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CRISPR-Cas9 in agriculture: Approaches, applications, future perspectives, and associated challenges

Abstract

The discovery of an adaptive immune system especially in archae and bacteria, CRISPR/Cas has revolutionized the field of agriculture and served as a potential gene editing tool, producing great excitement to the molecular scientists for the improved genetic manipulations. CRISPR/Cas9 is a RNA guided endonuclease which is popular among its predecessors ZFN and TALEN’s. The utilities of CRISPR from its predecessors is the use of short RNA fragments to locate target and breaking the double strands which avoids the need of protein engineering, thus allowing time efficiency measure for gene editing. It is a simple, flexible and highly efficient programmable DNA cleavage system that can be modified for widespread applications like knocking out the genes, controlling transcription, modifying epigenomes, controlling genome-wide screens, modifying genes for disease and stress tolerance and imaging chromosomes. However, gene cargo delivery system, off target cutting and issues on the safety of living organisms imposes major challenge to this system. Several attempts have been done to rectify these challenges; using sgRNA design software, cas9 nickases and other mutants. Thus, further addressing these challenges may open the avenue for CRISPR/cas9 for addressing the agriculture related problems.

Open access
Production and Marketing of Carp Species in Rupandehi, Nepal

Abstract

The research was conducted from January 2019 to June 2019 to study the fish production and marketing system in fish zone, Rupandehi. Altogether 95 respondents including 27 small farmers, 35 medium farmers and 33 large farmers were selected for household survey using simple random sampling for the study. Statistical tools, Ms- excel and SPSS of version 23 were used for data analysis. From the study, it was evident that, six species of carps of fingerlings size were stocked with high average number of common carps (3885.5/ha.) followed by silver carp (3880.5/ha). The average production of fish species was 6.06ton/ha which is slightly higher than the national average. The observed data revealed that, marketing of major quantity of fish was from on farm sale i.e. 55.78% followed by local market sale (18.94%), live fish sale in retailers (5.26%). Similarly, percentage of farmers selling fish to wholesaler, consumers, retailers and neighbours were 63.3%, 30.5%, 26.5% and 26.3% respectively. Cost and returns analysis showed average B/C ratio of 2.13. Problem ranking using 5 point scale value showed high cost of feed as the most serious problem with index value of 0.94 in the study area.

Open access
Professional education among haemophilia nurses: a survey of current practices

Abstract

Background: Guidance from the European Association for Haemophilia and Allied Disorders (EAHAD) sets out the educational milestones haemophilia nurses should aim to achieve. However, little is known about the resources nurses use for education and current awareness.

Aims: To assess the current educational level of haemophilia nurses, how and where they access ongoing education, where they feel they need extra support, and how best this teaching could be delivered.

Methods: Haemophilia nurses in the Haemnet Horizons group devised and piloted a questionnaire. This was distributed in hard copy to nurses attending the 2019 EAHAD Congress and promoted as an online survey hosted by Survey Monkey.

Results: Seventy-five replies were received from nurses in Europe (46 in the UK), and two from nurses in Chile and the Philippines. Most described their role as ‘specialist nurse’, with the majority having worked in haemophilia care for up to ten years. Half had a nursing degree and one quarter had a nursing diploma. Three quarters had attended at least one course specifically related to haemophilia nursing. Almost all used academic sources, study days and the websites of health profession organisations as information sources. Most also used Google or Wikipedia, but fewer used Twitter. Patient association websites were more popular among non-UK nurses. About half attended sponsored professional meetings and three quarters reported that educational meetings were available in their workplace. A clear majority preferred interactive and face-to-face activities using patient-focused content.

Conclusions: The study shows that nurses, predominantly in Western Europe, access a range of educational resources, most of which are ‘traditional’. Use of online sources is high, but social media are less popular than Google or Wikipedia. Further research is needed to explore the potential of new media for haemophilia nurse education, and whether the current educational levels and needs highlighted in the survey remains the same across the whole of Europe.

Open access
Assessment of subclinical hypothyroidism for a clinical score and thyroid peroxidase antibody: a comparison with euthyroidism grouped by different thyroid-stimulating hormone levels

Abstract

Background

Subclinical hypothyroidism (SCH) might have many symptoms of hypothyroidism. The controversy appears to lower the level of thyroid-stimulating hormone (TSH) and group subjects with TSH of more than 3 or even 2.5 mIU/L as SCH subjects.

Objectives

To assess SCH subjects both clinically using Zulewski clinical score and biochemically and to evaluate whether the euthyroid subjects with high-normal TSH (HNT) have any clinical symptom or subnormal biochemical finding.

Methods

A prospective cross-sectional study of 233 subjects, 67 with SCH and 166 euthyroidism, was conducted. Euthyroid subjects were divided according to the level of TSH as HNT (>2.5 mIU/L) and low-normal TSH (0.5–2.5 mIU/L). The subjects were examined for clinical feature including Zulewski clinical score and biochemical evaluations including thyroid peroxidase antibody (TPO-Ab) titer. The comparisons between groups were assessed using independent sample t test, and correlations between variables were evaluated using Pearson correlation.

Results

A significantly higher clinical score and higher frequencies of symptoms were found in the SCH group compared to the euthyroid group. The most frequent symptom was fatigue. Euthyroid subjects with HNT were found to have higher TPO-Ab titers than those with low-normal TSH, P < 0.05. The Zulewski clinical score was positively correlated with TSH and TPO-Ab titer but negatively correlated with the FT4 level, P < 0.05.

Conclusions

Zulewski clinical score is higher in SCH subjects compared to euthyroid subjects and can aid in assessing SCH subjects. A significant correlation exists between Zulewski clinical score and each of the TSH, FT4, and TPO-Ab titer levels. The frequency of TPO-Ab positivity is high in SCH. Additionally, euthyroid with higher TSH levels has higher level of TPO-Ab titer but not higher clinical score.

Open access
Clinical outcomes and surgical preferences for breast-conserving surgery and mastectomy: a propensity score-matched analysis

Abstract

Background

While numerous randomized controlled trials have demonstrated long-term survival rates for patients with early-stage breast cancer treated with breast-conserving surgery (BCS) comparable to mastectomy, the latter remains the most prevalent surgical option to treat early-stage breast cancer in Thailand.

Objectives

To investigate the potential determinants affecting the decision on selecting BCS or mastectomy for the treatment of early-stage breast cancer and to compare the disease-free survival and overall survival between the treatments using a propensity score-matched analysis.

Methods

Patients diagnosed nonmetastatic breast cancer at the Queen Sirikit Breast Cancer Center from January 2006 to December 2015, were retrospectively identified and grouped intro patients who received BCS or mastectomy. After propensity score matching, 356 BCS and 209 mastectomy patients were identified, and statistical analysis was conducted to determine treatment selection factors and compare disease-free and overall survival.

Results

Disease-free survival and overall survival in months comparing BCS and mastectomy were not statistically different with P values of 0.11 and 0.77, respectively. Determinants of treatment selection found that younger age, surgeon preference, smaller tumor size, and lower tumor grade were statistically significant factors in the selection of BCS over mastectomy. The majority of surgeons had a preference for one treatment over the other (P < 0.001).

Conclusion

The outcome of BCS is comparable to mastectomy in early-stage breast cancer patients. Key determinants affecting the selection of treatment were identified to be patient age, characteristics of the tumor, and surgeon’s preference.

Open access
Common orthopedic problems in the neonate: a comparative study of 2 periods at a tertiary-care hospital

Abstract

Background

In 1975, King Chulalongkorn Memorial Hospital (KCMH) in Bangkok, Thailand, published data on common orthopedic problems in neonates.

Objectives

To determine the more recent incidence of these conditions and to compare the results with those reported 40 years ago by KCMH. The data were also compared with a recent report from Siriraj Hospital.

Methods

We reviewed medical records of newborn infants from 2012 to 2016, all of whom were born in KCMH with deformities of upper and lower extremities or other birth-related injuries. The cases were grouped according to International Classification of Diseases, 10th edition. The incidence of common neonatal orthopedic problems was calculated as cases per 1,000 live births and compared, using a chi-squared test, with the earlier data from KCMH and with the data recently reported by Siriraj hospital.

Results

Of the 24,825 live births, 54% were male and 46% were female. The average birth weight was 3,052.2 ± 516.1 g (range, 535–5,320 g) for infants. The most common deformity was postural clubfoot, followed by calcaneovalgus and hip dysplasia, with incidences of 1.37, 0.93 and 0.52 per 1,000 live births, respectively. The incidence of calcaneovalgus, metatarsus adductus, and clubfoot was lower than in the 1975 study and in the report from Siriraj. In all 3 studies, the incidence of birth-related injury was similar and clavicular fracture was the most common, with a rate of 1.3/1,000 live births.

Conclusion

The incidence of common neonatal orthopedic problems varies among institutions. Birth-related injury is a major challenge. The results may be utilized as updated data and as a starting point for parental education.

Open access
Conservative surgical management for immediate postpartum hemorrhage

Abstract

Background

Conservative surgical management for postpartum hemorrhage (PPH), such as balloon tamponade, uterine compression suture, and uterine artery ligation, has the benefit of preserving reproductive function.

Objectives

To assess the efficacy and subsequent pregnancy outcome of conservative surgical management for patients with immediate PPH.

Methods

Medical records of patients who had PPH between January 2011 and December 2016 were reviewed. Conservative surgical management included B-Lynch uterine compression suture, Bakri balloon tamponade, and uterine artery ligation. The treatments were considered successful if patients did not require subsequent hysterectomy. Perioperative complications and subsequent pregnancy outcomes were recorded.

Results

Of 30,271 deliveries, 669 patients experienced PPH or 2.2% of total deliveries. Sixty-one patients (9.1%) did not respond to medical treatment with various uterotonic agents. Hysterectomy was selected initially in 30 patients. Conservative surgical management was performed in 31 patients: 15 Bakri balloon tamponade, 13 uterine compression suture, and 3 uterine artery ligation. There were 3 patients who failed Bakri balloon tamponade and proceeded to perform uterine compression suture with successful outcome. The success rates for conservative surgical treatment were 66.7%, 75%, and 66.7%, respectively. All patients who had successful conservative surgical management resumed normal menstruation. Three out of 11 patients (27.3%) who desired subsequent pregnancy were able to conceive and carry out a viable pregnancy.

Conclusion

Conservative surgical management has acceptable success rates for controlling intractable immediate PPH. Implementation of such procedures should be done to preserve fertility and decrease maternal morbidity and mortality.

Open access
Distinguishing magnetic resonance imaging features between idiopathic hypertrophic pachymeningitis and secondary hypertrophic pachymeningitis

Abstract

Background

Hypertrophic pachymeningitis (HP) is a rare chronic inflammatory disorder characterized by marked fibrous thickening of the cerebral and/or spinal dura mater. This condition is caused by infection, inflammation, autoimmune disorder, neoplasms, or idiopathic. Magnetic resonance imaging (MRI) may play an important role in differentiating idiopathic HP from secondary HP, may avoid unnecessarily invasive dural biopsy, and prompt specific treatment.

Objective

To determine the specific MRI findings for differentiation between idiopathic HP and secondary HP.

Method

A total of 34 patients underwent MRI of the brain and cervical spine from January 2003 to December 2015. In all, 23 patients were diagnosed idiopathic HP and 11 patients were secondary HP. Demographic data and imaging findings reveal the following: configuration, thickness, signal intensity on T1-weighted image (T1WI), T2-weighted image (T2WI), and enhancement pattern of the lesions. The data were analyzed by T-test and Fisher’s exact test.

Result

Secondary HP were significantly located at anterior and middle cranial fossa (P = 0.033). There is no significant difference of lesions in configurations, T1 and T2 signal intensity and patterns of enhancement. There was significant and exclusive difference in T2 hypointense/dark intensity and homogeneous enhancement in idiopathic HP (75%, P = 0.044).

Conclusions

MRI may play a complimentarily important role in distinguishing idiopathic HP from secondary HP. Idiopathic HP is probably preferred diagnosis in the lesions with T2-rim pattern and T2 hypointense/dark intensity with homogeneous enhancement.

Open access
Changes in protein patterns of Staphylococcus aureus and Escherichia coli by silver nanoparticles capped with poly (4-styrenesulfonic acid-co-maleic acid) polymer

Abstract

Background

While silver nanoparticles (AgNPs) are increasingly attractive as an antibacterial agent in many applications, the effect of AgNPs on bacterial protein profiles, especially AgNPs stabilized by polymeric molecules, is not well understood.

Objectives

To investigate the changes in bacterial protein patterns by AgNPs capped with poly (4-styrenesulfonic acid-co-maleic acid) (AgNPs-PSSMA) polymer toward Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922.

Methods

The growth of bacteria after incubated with AgNPs-PSSMA for different time intervals was determined by optical density at 600 nm. Their protein patterns were observed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and the proteomic analysis of extracted proteins was determined by liquid chromatography-tandem mass spectrometry (LC–MS/MS).

Results

AgNPs-PSSMA was able to inhibit the growth of both S. aureus and E. coli cells. The treated bacterial cells expressed more proteins than the untreated cells as seen from SDS-PAGE study. Nanosilver (NS) caused the upregulation of metabolic gene, waaA, in S. aureus cells. For E. coli cells, the upregulated proteins were metabolic genes (srlB, fliE, murD) and other genes dealt with DNA replication (dinG), DNA–RNA transcription (yrdD), RNA– protein translation (rplD), molecular transport (sapF), and signal transduction (tdcF).

Conclusions

The antibacterial effect of AgNPs-PSSMA may arise by changing the bacterial proteins and thus interfering with the normal cell function.

Open access