrometric properties, and substrate binding and hydrolysis abilities. The overall catalytic efficiency, k(cat)/K(M), of AcD1ChiAK606 with the 4MU-(GlcNAc)(2) and the 4MU-(GlcNAc)(3) chitin substrates was 15-26% decreased. When compared with AcD1ChiA, the truncated mutant AcD1ChiAK606 maintained 80% relative substrate-binding ability and about 76% of the hydrolyzing efficiency against the insoluble alpha-chitin substrate. Both fluorescence and CD spectroscopy indicated that AcD1ChiAK606 retained the same conformation as AcD1ChiA. These results indicated that removal of the C-terminal 259 amino acid residues, including the putative chitin-binding motif and the A region (a motif of unknown function) of AcD1ChiA, did not seriously affect the enzyme structure integrity as well as activity. The present study provided evidences illustrating that the binding and hydrolyzing of insoluble chitin substrates by AcD1ChiA were not absolutely dependent on the putative C-terminal chitin-binding domain and the function-unknown A region.

Phylogenetic characterization of Newcastle disease viruses isolated in Taiwan during 2003-2006.

Isolates of Newcastle disease virus (NDV) from chicken cases were obtained from various locations in Taiwan during 2003-2006 and were genotypically analyzed by using reverse transcription polymerase chain reaction (RT-PCR) with primers specific to the viral fusion (F) protein gene (534 bp). Part of the amplified F protein DNA product (nucleotide sequence 47-418) and the deduced amino acid sequences were compared phylogenetically with those from strains previously reported in Taiwan and other geographic regions. Our results showed that all Taiwanese isolates (n=20) collected during 2003-2006, according to the phylogenetic tree, belong to the genotype VIId. In addition, all the six Taiwanese isolates obtained in 2003, carry the motif (112)R-R-Q-K-R(116) and have the amino acid L(23) replaced by F(23) (assigned as Group 1). On the other hand, 12 out of the 14 Taiwanese isolates obtained during 2004-2006 possess the motif (112)R-R-K-K-R(116) and have the amino acid G(74), instead of E(74) (assigned as Group 2). To our best knowledge, this is the first reported VIId isolates that possess the sequences of G(74)/(112)R-R-K-K-R(116) within the F0 protein. Since a high mortality, severe clinical signs, typical postmortem lesions, and a high intra-cerebral pathogenicity index (ICPI) were observed in the NDV-infected chickens, these isolates acquired between 2003 and 2006 are considered as the velogenic type. The Group 2 viruses have become dominant and responsible for the majority of Taiwanese outbreaks during recent years. Based on our phylogenetic analysis, it can be postulated that these isolates were evolved from previously reported local strains, and the Group 2 family emerged the latest in the genotype VIId. The information is fundamental to improving the efficiency of controlling strategies and vaccine development for NDV.

PCR and in situ hybridization for the detection and localization of a new pathogen Francisella-like bacterium (FLB) in ornamental cichlids.

Archived formalin-fixed, paraffin-embedded tissues from 28 diseased ornamental cichlid fish associated with visceral granulomas were examined by polymerase chain reaction (PCR) and in situ hybridization (ISH) for detection of Francisella-like bacteria (FLB). The 16S rDNA FLB-specific primer pair 180f/465r was used on naturally infected ornamental cichlids, resulting in 11 positive cases (39%). Using DNA probes, all 28 cases (100%) showed a positive reaction, and most labeled cells were observed in the visceral granulomas of infected individuals. FLB was detected in cells morphologically resembling epithelioid and endothelioid macrophages. ISH was more sensitive than PCR or routine histopathological examination, based on the examination of archived formalin-fixed, paraffin-embedded tissues in this study. Furthermore, this technique located a new fish pathogen, FLB, in ornamental cichlids. The causative agent was similar to the pathogen inducing systemic granulomas in tilapia.

Pathological and molecular studies on mycobacteriosis of milkfish Chanos chanos in Taiwan.

investigated in milkfish Chanos chanos, which had a cumulative mortality of up to 66.7% over the course of 1 yr. Gross reddish- or greyish-white nodules appeared on the peritoneal surface, spleen, kidney, liver and gastrointestinal (GI) tract. Epithelioid granulomas with the formation of Langhan's type giant cells were the prominent histopathological changes. Despite large numbers of acid-fast bacilli in the granulomas, neither caseous necrosis nor dystrophic calcification were observed. Using degenerate primers that targeted the heat shock protein 65 kDa gene of Mycobacterium spp., a 441 bp product was amplified. When compared with published sequences, our products were identical to those of Mycobacterium abscessus Type II (GenBank accession number AY603554). This is the first report of M. abscessus infection in milkfish.