Effect of quercetin on the number of blastomeres, zona pellucida thickness, and hatching rate of mouse embryos exposed to actinomycin D: an experimental study

Background: Quercetin is a flavonoid with the ability to improve the growth of embryos in vitro, and actinomycin D is an inducer of apoptosis in embryonic cells

Objective: The aim was to evaluate the effect of quercetin on the number of viable and apoptotic cells, the zona pellucida [ZP] thickness and the hatching rate of preimplantation embryos exposed to actinomycin D in mice

Materials and Methods: Two-cell embryos were randomly divided into four groups [Control, Quercetin, actinomycin D, and Quercetin + actinomycin D group]. Blastocysts percentage, hatched blastocysts, and ZP thickness of blastocysts was measured. The number of blastomeres was counted by Hoechst and propidium iodide staining and the apoptotic cells number was counted by TUNEL assay

Results: The results showed that the use of quercetin significantly improved the growth of embryos compared to the control group [p=0.037]. Moreover, quercetin reduced the destructive effects of actinomycin D on the growth of embryos significantly [p=0.026]

Conclusion: quercetin may protect the embryos against actinomycin D so that increases the number of viable cells and decreases the number of apoptotic cells, which can help the expansion of the blastocysts, thinning of the ZP thickness and increasing the hatching rate in mouse embryos

Multiplex real-time RT-PCR assay for transfusion transmitted viruses in sero-negative allogeneic blood donors: an experience from Southern Pakistan

Background: Blood transfusion safety commences with healthy donor recruitment. The threat of transfusion transmitted infections is greatly minimized by serological tools but not entirely eliminated. Recently, nucleic-acid testing for blood donor screening has virtually eliminated this jeopardy. Methods: This prospective study was conducted from February 2015 to February 2016. Samples from seronegative donors were run on multiplex assay (Cobas, S-201 system platform, Roche) in a batch of six [MP-NAT]. In case of reactive pool, tests were run on every individual sample [IDNAT]. Results: Of 16957 donors, 16836 (99.2%) were replacement donors and the remaining 121 (0.7%) were voluntary donors, with a mean age of 29.09 ± 7.04 years. After serologic screening of all 16957 donors, 955 (5.6%) were found to be reactive; 291(1.71%) were reactive for hepatitis-B surface antigen, 361 (2.12%) for antibody to hepatitis C virus (anti-HCV), 14 (0.08%) for antibody to human immunodeficiency virus, 287 (1.69%) for syphilis and 2 (0.01%) for malaria. 14 (0.08%) NAT reactive donors were identified after testing the 16002 seronegative donors, with an overall NAT yield of one reactivity out of 1143 blood donations; 10 donors for HBV-DNA (HBV NAT yield-1:1600) and remaining 4 for HCV-RNA (HCV-NAT yield-1:4000). None were HIV positive. Conclusion: NAT has improved the safety attributes in blood products. Although the positivity rate for NAT testing is low but in view of the high prevalence of transfusion transmitted infections in our country, we recommend the parallel use of both serology and NAT screening of all donated blood.

Adverse reactions in allogeneic blood donors: a tertiary care experience from a developing country

Objectives: Fragmented blood transfusion services along with an unmotivated blood donation culture often leads to blood shortage. Donor retention is crucial to meet the increasing blood demand, and adverse donor reactions have a negative impact on donor return. The aim of this study was to estimate adverse donor reactions and identify any demographic association

Methods: We conducted a prospective study between January 2011 and December 2013. A total of 41,759 healthy donors were enrolled. Professionally trained donor attendants drew blood and all donors were observed during and following donation for possible adverse events for 20 minutes. Blood donors were asked to report if they suffered from any delayed adverse consequences

Results: Out of 41,759 blood donors, 537 [1.3%] experienced adverse reactions. The incidence was one in every 78 donations. The mean age of donors who experienced adverse events was 26.0 +/- 6.8 years, and all were male. Out of 537 donors, 429 [80%] developed vasovagal reaction [VVR], 133 [25%] had nausea, 63 [12%] fainted, 35 [6%] developed hyperventilation, 9 [2%] had delayed syncope, and 9 [2%] developed hematoma. Arterial prick, nerve injury, cardiac arrest, and seizures were not observed. Donors aged less than < 30 years and weighing <70 kg were significantly associated with VVR, hyperventilation, and nausea [p < 0.005]. Undergraduates and Urdu speaking donors also had a significant association with fainting and nausea, respectively [p < 0.05]

Conclusion: The prevalence of adverse events was low at our tertiary center. A VVR was the predominant adverse reaction and was associated with age and weight. Our study highlights the importance of these parameters in the donation process. A well-trained and experienced phlebotomist and pre-evaluation counseling of blood donors could further minimize the adverse reactions

Thyroidectomy under local anesthesia

Objective: to find effectiveness of thyroid surgeries under local anaesthesia when the patients had contraindication to general anesthesia

Study design: descriptive case series

Place and Duration of study: department of Surgery, Jinnah Hospital / Allama Iqbal Medical College Lahore, from January 2004 to March 2015

Methodology: a total of 100 patients were included. For infiltration anesthesia, lignocaine and bupivacaine were used. Two ampoules of lignocaine and two ampoules of bupivacaine [40 ml each] were taken and 160 ml normal saline was added in it. Total amount of infiltrate was 200 ml into which 0.5 ml of adrenaline 1/1000 was added. Total adrenaline concentration came to be 1/400000 in 200 ml. Before draping the patient, local anaesthesia was infiltrated starting from the incision, to all the area below from suprasternal notch to above the thyroid cartilage and laterally to anterior border of sternocleidomastoid. Patients were given tablet alprazolam [Xanax] 0.5 mg six hours prior to surgery. Just before the operation intramuscular injection diclofenac [Dicloran] 75 mg and intravenous antibiotics were also administered

Results: the study population included 81 females and 19 males with the mean age of 35+/-5 year. The mean procedure time was 105 minutes. Procedures included subtotal bilateral thyroidectomy [n=31], lobectomy [n=46] and total thyroidectomy [n=23]. Postoperative complications included bleeding [n=2]. Patients showed full self-dependency within 4 to 6 hours and remained under medical surveillance for 48 hours

Conclution: use of local anesthesia in thyroid surgery makes it cost effective with less postoperative pain. Early smooth recovery was also observed. Thyroid surgeries under local anesthesia may be considered a safe alternative where general anesthesia is not available or contraindicated for medical reasons

Frequency of HBV, HCV andmalaria infections in cytopenic patients coming for bone marrow aspiration to bacha khan medical college mardan

Objective: To know the frequency of HCV, HBV, and Malaria infections in cytopenic patients coming for Bone Marrow Aspiration

Study Duration: Observational study

Place and Duration of Study: This study was conducted in Pathology and Medicine Departments of Bacha Khan Medical College Mardan from January 2012 to December 2014

Materials and Methods: 100 patients with cytopenia were referred for Bone Marrow Aspiration and were screened for HBV, HCV and malaria infections. Complete blood counts were performed on 50 healthy individual as a control group.Both Thick and Thin Blood smears were examined for Malaria, while Anti HCV and HBs Ag Screening was done by kit method [Immunochromatographic Method SD company]. Complete blood count was performed by Heamatologyanalyser. [Sysmex Hematology Analyzer ]

Results: 4 out of 100 cytopenic patients were Anti HCV positive, who were referred for Bone Marrow Aspiration. These patients had a Hemoglobin level 10.5 +/- 0.978 G/dl, Platelets counts 120 +/- 24.768×103/uL. 2 out of 100 Cytopenic patients were HBs Ag Positive. They had anemia and thrombocytopenia, Hb level was 10.4 +/- 0.879 g/dland platelets count was 119 +/- 24.965×103/uL respectively. while 7 [7%] out of 100 cytopenic patients had malaria microscopy.TheypresentedwithHbof9.5 +/- 0.978g/dl,plateletcount120 +/- 24.768×103/uLandTLC3.8 +/- 1.365×103/uL respectively. Counts in HCV, HBV and malaria are significantly lower than the control groups value < 0.0024

Conclusion: Cytopenia is a significant finding in HCV, HBV and Malaria infections and patients presenting with any cytopenia patients referred for Bone marrow Aspiration presenting with cytopenia should be properly screened for HCV, HBV, and Malaria as these can be the cause of cytopenia thus unnecessary use of Bone Marrow Aspiration can be presented

Caudal epidural steroids; outcome in lumber disc herniation

Introduction: About 85% of the population experience low back pain during adulthood and annual incidence is 5 to 15% with no gender discrimination

Design: Quasi intervention experimental study

Setting: DHQ teaching Hospital and at Fazil Memorial Hospital Gujranwala

Period: November 2010 to November 2012 [two years]

Material and Methods: Hundred patient fulfilling the inclusion criteria with SYMPTOMATIC LUMBER DISC herniation were selected. Efficacy was determined by improvement in Denis Pain scale and Mcnab's functional criteria. Safety was determined by absence / rareness of complications i.e spinal anesthesia, transient hypotension, and paresthesia, difficulty in voiding, infection and meningitis. Follow up was weekly for 6 weeks

Results: Out of hundred cases 53 were gents and 47 were ladies. Mean age was 37.56 [22-50 yrs]. Maximum cases 72% were in third decade of life. Maximum pain relief was noted after three weeks of first injection, 72% of the cases were either having no or mild pain. Functional recovery was excellent in 44% and good in 40% of the patients. Complications were fewer and mild, Spinal anesthesia in 2%, transient hypotension in 3%. Paresthesia in 8% and 10% of the patient developed supra pubic fullness but only one needed catheterization. Infection and meningitis was not observed in any case

Conclusions: Caudal epidural steroids injection is safe and effective mode of treating lumbar disc herniation

Pattern of skin malignancies in patients presenting to dermatology department Hayatabad Medical Complex Peshawar

This study was conducted to determine the pattern of cutaneous malignancies among patients attending the skin out-patient department Hayatabad Medical Complex Peshawar. This was descriptive analysis of skin cancers carried from January 2011 to December 2011. Histologically diagnosed skin cancers were reviewed and analyzed according to age, gender, site of distribution and histological types. Hundred histologically diagnosed skin malignancies were reported during the period under review. Sixty cases [60%] were males and 40 cases [40%] females with a male to female ratio of 1.5:1. Age ranged from 22 years to 86 years. Mean age was 57.05 +/- 11.39 SD. Mean duration of disease in months were 16.78 +/- 9.07SD. Basal cell carcinoma [BCC] was the most common malignancy consisting of 32[32%] cases followed by Squamous cell carcinoma [SCC 18%] with 10 [10%] cases of melanoma and 9[9%] cases of Mycosis Fungoides. Actinic Keratosis, Kaposi's sarcoma, Bowen's disease, Dermatofibroma, Atypical fibroxanthoma constituted the remaining. The most common skin malignancy was BCC followed by SCC and melanoma. Larger studies should be conducted to ascertain the actual prevalence of skin tumors

Skull base osteitis: our experience and systematic review of literature

Objective: this article discusses etiopathogenesis, diagnostic problems and various management modalities available to manage skull base osteitis [SBO] which is a rare but life threatening disorder and in addition national and international references will be reviewed

Material and Methods: cross sectional, retrospective study carried out at ENT unit-SIMS/SHL which included 17 patients with SBO which were sorted out for etiological diagnosis based on detailed history, physical examination and labs

Results: mean age was 52 years. Diabetes is the most significant risk factor and was seen in 82% of patients, pseudomonas was isolated in 8 patients. The main complication facial paralysis was present in 64% of patients

Conclusion: SBO remains a severe debilitating and life-threatening condition. It may develop in patients with benign otitis media and externa, and must be considered in all patients with temporal bone inflammation; especially those with risk factors and those who fail to improve with more conservative measures. Small-vessel vasculopathy and immune dysfunction associated with diabetes are primarily responsible for this predisposition. Cranial nerves most commonly the facial, can be affected by inflammation along the skull base or by a neurotoxin produced by Pseudomonas species. We, in this article, intend to share our experience in managing seventeen patients with SBO over a period of three years and review the relevant and recent global literature suggesting updates

Short term effects of rosuvastatin on plasma concentration of high sensitivity c-reactive protein in patients with chronic stable angina

To determine the short term effects of rosuvastatin on elevated base line high-sensitivity C-reactive protein [hs-CRP] in patients with chronic stable angina. This Quasi-experimental comparative study was conducted in Cardiology department, Lady Reading Hospital Peshawar, between March 2010 and February, 2011. We selected 44 consecutive patients age 40 years or above, of any gender having hs-CRP levels >/= 1.2mg/l with chronic stable angina. Base line levels of hs-CRP, total cholesterol, LDL cholesterol, HDL cholesterol, triglycerides, and creatine phosphokinase [CPK] were measured in fasting status. These patients were treated with rosuvastatin 20 mg once daily at night and followed up for one month. Using SPSS version 16 data was analyzed. Mean age was 53 +/- 7.2 and 50% were females. Following treatment with rosuvastatin 20mg for one month the mean hs-CRP levels reduced from 4.08 +/- 2.56 to 2.72 +/- 2.40 [95%CI, 0.41 to 2.29, p=0.006]. Similarly mean total cholesterol levels decreased from 185.88 +/- 37.62 to 147.45 +/- 38.35, [p=0.0001]. LDL cholesterol decreased from 118.34 +/- 31.31 to 86.63 +/- 35.72 [p= 0.0001]. But mean HDL cholesterol had no significant increase from baseline levels i.e. from 32.18 +/- 9.93 to 33.95 +/- 7.65 [p=0.174]. TGs levels reduced from 240.11 +/- 123.66 to 197.43 +/- 88.24 [p=0.008]. Mean CPK levels did not differ significantly from base line at follow up, from 101.43 +/- 58.63 to 96.22 +/- 55.10 [p=0.646]. Short term treatment with rosuvastatin significantly decreases elevated hs-CRP levels in patients with chronic stable angina

Safety in functional endoscopic sinus surgery [FESS]

To document the peroperative and postoperative complications of functional endoscopic sinus surgery. 113 patients diagnosed as having sinonasal polyposis who underwent FESS at ENT unit 1 SIMS/SHLfrom September 2010 to June 2012, were registered in this study. Out of 113 patients 15 developed complications out of which only 2 had major ones. Extreme care is required in FESS because of paranasal sinuses proximity to vital structures. Complications if identified early can be managed adequately

neglected case of foreign body bronchus

Majority of foreign bodies of tracheobroncial tree are found in children between the ages of 1-3 years. Foreign bodies once aspirated in air passages can lodge in larynx, trachea or bronchi but most commonly in right main bronchus. The course of disease, once the foreign body has lodged in air passages, depends upon the character of foreign body and its length of stay. Undiagnosed foreign body leads to complications such as recurrent pneumonia, bronchiectasis, atelactasis and even death. We are hereby reporting a case of neglected foreign body which remained in right main bronchus undiagnosed for 5 months. Patient was suffering from recurrent attacks of coughing, haemoptysis and breathlessness especially during sleep

Incidence of cholesteatoma in chronic suppurative otitis media, atticoantral disease

Background: Atticoantral variety of chronic suppurative otitis media also referred to as unsafe is a deep-seated inflammation of middle ear cleft involving underlying bone and is usually associated with cholesteatoma. The purpose of study was to see the presence of cholesteatoma in clinically diagnosed cases of chronic suppurative otitis media atticoantral type

Material and Methods: Sixty cases of chronic suppurative otitis media clinically diagnosed as atticoantral variety were included in this study. These patients had attic, marginal or total perforation and foul smelly discharge of more than two years duration. Mastoid exploration was done and findings were recorded for cholesteatoma and non-cholesteatoma ears

Results: Cholesteatoma was found in 45 patients [75%], and only granulations were seen in 15 patients [25%]

Conclusion: Patients having longstanding foul smelling discharge and attic perforation always have underlying cholesteatoma or granulation tissue

Pathogens involved and antibiotic sensitivity pattern of isolates in community acquired common bacterial skin infections presenting in a tertiary care hospital

To determine the bacterial pathogens involved and their antibiotic sensitivity pattern in commonly encountered community acquired superficial skin infections and determine the appropriate empirical antibiotics for such conditions. A descriptive study. A descriptive study was carried out at Department of Dermatology and Pathology Combined Military Hospital Peshawar from 1st Oct 2005 to 31st Jan 2006. Swabs from skin lesions of a total of 117 patients suffering from common primary pyodermas were collected for microbiological analysis. These were stained with gram stain and conventional biochemical and serological tests were performed for identification of different isolates after their culture followed by their antibiotic sensitivity testing with standard antibiotic discs using modified Kirby Bauer disk diffusion method as per National Committee for Clinical Laboratory Standards recommendations. A total of 117 cases were studied with age range from 6months to 60 years [median age 12 years]. Male and female ratio was 1.8:1. There were 12 cases, which did not reveal any growth. Among 69 positive cases of impetigo/ecthyma, in 14 [20.3%] Staphylococcus aureus was isolated; in 26 [37.7%] Streptococcus pyogenes; in 27 [39.1%] both Staphylococcus aureus and Streptococcus pyogenes and in only 2 cases [2.9%] both Staphylococcus aureus and Pseudomonas aerugenosa together were isolated. Among 26 positive folliculitis/furunculosis cases, in 19 [73.1%] Staphylococcus aureus; in 4 [15.4%] Streptococcus pyogenes and in 3 [11.5%] both these organisms were isolated. Among 10 positive cases of cellulitis, in 3 each [30%] Streptococcus pyogenes and Staphylococcus aureus alone and in 4 [40%] both these organisms together were isolated. In 105 positive cases, 72 Staphylococcus aureus and 67 Streptococcus pyogenes were isolated. In 34 cases both these organisms were isolated together. All Staphylococci were found resistant to all b lactamase labile penicillins and there were 6 [8.3%] staphylococci resistant to oxacillin [MRSA]. However, all Staphylococci other than MRSA were susceptible to b lactamase stable penicillins like Amoxicillin-Clavulanic acid. All the MRSA were susceptible to vancomycin and fusidic acid and some of them were susceptible to gentamicin, amikacin, ciprofloxacin, chloramphenicol, doxycycline and clindamycin. All streptococci were susceptible to penicillins, fusidic acid and vancomycin. Almost half of them were resistant to erythromycin and most to doxycycline. First choice for an empirical treatment of community acquired superficial skin infections should be b lactamase resistant penicillins like Amoxicillin-Clavulanic acid. In patients allergic to penicillin, lincosamides [Lincomycin and Clindamycin] or fusidic acid can be a systemic and/or topical alternative

Ossicular changes in chronic suppurative otitis media, atticoantral disease

Chronic suppurative otitis media is chronic inflammation of the middle ear characterized by ear discharge and hearing impairment. This hearing impairment is usually due to ossicular involvement and tympanic membrane perforation. The purpose of this study is to document the pattern of ossicular damage in chronic suppurative otitis media atticoantral type Sixty patients with chronic suppurative otitis media atticoantral type were selected and admitted in ENT-I Department. Ears were examined under microscope. Mastoid exploration was done in all cases. Per-operative ossicular chain status was noted. The sequence of damage to ossicles was documented and compared with other studies. Our conclusion was that in 57[95%] patients the ossicular chain was found damaged. Whole of the ossicular chain was involved in majority of the cases 25[42%]. Incus was the most vulnerable bone and was found necrosed in 20% isolated cases and in 46% along with other ossicles

Cutaneous vasculitis

Vasculitides constitute a heterogeneous group of inflammatory disorders involving walls of blood vessels of integument and viscera. Depending on the size of blood vessels affected and organs involved, different subsets can vary in clinical presentation, histopathology, laboratory markers, treatment and prognosis. The present review focuses on recapitulating the vasculitides involving skin in a tabulated form

Interaction of antifolates with enzymes of the de novo purine pathway in human CCRF-CEM leukaemia cells: Monitored using HPLC linked with UV and radioactive detectors

Estimation of the precursors of nucleic acids in the presence of anti-leukaemic developing agents is a valid approach towards seeking a cure for leukaemia. Therefore, in this study the focus was on primary inhibition of the early de novo purine pathway. A radiolabel protocol for optimum hot intermediates of the de novo purine biosynthesis produced in human leukaemia cells CCRF-CEM was developed in this study. According to recent earlier finding in this lab, methotrexate [MTX] and lome-trexol [LTX] showed blockage potency for the de novo synthesis ofpurines due to an accumulation of intracellular 5-phospho-ribosyl-1-pyrophosphate [PRPP]. Therefore, in the current study, the mechanism of action of these two antifolates on de novo purine biosynthesis in human CCRF-CEM leukaemia cells [labelled various metabolites with 14C-glycine] was elucidated by using high-pressure liquid chromatographic [HPLC] technique for separation of acid soluble metabolites and quantified using an ultraviolet detector along with a radioactivity monitor. Total level of nucleotides indicated that both MTX and LTX induced depletion ofATP, ADP, AMP, GTP, GMP, sAMP, IMP and NAD, while GDP was decreased by 63% of the control level only in the MTX [0.1 microM] treated sample. A level of nucleotides synthesized in the de novo purine biosynthesis indicated that both antifolates [0.1 microM] also induced depletion ofATP, ADP, AMP, GTP, GDP, IMP and NAD in treated human leukaemia cells. All mono-, di- and triphosphates of N-formylglycinamide ribotide [FGAR] were also decreased by MTX and LTX in the presence ofazaserine [25 microM]. These results support the finding that two tested antifolates in the present study induced primary inhibition of the early de novo purine pathway in human leukaemia cells CCRF-CEM, i.e. the inhibition of amido phosphoribosyl-transfemse [APRT] catalyses the first committed step of the de novo purine pathway

new, simple and economical approach to analyze the inhibition kinetics of acetylcholinesterase using tolserine

Tolserine is a novel and highly selective synthetic inhibitor of the enzyme acetylcholinesterase [AchE] that is being developed towards clinical trials for the treatment of Alzheimer's disease. We recently characterized its enzyme kinetics for inhibiting purified human erythrocyte AChE by utilizing classical methodology. In the current study, we describe a new approach to analyze its mechanism of inhibition of AChE. This was undertaken with purified human erythrocyte AChE using low to higher substrate concentrations in both the absence and presence of dual tolserine concentrations. The optical density of the generated reaction-product then was monitored during the initial reaction time after addition of each concentration of substrate. Thereafter, the new kinetic parameters [Kneidc, Kesic, Kslxx, Krss, Kslm and Kslx] were calculated from the resulting experimental data. These kinetic constants will hopefully aid our understanding of the mechanism of inhibittion and kinetic analysis of a variety of critical physiological enzymes by a wide assortment of inhibitors in vitro, during health, aging and disease. A low Kneidc value of 2.31 nM for tolserine indicates that it is a highly potent inhibitor of human erythrocyte AChE, which supports previous reports of its unusually high potency for inhibiting AChE both in vitro and in vivo, as compared to its structural analogues, physostigmine and phenserine

Post-tonsillectomy haemorrhage in a tertiary care teaching hospital

Post-tonsillectomy haemorrhage is a surgeon's nightmare. Despite all precautions some cases will always continue to pose a challenge to the ENT surgeons. This is a report of the authors experience in 600 consecutive tonsillectomies performed with the dissection method, over a period of 3 years. The incidence of haemorrhage was 1%, divisible into the two categories of reactionary haemorrhage in 0.33% and secondary haemorrhage in 0.67% of the case. The variables in this case control study comprised of the surgical experience, age and gender of the patients, use of absorabable sutures and postoperative antibiotic