Function and Characterization of the Argonaute2 Gene in the RNA Interference Pathway in the Diamondback Moth

Arch BiochemMol Biol2019;10(4):035-051DOI: 10.26502/abmb.007Archives of Biochemistry and Molecular BiologyVol. 10 No. 4 -December2019. 36AbstractArgonaute2 (AGO2) is a core catalytic component of the RNA-induces silencing complex (RISC) that binds to small guide RNAs containing small interfering RNA (siRNA) and microRNA (miRNA). The guide RNA leads RISC to the complementary mRNA for gene suppression. We cloned the full length cDNA (2193 bp) of the Ago2gene (PxAgo2) from diamondback moth (DBM, Plutella xylostella). The predicted PxAgo2 protein hadan83 kDa molecular weight withatheoretical isoelectric point of 9.39. The phylogenetic tree showed a high similarity of PxAgo2with Bombyx moriAgo2 (BmAgo2). Western blot and RT-qPCR analyses showed a clear increase in the PxAgo2 mRNA and protein expression levels in the egg, 4thinstar larva, pre-pupa, pupa and adult. The double-stranded RNA-mediated RNAi of PxAgo2in DBM larvae was found 3 h after dsRNA injection,and the knockdown level was increased over time up to 36 h. PxAgo2silencing recovered the expression of PxBurs-?(Bursicon-? inDBM) to the normal expression level, which was suppressed bydsPxBurs-?in a DBM cell line. The overexpression of PxAgo2fundamentally enhanced the PxBurs-? silencing efficiency in DBM cells. Our findings reveal that PxAgo2is involved in the dsRNA-regulated gene silencing mechanism and performs a crucial function in the RNAi process ofDBM

The value of institutional protocols and focused cardiac ultrasound during a case of ultramassive transfusion

A 53-year-old female was admitted to the emergency department with an exsanguinating bleed from the rectum which was of unclear origin. In what could be considered an ultramassive transfusion, 60 units packed red blood cells, 23 units fresh frozen plasma, 20 units platelets, 6 units cryoprecipitate, 30 L of crystalloids, 2 L of colloids, and 4 g of tranexamic acid were transfused over the course of 7 h. An arterio-enteric fistula was diagnosed and treated by an interventional radiologist. The patient recovered rapidly thereafter without any major neurologic, pulmonary, cardiac, or hematologic complications.

Statins everyday versus alternate days: Is there a difference in myalgia rates?

Objective Statins are widely used drugs, known to cause myalgia, leading to high discontinuation rates. The objective of our study was to determine the frequency of myalgia in patients on everyday-dose (EDD) regimen with those on alternate-day dose (ADD) regimen. Methods This cross sectional study was conducted in a tertiary care hospital of Pakistan. A sample size of 400 patients between the age of 40–70 years, taking simvastatin 40 mg for at least 6 months or more were selected. Patients with prior musculoskeletal or neuromuscular complains, and family history of muscular disorders were excluded. Subjects were evaluated for myalgia via a self-administered questionnaire, and those complaining of myalgia were then evaluated for serum vitamin D levels. Data was analyzed through SPSS 16.0 and compared using chi square test. Results The overall prevalence of myalgia was 7% (28/400). Frequency of myalgia in patients taking simvastatin everyday (n = 20, 10%) was significantly higher compared to those taking it every alternate day (n = 8, 4%) (p = 0.02). There was no significant difference between the time of onset, nature, severity, type, or location of myalgia between the 2 groups. The most common cited triggering factor for pain was physical exercise. Of the patients experiencing myalgia, 13 (6.5%) from the EDD group and 6 (3%) from the ADD group had low levels of vitamin D. Conclusions ADD regime was better tolerated by the patients than EDD regime. Alternate day therapy, with or without vitamin D supplementation, may be used by the physicians for troublesome muscular complains.

Iatrogenic atrio-esophageal fistula following a video-assisted thoracoscopic maze procedure: Is esophageal instrumentation justified even when the diagnosis is equivocal?

A 74-year-old female underwent an uneventful bilateral thoracoscopic maze procedure for persistent atrial fibrillation with continuous transesophageal echocardiographic (TEE) guidance. She presented six weeks later with persistent fever and focal neurological signs. Computed tomography of the thorax revealed air in the posterior LA, raising suspicion for an abscess versus an atrioesophageal fistula (AEF). Before undergoing an exploratory median sternotomy, an esophagogastroduodenoscopy (EGD) was performed by the surgeon to check for any esophageal pathology. This however, resulted in sudden hemodynamic compromise that required intensive treatment with vasopressors and inotropes. In this case-report, we review the various intraoperative risk factors associated with the development of AEF during cardiac ablation procedures as well as the potential hazards of esophageal instrumentation with TEE, naso- or oro- gastric devices, and/or an EGD when an AEF is suspected.

Frequency of nasal carriage of Staphylococcus Aureus among health care workers at a Tertiary Care Hospital

Objective: To determine the frequency of Methicillin resistance staphylococcus aureus [MRSA] and identification of drug susceptibility for MRSA isolates among health care workers [HCWs] of a tertiary care hospital of South Punjab Pakistan

Methods: We included 225 HCWs including laboratory staff, doctors, nurses and paramedical staff in this cross-sectional study. The study was conducted in Nishtar medical university/Hospital Multan. The study duration was July-2016 to April-2017. HCWs having no signs of infections and any other systemic disease were included in this study. We used sterile nasal swab sticks for sample collection for determination of S. aureus prevalence. All these samples were processed in the laboratory for MRSA, methicillin sensitive S. aureus (MSSA) and for antimicrobial sensitivity of S. aureus. Chi-square test was used for comparison of frequency of MRSA and MSSA between different HCWs by assuming p-Value

Results: There were 65.3 percent [147] female participants and only 34.7 percent [78] male participants. S. aureus was diagnosed in the nasal flora of 24 percent [54] participants, out of which 9.3 percent [21] were MRSA positive and remaining 14.7 percent [33] were MSSA positive. There was no significant difference in frequency of MRSA and MSSA among different HCWs [p-value 0.79]. Amikacin and vancomycin were 100 percent sensitive for MRSA and MSSA. Clindamycin and ciprofloxacin was 80.9 percent [17] and 71.4 percent [15] sensitive for MRSA and 100 percent and 84.8 percent [28] for MSSA respectively. While oxacillin and Cefoxitin were 100 percent [21] resistant for MRSA and sensitive for MSSA

Conclusion: Prevalence of MRSA and MSSA is high among HCWs in Pakistan. Amikacin, vancomycin and clindamycin have high sensitivity for MRSA and can be used for empirical treatment of MRSA in suspected patients

Albendazole in the treatment of hymenolepiasis in school children

Hymenolepiasis is a helminthic and occasionally fatal disease of human imposing heavy economic losses to human society. Present study was aimed to diagnose the school children for the prevalence and control of Hymenolepiasis. A school based cross-sectional analysis of stool samples collected from 188 children aged 06-15 years was carried out [February to June 2016]. Two stool samples were collected from each student before diagnosing and after treatment. The samples were fixed in 10% formalin and observed under the light microscope using the methods of direct smear in Lugol's solution, normal saline and flotation techniques. On the basis of drugs accessibility all the H. nana infected children were divided in to 2- groups. Children in group A were treated with albendazole [bendazol] 400mg once orally, group B was treated with albendazole [zentel] 200mg orally. Eggs per gram of faeces were counted in each group before and after treatment. Of the 188 children, current study reveals only 6.08% [n=18/296] infection with H.nana and 10.5% [n=16/151] were diagnosed with co infections. The % efficacy of albendazole [Zentel] and albendazole [bendazol] against Hymenolepis nana infection was reported as 83% and 75% respectively. Present study was concluded that albendazole [zentel] is the drug of choice for the treatment of hymenolepiasis in children

Development and validation of a stability-indicating RP-HPLC-FLD method for determination of 5-[[4-chlorophenoxy] methyl]-1, 3, 4- oxadiazole-2-thiol; A novel drug candidate

The present study describes the development and validation of a simple high performance liquid chromatographic method for the determination of a novel drug candidate, 5-[[4-chlorophenoxy] methyl]-1, 3, 4- oxadiazole-2-thiol. The stability-indicating capacity of the method was evaluated by subjecting the compound's solution to hydrolytic, oxidative, photolytic, transition metal- and thermal- stress. The chromatographic separation was achieved over a C18 column [Promosil, 5 microm, 4.60 × 250 mm], maintained at 25°C, using an isocratic mobile phase comprising a mixture of acetonitrile and acidified water of pH 2.67 [1:1, v/v], at a flow rate of 1.00 mL/min and detection using a fluorescent light detector [excitation at 250 nm and emission at 410 nm]. The Beer's law was followed over the concentration range 2.50-50.00 microg/mL. The recovery [98.56-100.19%, SD <5%], intraday accuracy and precision [97.31-100.81%, RSD<5%] and intermediate accuracy and precision [98.10-99.91%, RSD<5%] indicated that the method was reliable, repeatable, reproducible and rugged. The resolution and selectivity factors of the compound's peak from the nearest resolving peak, particularly in case of dry heat and copper metal stress, were found to be greater than 2 and 1, respectively, which indicated specificity and selectivity. The compound was extensively decomposed in alkaline-hydrolytic, oxidative, metal- and dry heat- stress. However, the compound in acidic and neutral conditions was resistant to photolysis. The results of the present study indicate that the developed method is specific, selective, sensitive and suitable, hence, may be used for quality control, stability testing and preformulation studies

Iron deficiency anaemia is a serious maternal health issue during pregnancy

Background: Anaemia is commonly found in developing countries associated with nutritional deficiencies and multiple pregnancies. Iron Deficiency Anaemia in pregnancy is a risk for preterm delivery, low birth weight babies and adverse effects on infant development. The current study investigated frequency of iron deficiency anemia and beta thalassaemia trait during antenatal check up among pregnant women visiting a tertiary care hospital in Islamabad

Objectives: To investigate iron deficiency anemia and beta thalassaemia trait among pregnant females visiting a tertiary care hospital in Islamabad

Study design, settings and duration: The retrospective comparative study was done to investigate iron deficiency anemia among pregnant females visiting a tertiary care hospital in Islamabad from May 2006 to April 2009

Subjects and Methods: The study was conducted at National Institute of Health [Department of Hematology] for Complete Blood Count and differential counts using Automated Hematology Analyzer [Sysmex KX-21]. Serum Ferritin levels were measured by Electro-chemiluminescence technique using Elcys Ferritin Kit [Roche, Germany]

Results: Among 359 subjects, 237 [66%] had iron deficiency anemia. The mean age was 27.32 +/- 4.9 years [range 18-50 years], 18 [5%] had beta thalassaemia trait and 104 [29%] had non-beta thalassaemia Trait. Forteen cases had concomitant iron deficiency anemia and beta thalassemia trait. The findings of this study indicate a relatively higher burden of iron deficiency anemia as compared to beta thalassemia trait

Conclusion: Anemia is a neglected problem which needs priority. Routine iron supplementation should be given to women of child bearing age during pregnancy and post partum. Obstetricians have an important role to aware the women of reproductive age about the iron content in a balanced diet and motivation for iron supplements

Frequency of hepatitis B, hepatitis C and human immunodeficiency viruses in internally displaced persons of south Waziristan, Pakistan

Objectives: To investigate the burden of hepatitis B, hepatitis C and human immunodeficiency virus and to explore route of transmission in internally displaced persons of South Waziristan

Methodology: A cross sectional study was carried out in Gomal Medical Collage, Dera Ismail Khan; Mufti Mehmood Teaching Hospital Bannu and Basic Health Unit Zaferabad, Dera Ismail Khan. Total 300 internally displaced persons [IDPs] of South Waziristan who were undiagnosed and presented with non-specific symptoms were included in the study. The data was analyzed in SPSS version 16. Confidentiality of subjects was ensured

Results: In this study 300 IDPs were screened for HBsAg, anti-HCV and anti HIV antibodies. Out of which male were 190 and female were 110. More males were suffering from hepatitis B [10.3%] than hepatitis C [6.7%] or HIV [0.7%] as compared to females [3.3%], [5%] and [0.3%] respectively. A total of 5[1.7%] cases had history of previous surgery and 17[5.7%] had history of tooth extraction

Conclusion: The frequency of HBV was higher as compared to HCV and HIV infections. The frequency of all the three infections was comparatively more in males as compared to females. Most likely Most likely risk factors for acquistion of infection were unhygienic tooth extractions, unsafe blood transfusions and iatrogenic

Steven Johnson Syndrome – A Triad of Mucosal Lesions, Cutaneous Involvement and History of Recent Drug Exposure

Steven Johnson Syndrome is relative a rare but yet an acute life-threatening condition. A middle aged lady presented to us in an emergency condition with high grade fever (104oF), oral mucosal ulcers, purulent discharge from both eyes, photophobia, generalized skin rashes, flank bruises, dyspnea, severe odynophagia along with dysphagia and blisters on both hands and feet. Because of extensive oral thrush, mouth ulcers and purulent discharge from eyes, the patient was initially suspected to be a case of oral candidiasis and bacterial conjunctivitis. However, due to wide spread muco-cutaneous involvement and after obtaining detailed drug history and getting opinion from dermatology and ophthalmology department, a diagnosis of Steven Johnson Syndrome was established. The treatment was started accordingly and the patient responded dramatically to the treatment, got improved and was subsequently discharged.

Evaluation of different formulations of IGRs against Aedes albopictus and Culex quinquefasciatus (Diptera: Culicidae)

Objective: To test the relative efficacy of pyriproxyfen and methoprene on mortality, deformity, inhibition and emergence to adult stages of Culex quinquefasciatus and Aedes albopictus. Methods: Serial dilutions (0.01-0.05 mg/L) of methoprene, pyriproxyfen 0.5 water dispersible granules (WDG) and pyriproxyfen 1.0 WDG were used to assess mortality and inhibition of 3rd instar larvae of Aedes albopictus and Culex quinquefasciatus. Each concentration and control was replicated four times in completely randomized design. Data on larval mortality, growth inhibition, deformities and adult's emergence was recorded weekly. On the basis of best comparative performance, the efficacy of pyriproxyfen 1.0 WDG at 0.1 g/m

Scar pain, cosmesis and patient satisfaction in laparoscopic and open cholecystectomy

Objective: To compare patient-satisfaction, scar-pain and cosmesis between laparoscopic and open-cholecystectomy. Study Design: Cross-sectional survey. Place and Duration of Study: Khyber Teaching Hospital, Peshawar, from August 2012 to May 2014

Methodology: A total of 400 patients, who had undergone open or laparoscopic cholecystectomy in all units of the Surgical Department, were included. Data was collected on questionnaires given and read to the patients along with counselling and information regarding scar-pain using visual analog score, and satisfaction and cosmesis on a 0 - 10 scale, by a medical professional in the patients' native language. This was done postoperatively on patients' follow-up visits at 1 and 4 weeks

Results: Mean scar pain score at 1 and 4 weeks postoperatively was higher for open-cholecystectomy; 4.96 +/- 1 and 0.96 +/- 1, compared to 2.24 +/- 0.6 and 0, respectively for laparoscopic-cholecystectomy [p < 0.001 and < 0.001]. Cosmesis was higher for laparoscopic-group; 8.6 +/- 1.2 vs. 6.2 +/- 1.46 for open-cholecystectomy [p < 0.001]. Patient-satisfaction was higher for laparoscopic-cholecystectomy; 9.28 +/- 1.5 vs. 8.32 +/- 2.3 for open-cholecystectomy [p < 0.001]. Mean-cosmesis score was higher for laparoscopic-cholecystectomy for those younger than 40, females and unmarried. Mean patientsatisfaction score was higher for those older than 40 years who had undergone open-cholecystectomy, women who had undergone laparoscopic-cholecystectomy and for unmarried patients who had laparoscopic-cholecystectomy

Conclusion: Overall patient-satisfaction and cosmesis scoring was higher for laparoscopic-cholecystectomy especially among females, unmarried and younger than 40 years. Patients of 40 years and older had greater satisfaction scoring for open-cholecystectomy. Therefore, laparoscopic-cholecystectomy should be favoured in females and unmarried patients and those younger than 40 years

Prophylactic rectal nsaids in the prevention of post-ercp pancreatitis

Objective: To evaluate the effectiveness of rectal non-steroidal anti-inflammatory drugs [NSAIDs] using diclofenac in preventing pancreatitis following ERCP

Methodology: It was a randomized, double blinded, placebo controlled study carried out at Surgical Unit-l, Holy Family Hospital, Rawalpindi from May 2013 to April 2014. A total of 108 patients were included and randomly assigned in each of study and placebo groups. Group I patients received 100 mg of diclofenac per rectally while group II patients received a glycerine suppository per rectally [placebo] before the start of ERCP. Post ERCP pancreatitis [PEP] was diagnosed by clinical evaluation and raised serum amylase levels after four hours of the procedure in both groups [study vs control]

Results: Out of 108 patients 32 were males and 76 were females. Mean age was 46.09 +/- 12.31 in group I, while it was 42.93 +/- 14.69 in Group II. Mean serum amylase level, 4 hours after ERCP, was 184.70 +/- 36.34 in the diclofenac group, while it was 388.20 +/- 57.27 IU/L in the control group. Thirty one patients were diagnosed with Post ERCP pancreatitis, out of which nine patients belonged to the study group and twenty two were of the control group [P = 0.000]

Conclusion: Per-rectal administration of diclofenac suppository prior to ERCP results in significant reduction in the frequency of ERCP induced pancreatitis

Fundamentals of multitier medical support planning for low intensity conflict in Pakistan

This study was aimed to visualize the frequency and type of imminent terrorist events and plan a flexible medical support hierarchy capable of effectively responding to terrorist scenarios with focused, adaptive and efficient medical support doctrines. Efficient medical support system reduces mortality, raises morale and neutralizes the effects of terrorist incidents. Medical support in low intensity conflict [LIC] is necessary to be revitalized in line with modern operations requirements. After thorough literature search all available standing operating procedures [SOP], were reviewed for medical support in LIC. To be effective, the medical support system ought to be very well carved, well rehearsed, ever evolving, innovative and practical. It is ought to be flexible and futuristic to accommodate difficult challenges, yet plausible and practical enough to conform to the ground realities. Deliberate and focused endeavors at multiple tiers involving diverse organizations with singularity of aim and purpose i.e. to render sickness preventive, health promotive and disease curative facilities where ever the nation calls in line with the splendid traditions of Pakistan Armed Forces

Receiver operator characteristic curve analysis of the Lintula score for reduction of negative appendectomies in adults

To evaluate the Lintula score in reducing negative appendectomies in the adult population. Descriptive analytical study. Surgical Department, Khyber Teaching Hospital, Peshawar, Pakistan, from August 2012 to April 2014. A total of 408 emergency patients with a clinical diagnosis of acute appendicitis were included in the study. True or negative appendectomy status was determined per-operatively. Lintula score was calculated afterwards and evaluated for various cut-off points. Among the study population, 72 [17.6%] had a normal appendix by operative assessment and 336 [82.4%] had an acutely inflammed appendix. The receiver operating characteristic curve showed that the optimal cut-off point was

Laparosopic assisted Right Hemicolectomy; an initial experience

Experience with Laparosopic assisted Right Hemicolectomy is presented. Prospective study. Surgical Unit-I, Holy Family Hospital, Rawalpindi, Pakistan. 2010 to 2014. 20 patients underwent laparosopic assisted right hemicolectomy. Duration of operation, postoperative pain, duration of post-operative analgesia, and frequency of surgical site infection and length of hospital stay were noted. There were 13 male and 7 female patients with the age range of 25-70 years [45 +/- 11years]. Eleven [11] patients were suffering from carcinoma colon wheras nine [09] were suffering from Tuberculosis. Mean operative time was about 92 minutes. Only four opiod injections were required as post-operative analgesia. Oral intake was started after 24 hours. The mean length of hospital stay was 5.5 days with no case of surgical site infection. Laparoscopic assisted right hemicolectomy is a safe and viable option in our setup

Urothelial carcinoma of urinary bladder; immunohistochemical demonstration of COX-2 in correlation with various grades

In spite of the fact that inflammation has been regarded as a localized or generalized defensive component of the body to different types harmful stimuli, there has been becoming confirmation of its strong part in initiation or progression of different ailments particularly related with cancer

Objectives: Aim of this study was to recognize the pattern of expression and level of intensity of COX-2 in different grades of papillary urothelial carcinoma of urinary bladder along with significance of COX 2 in tumerogenesis of urothelial carcinoma of urinary bladder

Period: 1.1.2009 to 31.12.2012

Methods: The marker of COX-2 was investigated by using Immuno- histochemistry

Results: COX 2 was not detected in normal urothelium, but its intensity was expressed as 68% in low grade, 72% in high grade and 80% in invasive urothelial carcinoma

Conclusion: Results of the present study indicate that COX-2 as a component of inflammation play an important role in progression of urinary bladder tumor and encourage use of COX 2 inhibitors as potential antitumor agent

If it's itchy, its scabies not always

Scabies is commonly characterized by the presence of itchy rash but this typical symptom does not necessarily always mean scabies. A female infant presented to the paediatrics department of Kuwait teaching hospital with a characteristic pruritic rash on soles suggestive of scabies. However, failure of the anti-scabies treatment, absence of positive personal and hygienic history and progression towards the deterioration of existing situation lead to the establishment of a history and clinical based diagnosis of Infantile Acropustulosis. The treatment was started accordingly and soon after starting treatment, the patient showed the signs of recovery. No doubt that scabies is comparatively more common as compared to infantile acropustulosis but yet scabies must not be considered as an absolute diagnosis for a patient presenting with an itchy rash particularly on soles

study of the spectrum of presenting symptoms, diagnostic investigations, treatment and outcome of severe malaria in the admitted children of Kuwait Teaching Hospital ,Peshawar, Pakistan

Objectives: to document the spectrum of presenting symptoms, diagnostic investigations, treatment and outcome of severe malaria in the hospitalized children of Kuwait teaching hospital

Methodology: this was a descriptive study using patient case records of children admitted from 1[st] January till 31[st] December 2013. All children with malaria requiring admission and treatment with parenteral anti-malarial were categorized as severe malaria and included in the study. Data was analyzed using Microsoft office Excel 2007

Results: 32 cases fulfilled the inclusion criteria. Fever was the commenest presenting symptoms present in 100% of cases [n=32]. Fifty percent children [n=16] had pallor, 12.5%[n= 4] cough, 9.3%[n=3] had dysuria and 6.25%[n=2] had fits recorded as additional presenting symptoms. 71.8% [n=23] had parasitological confirmation test before commencement of treatment. 100% of this parasitological confirmation was done by peripheral blood smear/microscopy test. Out of the 23 who had microscopy done 60.86%[n=14] had negative result and 39.13%[n=9]were positive for plasmodium of which,88.88%[n=8]were vivax and 11.11%[n=1] was falciparum.93.75%[n=30] of the parenteral anti-malarial used was intravenous quinine. All except one child 96.8%[n=31] recovered with the anti-malarial treatment used

Conclusions: fever and pallor are the two most common presenting symptoms of hospitalized children with severe malaria. Microscopy remained the most common investigation for the diagnosis of malaria. Quinine is the most frequently used drug for severe malaria. The study highlighted the need to further improve the case management of severe malaria in children