Gonad recrudescence and annual sex steroid patterns in female goldspot mullet, Liza parsia reared in brackishwater pond.

Goldspot mullet, Liza parsia is a commercially important fish of South East Asia, where its farming depends on wild seed resources due to unavailability of hatchery technology. It, therefore, is important to understand the annual reproductive cycle of female L. parsia in captivity. In this study, adult male and female L. parsia (body weight ranges: 45-90 g; total length ranges: 100-125 mm, age >1 year) were collected from the wild and reared in a brackishwater pond. Thereafter, fish were randomly sampled at monthly intervals to measure the hepatosomatic index (HSI), gonadosomatic index (GSI), levels of serum steroids (testosterone, T; 17ß-estradiol, E2 and 17α-hydroxyprogesterone, 17-OHP), and oocyte growth. Results exhibited that female L. parsia undergoes six different maturation stages, namely I (oocyte diameter, OD: <100 µm), II (OD: 100-350 µm), III (OD: 350-400 µm), IV (vitellogenic oocyte, OD: 400-450 µm), V (ripe oocyte, OD: 450-550 µm) and VI (atretic oocyte, OD: 60-150 µm), with synchronous oocyte development. The highest (P < 0.05) HSI (1.96 ± 0.24) and GSI (12.01 ± 0.73) were recorded in December and January, respectively. Concentration of E2 gradually increased from August and reached its peak (807.67 ± 25.98 pg mL-1, P < 0.05) in December. The level of 17-OHP (85.87 ± 0.91 pg mL-1) was at its peak during the normal spawning month (January) (P < 0.05). Overall, the results indicated that L. parsia attains maturity in brackishwater pond, which is consistent with previous observations, and altogether provide the basis to develop a breeding technology in captivity through hormonal and environmental manipulations.

Exploration of binding mechanism of triclosan towards cancer markers using molecular docking and molecular dynamics.

The molecule 5-chloro-2-(2,4-dichlorophenoxy) phenol is well-known as Triclosan (TCS), which is also a potential endocrine disrupting synthetic chemical. TCS exposure has been connected to the control of the human enoyl-acyl carrier protein-reductase (hER), which has been linked to a range of life threatening diseases. However, other than hER, the new protein targets for TCS that are responsible for a variety of cancers are yet unclear. The goal of this work is to investigate into the protein binding patterns of TCS and proteins from various cancer signaling pathways. Discovery Studio 4.1 was used to perform molecular docking and molecular dynamics (MD) on the protein-triclosan complex. The proteins were first screened using CHARMM-based docking with a CDOCKER energy greater than -21.40 kcal/mol. The CDOCKER energies of Fas-associated death domain (FADD), Receptor-interacting protein 1 (RIP1), F-κB-inducing kinase (NIK), c-Jun N-terminal kinase (JNK), Apoptosis signal-regulating kinase 1 (ASK1), B-cell lymphoma 2 (Bcl-2), Apoptosis-inducing factor (AIF), α-tubulin, and Actin were -20.68 kcal/mol, -26.88 kcal/mol, -23.43 kcal/mol, -22.21 kcal/mol, -20.40 kcal/mol, -21.10 kcal/mol, -20.98 kcal/mol, -24.67 kcal/mol, and -23.09 kcal/mol respectively. MD was performed on the screened proteins by standard dynamics cascade tool using CHARMM Force field. The MD results were accessed using the energy-time graph, root-mean-square deviation (RMSD), and root mean square fluctuations (RMSF). The 100 conformers of α-tubulin, NIK, FADD, and RIP1 were found to have a trend of increasing RMSD, whereas Bcl-2, ASK1, AIF, Actin, and JNK proteins had lower RMSD values. In compared to FADD, AIF, and JNK, the RMSF variations of the Bcl-2, ASK1, α-tubulin, Actin, NIK, and RIP1 residues were shown to be high. Similar patterns were seen in the energy variations, which range from 1000 kcal/mol to 2000 kcal/mol. RIP1 and Bcl-2 showed more variation in the sidechain RMSF in comparison to FADD, ASK1, AIF, Actin, α-tubulin, NIK and JNK. Thus, it can be postulated that AIF and JNK proteins of apoptosis signaling pathway are pivotal in the TCS mediated reactions.

Identifying research questions for HIV, tuberculosis, tuberculosis-HIV, malaria, and neglected tropical diseases through the World Health Organization guideline development process: a retrospective analysis, 2008-2018.

OBJECTIVES: World Health Organization (WHO) guidelines for health programmes and healthcare delivery are the foundation of its technical leadership in public health and essential to decision-making globally. A key function of guideline development is to identify areas in which further evidence is needed because filling these gaps will lead to future improvements in population health. The objective of this study was to examine the knowledge gaps and research questions for addressing those gaps generated through the WHO guideline development process, with the goal of informing future strategies for improving and strengthening the guideline development process. STUDY DESIGN: We did a systematic, retrospective analysis of research questions identified in the published guidelines. METHODS: We analyzed guidelines published between January 1, 2008, and December 31, 2018, by the Communicable Diseases Cluster in five disease areas: tuberculosis (TB), HIV, malaria, TB-HIV, and neglected tropical diseases (NTDs). Research questions were extracted independently by two researchers. We analyzed the distribution of research questions by disease and by topic category and did a qualitative assessment of optimum practice for research question generation during the guideline development process. RESULTS: A total of 48 guidelines were included: 26 on HIV, 1 on malaria, 11 on TB, 5 on TB/HIV, and 5 on NTDs. Overall, 36 (75%) guidelines encompassed a total of 360 explicit research questions; the remainder did not contain specific research questions. The number of research questions that focused on TB was 49, TB/HIV was 38, HIV was 250, and NTDs was 23. The number of research questions that focused on diagnosis was 43 (11.9%) of 360, prevention was 62 (17.2%), treatment was 103 (28.6%), good practice was 12 (3.3%), service delivery was 86 (23.8%), and other areas was 54 (15%). Research questions were often not formulated in a specific or actionable way and were hard to identify in the guideline. Examples of good practice identified by the review team involved the generation of specific and narrowly defined research questions, with accompanying recommendations for appropriate study design. CONCLUSIONS: The WHO must strengthen its approach to identifying and presenting research questions during the guideline development process. Ensuring access to research questions is a key next step in adding value to the guideline development process.

Docking-based inverse virtual screening strategy for identification of novel protein targets for triclosan.

Triclosan (TCS) is chemically designated as 5-chloro-2-(2,4-dichlorophenoxy) phenol and is considered as endocrine-disrupting chemical (EDC). The various diseases found due to exposure of TCS, have been linked with modulation of the human enoyl-acyl carrier protein-reductase (hER). However, the new protein targets for TCS other than hER, which are responsible for various diseases, are still unknown. In the present study, a bioinformatics approach was used to identify new possible targets for TCS. A text mining study was initially performed to understand the association of TCS in various biochemical processes. Discovery studio software 4.1 was used to carry out inverse virtual screening for 226 numbers of pathway proteins by docking study using CHARMm based docking tool, and twenty proteins were screened. CDOCKER energy values lower than -12.65 kcal/mol was considered for the screening of selected proteins. Three new proteins; Receptor-interacting protein 1 (RIP1), Apoptosis signal-regulating kinase 1 (ASK1) and B-cell lymphoma 2 (Bcl-2) from Apoptosis Signaling Pathway revealed best CDOCKER energy with triclosan which was -26.88, -23.34 and -22.96 kcal/mol respectively. The interaction of TCS with RIP1 and ASK1 were mostly hydrophobic; however, hydrogen bond type interaction was found in TCS/Bcl2 complex. Therefore, docking-based inverse virtual screening study suggests that TCS has other targets rather than hER, which can modulate various biochemical processes. The docking protocol was validated through evaluation of root-mean-square deviation (RMSD), key interaction score system (KISS) and the relationship between the docking energy and toxicity data available in ToxCast database. Low RMSD value (0.55 ˚A) and high KISS score (0.66) along with higher correlation (R2 = 0.9798) between docking affinity and toxicity indicated that docking protocol can be used to optimize the binding energetics.

A Standard Mutual Authentication Protocol for Cloud Computing Based Health Care System.

Telecare Medical Information System (TMIS) supports a standard platform to the patient for getting necessary medical treatment from the doctor(s) via Internet communication. Security protection is important for medical records (data) of the patients because of very sensitive information. Besides, patient anonymity is another most important property, which must be protected. Most recently, Chiou et al. suggested an authentication protocol for TMIS by utilizing the concept of cloud environment. They claimed that their protocol is patient anonymous and well security protected. We reviewed their protocol and found that it is completely insecure against patient anonymity. Further, the same protocol is not protected against mobile device stolen attack. In order to improve security level and complexity, we design a light weight authentication protocol for the same environment. Our security analysis ensures resilience of all possible security attacks. The performance of our protocol is relatively standard in comparison with the related previous research.

An Efficient and Practical Smart Card Based Anonymity Preserving User Authentication Scheme for TMIS using Elliptic Curve Cryptography.

In the last few years, numerous remote user authentication and session key agreement schemes have been put forwarded for Telecare Medical Information System, where the patient and medical server exchange medical information using Internet. We have found that most of the schemes are not usable for practical applications due to known security weaknesses. It is also worth to note that unrestricted number of patients login to the single medical server across the globe. Therefore, the computation and maintenance overhead would be high and the server may fail to provide services. In this article, we have designed a medical system architecture and a standard mutual authentication scheme for single medical server, where the patient can securely exchange medical data with the doctor(s) via trusted central medical server over any insecure network. We then explored the security of the scheme with its resilience to attacks. Moreover, we formally validated the proposed scheme through the simulation using Automated Validation of Internet Security Schemes and Applications software whose outcomes confirm that the scheme is protected against active and passive attacks. The performance comparison demonstrated that the proposed scheme has lower communication cost than the existing schemes in literature. In addition, the computation cost of the proposed scheme is nearly equal to the exiting schemes. The proposed scheme not only efficient in terms of different security attacks, but it also provides an efficient login, mutual authentication, session key agreement and verification and password update phases along with password recovery.

Cryptanalysis and Enhancement of Anonymity Preserving Remote User Mutual Authentication and Session Key Agreement Scheme for E-Health Care Systems.

The E-health care systems employ IT infrastructure for maximizing health care resources utilization as well as providing flexible opportunities to the remote patient. Therefore, transmission of medical data over any public networks is necessary in health care system. Note that patient authentication including secure data transmission in e-health care system is critical issue. Although several user authentication schemes for accessing remote services are available, their security analysis show that none of them are free from relevant security attacks. We reviewed Das et al.'s scheme and demonstrated their scheme lacks proper protection against several security attacks such as user anonymity, off-line password guessing attack, smart card theft attack, user impersonation attack, server impersonation attack, session key discloser attack. In order to overcome the mentioned security pitfalls, this paper proposes an anonymity preserving remote patient authentication scheme usable in E-health care systems. We then validated the security of the proposed scheme using BAN logic that ensures secure mutual authentication and session key agreement. We also presented the experimental results of the proposed scheme using AVISPA software and the results ensure that our scheme is secure under OFMC and CL-AtSe models. Moreover, resilience of relevant security attacks has been proved through both formal and informal security analysis. The performance analysis and comparison with other schemes are also made, and it has been found that the proposed scheme overcomes the security drawbacks of the Das et al.'s scheme and additionally achieves extra security requirements.

Design and Analysis of an Enhanced Patient-Server Mutual Authentication Protocol for Telecare Medical Information System.

In order to access remote medical server, generally the patients utilize smart card to login to the server. It has been observed that most of the user (patient) authentication protocols suffer from smart card stolen attack that means the attacker can mount several common attacks after extracting smart card information. Recently, Lu et al.'s proposes a session key agreement protocol between the patient and remote medical server and claims that the same protocol is secure against relevant security attacks. However, this paper presents several security attacks on Lu et al.'s protocol such as identity trace attack, new smart card issue attack, patient impersonation attack and medical server impersonation attack. In order to fix the mentioned security pitfalls including smart card stolen attack, this paper proposes an efficient remote mutual authentication protocol using smart card. We have then simulated the proposed protocol using widely-accepted AVISPA simulation tool whose results make certain that the same protocol is secure against active and passive attacks including replay and man-in-the-middle attacks. Moreover, the rigorous security analysis proves that the proposed protocol provides strong security protection on the relevant security attacks including smart card stolen attack. We compare the proposed scheme with several related schemes in terms of computation cost and communication cost as well as security functionalities. It has been observed that the proposed scheme is comparatively better than related existing schemes.

An Improved RSA Based User Authentication and Session Key Agreement Protocol Usable in TMIS.

Recently, Giri et al.'s proposed a RSA cryptosystem based remote user authentication scheme for telecare medical information system and claimed that the protocol is secure against all the relevant security attacks. However, we have scrutinized the Giri et al.'s protocol and pointed out that the protocol is not secure against off-line password guessing attack, privileged insider attack and also suffers from anonymity problem. Moreover, the extension of password guessing attack leads to more security weaknesses. Therefore, this protocol needs improvement in terms of security before implementing in real-life application. To fix the mentioned security pitfalls, this paper proposes an improved scheme over Giri et al.'s scheme, which preserves user anonymity property. We have then simulated the proposed protocol using widely-accepted AVISPA tool which ensures that the protocol is SAFE under OFMC and CL-AtSe models, that means the same protocol is secure against active and passive attacks including replay and man-in-the-middle attacks. The informal cryptanalysis has been also presented, which confirmed that the proposed protocol provides well security protection on the relevant security attacks. The performance analysis section compares the proposed protocol with other existing protocols in terms of security and it has been observed that the protocol provides more security and achieves additional functionalities such as user anonymity and session key verification.

A Secure Three-Factor User Authentication and Key Agreement Protocol for TMIS With User Anonymity.

Telecare medical information system (TMIS) makes an efficient and convenient connection between patient(s)/user(s) and doctor(s) over the insecure internet. Therefore, data security, privacy and user authentication are enormously important for accessing important medical data over insecure communication. Recently, many user authentication protocols for TMIS have been proposed in the literature and it has been observed that most of the protocols cannot achieve complete security requirements. In this paper, we have scrutinized two (Mishra et al., Xu et al.) remote user authentication protocols using smart card and explained that both the protocols are suffering against several security weaknesses. We have then presented three-factor user authentication and key agreement protocol usable for TMIS, which fix the security pitfalls of the above mentioned schemes. The informal cryptanalysis makes certain that the proposed protocol provides well security protection on the relevant security attacks. Furthermore, the simulator AVISPA tool confirms that the protocol is secure against active and passive attacks including replay and man-in-the-middle attacks. The security functionalities and performance comparison analysis confirm that our protocol not only provide strong protection on security attacks, but it also achieves better complexities along with efficient login and password change phase as well as session key verification property.

A novel user authentication and key agreement protocol for accessing multi-medical server usable in TMIS.

Telecare Medical Information System (TMIS) makes an efficient and convenient connection between patient(s)/user(s) at home and doctor(s) at a clinical center. To ensure secure connection between the two entities (patient(s)/user(s), doctor(s)), user authentication is enormously important for the medical server. In this regard, many authentication protocols have been proposed in the literature only for accessing single medical server. In order to fix the drawbacks of the single medical server, we have primarily developed a novel architecture for accessing several medical services of the multi-medical server, where a user can directly communicate with the doctor of the medical server securely. Thereafter, we have developed a smart card based user authentication and key agreement security protocol usable for TMIS system using cryptographic one-way hash function. We have analyzed the security of our proposed authentication scheme through both formal and informal security analysis. Furthermore, we have simulated the proposed scheme for the formal security verification using the widely-accepted AVISPA (Automated Validation of Internet Security Protocols and Applications) tool and showed that the scheme is secure against the replay and man-in-the-middle attacks. The informal security analysis is also presented which confirms that the protocol has well security protection on the relevant security attacks. The security and performance comparison analysis confirm that the proposed protocol not only provides security protection on the above mentioned attacks, but it also achieves better complexities along with efficient login and password change phase.

Protoplast-to-plant regeneration of American elm (Ulmus americana).

This study describes a protocol for regeneration of plants from cell suspension-derived protoplasts of American elm (Ulmus americana). Efficient protoplast isolation was achieved from a two-phase culture system through the incorporation of 100 µM 2-aminoindan-2-phosphonic acid, with a yield of approximately 2 × 10(6) protoplasts/ml packed cell volume. Isolated protoplasts failed to survive in liquid or alginate bead culture systems but initiated and continued to divide when embedded in low melting point agarose beads. Protoplast-derived callus proliferated and differentiated into shoot buds in response to 10 or 20 µM thidiazuron. Differentiated buds elongated and continued to proliferate on elm shoot medium supplemented with 3.0 µM GA3. The protoplast-derived shoots rooted and acclimatized to greenhouse conditions and continued to grow. This system provides the first protoplast-to-plant regeneration system for American elm and provides a framework for the development of protoplast fusion or genome editing technologies.

ERRß signalling through FST and BCAS2 inhibits cellular proliferation in breast cancer cells.

BACKGROUND: The overexpression of oestrogen-related receptor-ß (ERRß) in breast cancer patients is correlated with improved prognosis and longer relapse-free survival, and the level of ERRß mRNA is inversely correlated with the S-phase fraction of cells from breast cancer patients. METHODS: Chromatin immunoprecipitation (ChIP) cloning of ERRß transcriptional targets and gel supershift assays identified breast cancer amplified sequence 2 (BCAS2) and Follistatin (FST) as two important downstream genes that help to regulate tumourigenesis. Confocal microscopy, co-immunoprecipitation (CoIP), western blotting and quantitative real-time PCR confirmed the involvement of ERRß in oestrogen signalling. RESULTS: Overexpressed ERRß induced FST-mediated apoptosis in breast cancer cells, and E-cadherin expression was also enhanced through upregulation of FST. However, this anti-proliferative signalling function was challenged by ERRß-mediated BCAS2 upregulation, which inhibited FST transcription through the downregulation of ß-catenin/TCF4 recruitment to the FST promoter. Interestingly, ERRß-mediated upregulation of BCAS2 downregulated the major G1-S transition marker cyclin D1, despite the predictable oncogenic properties of BCAS2. INTERPRETATION: Our study provides the first evidence that ERRß, which is a coregulator of ERα also acts as a potential tumour-suppressor molecule in breast cancer. Our current report also provides novel insights into the entire cascade of ERRß signalling events, which may lead to BCAS2-mediated blockage of the G1/S transition and inhibition of the epithelial to mesenchymal transition through FST-mediated regulation of E-cadherin. Importantly, matrix metalloprotease 7, which is a classical mediator of metastasis and E-cadherin cleavage, was also restricted as a result of ERRß-mediated FST overexpression.

Primary soft tissue Ewing's sarcoma of the maxillary sinus in elderly patients: presentation, management and prognosis.

Nonosseous or soft tissue Ewing's sarcoma is a rare form of Ewing's sarcoma/primitive neuroectodermal tumour that seldom affects the head and neck region. Involvement of the nose and paranasal sinuses is extremely uncommon, with only eight of such patients being reported to date, mostly affecting adolescents and young adults. To our knowledge, this study is the first comprehensive report of primary soft tissue Ewing's sarcoma involving the paranasal sinuses in an elderly patient who successfully completed treatment. We herein discuss the pathogenesis, management and factors affecting the prognosis of this rare group of tumours involving the nose and paranasal sinuses, in relation to the available literature.

Isolation of lactic acid bacteria from kuruma shrimp (Marsupenaeus japonicus) intestine and assessment of immunomodulatory role of a selected strain as probiotic.

Fifty-one lactic acid bacteria (LAB) strains were isolated and identified based on 16S ribosomal DNA sequence from the intestinal tracts of 142 kuruma shrimps (Marsupenaeus japonicus) collected from Kanmon Strait, Fukuoka and Tachibana Bay, Nagasaki, Japan. Cellular immunomodulatory function of 51 isolated LAB strains was assessed by measuring the level of interferon (IFN)-γ induction in mouse spleen cell culture. The strain Lactococcus lactis D1813 exhibited the highest amount of IFN-γ production and also bactericidal activity and was selected for testing its immunomodulatory role as a probiotic in kuruma shrimp. We also assessed the effect of dietary incorporation of this probiotic on resistance to Vibrio penaeicida infection in the kuruma shrimp. Our results demonstrate that probiotic L. lactis D1813-containing diet-fed (105 cfu g⁻¹) shrimps displayed a significant up-regulation of lysozyme gene expressions in the intestine and hepatopancreas. However, insignificantly higher expression of anti-lipopolysaccharide factor, super oxide dismutase, prophenoloxidase, and toll-like receptor 1 was recorded in the intestine of shrimps fed the probiotic diet. Moreover, significantly increased (P < 0.01) resistance to the bacterial pathogen in term of better post-infection survival (61.7 %) was observed in the shrimps fed with the probiotic-incorporated diet compared with the control diet-fed group (28.3 %). The present study indicates the immunomodulatory role of the LAB L. lactis D1813 on the kuruma shrimp immune system and supports its potential use as an effective probiotic in shrimp aquaculture.

Cytokine mediated immune responses in the Japanese pufferfish (Takifugu rubripes) administered with heat-killed Lactobacillus paracasei spp. paracasei (06TCa22) isolated from the Mongolian dairy product.

The important role played by cytokines in host innate immunity and the interaction of subsets of immune and inflammatory cells through cytokines offer avenues for immune interventions. We investigated 16 cytokine gene responses in the Japanese pufferfish, Takifugu rubripes orally treated with a heat-killed lactic acid bacterium (LAB), Lactobacillus paracasei spp. paracasei (strain 06TCa22) (Lpp) isolated from a Mongolian dairy product at 1mgg(-1)body weightd(-1) for 3days. Additionally, we assessed superoxide anion production (SAP) and phagocytic activity (PA) of head kidney cells and resistance to Vibrio harveyi infection in treated fish. Significant up-regulation of pro-inflammatory (IL-1ß, IL-6, IL-17A/F-3, TNF-α and TNF-N), cell-mediated immunity inducing (IL-12p35, IL-12p40 and IL-18), antiviral/intra-cellular pathogen killing (I-IFN-1 and IFN-γ), anti-inflammatory (IL-10) and peripheral T cell expansion and survival controlling (IL-2, IL-7, IL-15, IL-21 and TGF-ß1) cytokines was observed in the treated fish. Furthermore, significantly increased SAP, PA and pathogen resistance were observed in the treated fish compared to untreated fish. Our results indicate the enhancement of cytokine mediated immunity in T. rubripes by the use of the heat-killed Lpp as a potential immunostimulant and would be of great use in immunomodulation trials with the possibility to monitor positive immune response.

Evaluation of the efficacy of a combination of diltiazem and periprostatic nerve block in pain control during transrectal ultrasonography-guided biopsy of the prostate.

INTRODUCTION: The choice of analgesia during prostate biopsy remains controversial. The pain has dual origin: from the insertion of the probe as well as the biopsy itself. Periprostatic nerve block (PPNB) is currently the gold standard modality for decreasing pain of prostate biopsy but it does not alleviate the pain of probe insertion. A randomised controlled trial was performed to test the efficacy and safety of the combination of topical application of diltiazem gel and PPNB for pain control during transrectal ultrasonography guided prostate biopsy. METHODS: A total of 73 patients who were to undergo their first prostate biopsy were randomised to receive either 2ml of 2% topical diltiazem gel or a placebo 15 minutes before the biopsy. All the patients then had a PPNB using 1% lignocaine. A ten- point visual analogue scale was used to record the pain immediately after the insertion of the probe and during the biopsy. Any adverse effects were also recorded. RESULTS: There was no significant difference in the mean age and prostate volumes between the groups. There was a significantly lower mean pain score due to probe insertion in those patients who received topical diltiazem than in the placebo group (p<0.0001). There was no significant difference between the pain scores during the biopsy itself between the two groups. CONCLUSIONS: Topical diltiazem significantly reduces the pain of probe insertion during prostate biopsy and can be used effectively as an adjuvant to PPNB.

Elevated cytokine responses to Vibrio harveyi infection in the Japanese pufferfish (Takifugu rubripes) treated with Lactobacillus paracasei spp. paracasei (06TCa22) isolated from the Mongolian dairy product.

With the aim of evaluating the effect of a Mongolian dairy product derived Lactobacillus paracasei spp. paracasei (strain 06TCa22) (Lpp) on the cytokine-mediated immune responses to Vibrio harveyi infection, we examined 16 cytokine expressions in the Japanese pufferfish, Takifugu rubripes. Fish were orally treated with the heat-killed Lpp at 1 mg g(-1) body weight d(-1) for 3 days. At 24 h posttreatment, fish were infected by an intramuscular injection of 0.1 mL V. harveyi bacterial suspension (10(8) cfu mL(-1)). Additionally, superoxide anion production (SAP) and phagocytic activity (PA) of head kidney cells were assessed during 120 h postinfection period. Significant up-regulation of pro-inflammatory (IL-1ß, IL-6, IL-17A/F-3, TNF-α and TNF-N), cell-mediated immune inducing (IL-12p35, IL-12p40 and IL-18), antiviral/intra-cellular pathogen killing (I-IFN-1 and IFN-γ), anti-inflammatory (IL-10) and lymphocyte agonistic (IL-2, IL-7, IL-15, IL-21 and TGF-ß1) cytokines was observed in the treated fish compared to control ones during the pathogen infection. Furthermore, significantly increased SAP and PA (P < 0.01; 0.05) were recorded in the treated fish compared to untreated fish. These results suggest the beneficial role of Lpp in enhancement of cytokine-mediated immunity in the Japanese pufferfish against V. harveyi infection and application of this product as a potential fish immunostimulant.

Parathyroid failure following thyroid surgery.

This cross-sectional study was done to see the association of post thyroidectomy parathyroid failure with thyroid disease and type of surgery. It was carried out in the Department of Otolaryngology-Head and Neck Surgery, Bangabandhu Sheikh Mujib Medical University, Dhaka during the period of July 2008 to June 2010. Total 50 cases of thyroid malignancy and multinodular goiter who had undergone total or near total thyroidectomy with or without neck dissection were studied. In this study highest number of cases were found in 3rd decade of age (28%) and there was female predominance (M:F=1:3.54). Overall frequency of post operative hypocalcaemia was 30% (26% was temporary hypocalcaemia and 4% was permanent). Hypocalcaemia revealed clinically in 20% cases and remained subclinical in 10% cases. Hypocalcaemia developed in 42.30% cases of malignant thyroid disease and 16.66% cases of benign thyroid disease (p<0.05). It was found in 54.54% cases with neck dissection and 23.07% cases without neck dissection (p<0.05). Hypocalcaemia developed in 62.5% cases where parathyroid gland were not identified and 23.8% cases where parathyroid gland was identified. Hypocalcaemia developed most commonly on the 2nd post operative day (73.33%). There is a significance difference with development of parathyroid failure after thyroid surgery between benign and malignant thyroid disease and also between thyroid surgery with or without neck dissection.

Cytokine responses in the Japanese pufferfish (Takifugu rubripes) head kidney cells induced with heat-killed probiotics isolated from the Mongolian dairy products.

Cytokine responses in the Japanese pufferfish (Takifugu rubripes) head kidney (HK) cells to heat-killed lactic acid bacteria probiotics isolated from the Mongolian dairy products were investigated by transcriptomic examination. The HK cells were incubated with two heat-killed bacteria, namely Lactobacillus paracasei spp. paracasei (strain 06TCa22) and L. plantarum (strain 06CC2) and the responses of 16 cytokine genes at 0 (control), 1, 4, 8, 12, 24 and 48 h post-stimulation were assayed by multiplex RT-PCR analysis (GenomeLab Genetic Analysis System, GeXPS; Beckman Coulter, Inc.). The 16 genes included in the assay were pro-inflammatory cytokines (IL-1ß, IL-6, IL-17A/F-3, TNF-α and TNF-N), cell-mediated immune regulators (IL-12p35, IL-12p40 and IL-18), antiviral (I-IFN-1 and IFN-γ) and other regulatory (IL-2, IL-7, IL-15, IL-21, IL-10 and TGF-ß1) cytokines. Despite the differences in the transcriptional profiles, expression of all the cytokines tested here was significantly elevated by both the probiotic bacterial stimulants compared with the unstimulated control. Therefore, this in vitro study has demonstrated the modulation of cytokine defense mechanisms in the HK cells by the two heat-killed probiotics indicating their potentiality as novel immunostimulants to fish. However, strain-dependent varied expression of important cytokines (cell-mediated immune regulators, antiviral and anti-inflammatory cytokines) suggests better efficacy of L. paracasei spp. paracasei strain as fish immunostimulant. Further in vivo studies to elucidate the cytokine regulation networks will validate our present observations. A careful evaluation of ant-inflammatory properties may be undertaken using single strain to affirm the immunostimulatory capability. Moreover, application timings and frequency to assess the longevity of immunostimulant effects and to make the application cost-effective need to be evaluated before any practical use in aquaculture.