Hammerhead ribozyme-based U-insertion and deletion RNA editing assays for multiplexing in HTS applications.

Untranslatable mitochondrial transcripts in kinetoplastids are decrypted post-transcriptionally through an RNA editing process that entails uridine insertion/deletion. This unique stepwise process is mediated by the editosome, a multiprotein complex that is a validated drug target of considerable interest in addressing the unmet medical needs for kinetoplastid diseases. With that objective, several in vitro RNA editing assays have been developed, albeit with limited success in discovering potent inhibitors. This manuscript describes the development of three hammerhead ribozyme (HHR) FRET reporter-based RNA editing assays for precleaved deletion, insertion, and ligation assays that bypass the rate-limiting endonucleolytic cleavage step, providing information on U-deletion, U-insertion, and ligation activities. These assays exhibit higher editing efficiencies in shorter incubation times while requiring significantly less purified editosome and 10,000-fold less ATP than the previously published full round of in vitro RNA editing assay. Moreover, modifications in the reporter ribozyme sequence enable the feasibility of multiplexing a ribozyme-based insertion/deletion editing (RIDE) assay that simultaneously surveils U-insertion and deletion editing suitable for HTS. These assays can be used to find novel chemical compounds with chemotherapeutic applications or as probes for studying the editosome machinery.

Modified Treatment Of Shallow Fornix For Better Retention Of Ocular Prosthesis.

Background: The purpose of this study is to share the experience of a modified surgical technique for treating shallow inferior fornix to enhance the retention of cosmetic ocular prosthesis in a tertiary care hospital. It was a Quasi experimental study, carried out at Khalid Eye Hospital, Karachi, Pakistan from January to December 2018. Methods: This study included twenty-five patients between the ages of 20-40 years belonging to either gender having an anophthalmic socket with shallow inferior fornices unable to retain cosmetic ocular prosthesis. Three pieces of Silicone tube were placed deep within the inferior fornix with the help of 2-0 Vicryl suture, taking bite of the inferior orbital rim periosteum. Main outcome measure was deepening of the inferior fornix with better retention of the ocular prosthesis. All the patients were followed up for a period of one year after the surgery. Study dynamics were briefed to all the patients and study approval obtained from the hospital ethical review committee. Results: This study included a total of twenty-five patients of both genders. Mean age was 29.6±6.53 years. By the end of the follow up period, twenty-four (96%) out of the twenty-five patients were satisfied with a proper retention of ocular prosthesis within the fornix along with complete lid closure. One (4%) patient required revision surgery because of extrusion of the silicone tube piece after one month. Mean follow up period was 358.5±16.76 days. Conclusion: This modified technique does not require expensive consumables and proved to be effective in terms of cosmetic and functional outcome.

Frequency of Causes of Portal Hypertension in Children.

Introduction The most common etiology of portal hypertension (PH) in children is obstruction at the presinusoidal or sinusoidal level. In addition, portal vein thrombosis (PVT) and biliary atresia are the most prevalent extrahepatic causes. This study aims to evaluate all the possible etiologies leading to PH in the pediatric population and provide the most common cause associated with this condition along with the age group most frequently affected by it. Material and Methods From January 2018 to December 2020, a cross-sectional study was carried out in tertiary care hospitals in Pakistan. A total of 100 children, both male and female, aged one month to 15 years and diagnosed with PH, were enrolled for the evaluation for the causes of PH. The Statistical Package for the Social Sciences (SPSS), version 20, was used to analyze the data. Results The mean age of enrolled participants was 9.01 ± 2.81 years. It was found that PVT (63%) was the most common cause of PH, followed by liver cirrhosis (19%) and biliary atresia (18%). Age of more than eight years was significantly associated with PVT (p-value: 0.007). Conclusion In children, PH may be caused by a wide range of etiologies. It is imperative to understand the underlying etiologies contributing to PH for proper guidance and management, prevention, and overall outcomes.

An ensemble based approach using a combination of clustering and classification algorithms to enhance customer churn prediction in telecom industry.

Mobile communication has become a dominant medium of communication over the past two decades. New technologies and competitors are emerging rapidly and churn prediction has become a great concern for telecom companies. A customer churn prediction model can provide the accurate identification of potential churners so that a retention solution may be provided to them. The proposed churn prediction model is a hybrid model that is based on a combination of clustering and classification algorithms using an ensemble. First, different clustering algorithms (i.e. K-means, K-medoids, X-means and random clustering) were evaluated individually on two churn prediction datasets. Then hybrid models were introduced by combining the clusters with seven different classification algorithms individually and then evaluations were performed using ensembles. The proposed research was evaluated on two different benchmark telecom data sets obtained from GitHub and Bigml platforms. The analysis of results indicated that the proposed model attained the highest prediction accuracy of 94.7% on the GitHub dataset and 92.43% on the Bigml dataset. State of the art comparison was also performed using the proposed model. The proposed model performed significantly better than state of the art churn prediction models.

A Rare Case of Multidrug-Resistant Tuberculosis Affecting the Pleura.

The majority of cases with tuberculous pleuritis have negative acid-fast bacilli (AFB) on smear microscopy, making the diagnosis difficult. This case report is based on the successful diagnosis and management of an extra-pulmonary (EP) multidrug-resistant tuberculosis (MDR-TB) patient with a history of lymphoma. Initial tests revealed a right-sided pleural effusion and thickening of the pleura. The closed pleural biopsy, pleural fluid histopathology, culture, and drug sensitivity testing (DST) report revealed Mycobacterium tuberculosis with isoniazid and rifampicin resistance. Based on the DST report, the patient was labeled as a case of MDR-TB and successfully managed with an individualized drug-resistant TB (DR-TB) regimen. With initial negative microscopy and GeneXpert MTB/RIF (Sunnyvale, CA: Cepheid Inc.) reports, this case demonstrated that DR-TB could exist even in the absence of risk factors. Furthermore, it also unveils the importance of line probe assays (LPAs) and culture in identifying MDR-TB. Lymphocytic/exudative pleural effusions and pleural biopsy specimens should be subjected early on to investigations like Xpert/MTB RIF, cultures, and genotypic DST to timely diagnose and treat DR-TB.

Clinicopathological Analysis and Surgical Outcome of Eyelid Malignancies: A Study of 332 Cases.

BACKGROUND: Eyelid tumours are common in our ophthalmic practice. Malignancy cases account only for one-fourth of all eyelid tumours. The most aggressive eyelid malignancy is sebaceous gland carcinoma, but its occurrences are rare in western countries. We found sebaceous gland carcinoma is as common as basal cell carcinoma in our clinical practices. Hence, it is essential to build awareness about the more aggressive eyelid malignancies to reduce morbidity and mortality. AIM: To assess the relative frequency of eyelid malignancies in the Bangladesh population, state their clinical features and outcome of management strategies and build awareness about the more aggressive eyelid malignancies to reduce morbidity and mortality. METHODS: This was a retrospective case series study of 332 patients in Bangladesh. This study analyzed all the recorded data of the histologically proven primary eyelid malignancies and followed them up for at least six months from 2014 to 2019 (6 years). All patients were managed by surgical excision with tumor-free margins verified on histopathology, either the frozen section or excision biopsy with 2-3 mm microscopic view of normal tissue followed by eyelid reconstruction. Computer-based statistical software SPSS was used for the analysis, and an appropriate test of significance (chi-square) was used for the statistical analysis. RESULTS: Sebaceous gland carcinoma (SGC) was the highest in occurrence, at 42%, followed by 38% basal cell carcinoma (BCC), 18% squamous cell carcinoma (SqCC), and 02% malignant melanoma (MM). The mean age at presentation of SGC, BCC, SqCC, and MM were 57.41 years, 62.56 years, 64.73 years, and 59.28 years, respectively. Female (59%) was slightly more preponderance over the male (41%) for SGC than other malignancies. Pigmentation was associated with malignant melanoma (100%) and BCC (81%). Statistically, a significant difference was found between eyelid malignancies, including location, size, pigmentation, recurrence, and invasiveness. The recurrence rate was low lower in the patients who underwent frozen section biopsy (3%) for margin clearance than those who underwent excision biopsy (21.5%) in the follow-up time. Conjunctival map biopsy (8%) was performed as an essential tool for excluding the pagetoid spread of SGC. A new reconstruction method named triangular-shaped musculocutaneous tail flap was performed in 33 (11%) patients to reconstruct the moderate eyelid defect following local resection. CONCLUSION: Sebaceous gland carcinoma (SGC) was the highest occurrence found to be the highest occurrence among all eyelid malignancies in Bangladesh. SGC is more aggressive and the recurrence rate was higher than BCC and SqCC.

Characterization of the variable lymphocyte receptor B genes of a freshwater nonparasitic lamprey species, Lampetra hubbsi.

Agnathans possess a convergent adaptive immune system in comparison to that of jawed vertebrates. In lieu of immunoglobulins, agnathans deploy variable lymphocyte receptors (VLRs), single chain protein effector molecules consisting of leucine rich repeat modules. Foundational work for this discovery utilized the parasitic sea lamprey, Petromyzon marinus. However, for several reasons, it is desirable to employ a local species for further studies of lamprey immunity. A disjunct freshwater species from the Kings River of California, Lampetra hubbsi, was evaluated for this purpose. Validation that its adaptive immune system was analogous to that of P. marinus entailed detailed examination of its immune tissue organization and of its VLRB cDNA transcripts. The VLRB molecules showed high degrees of homology with P. marinus VLRB. Furthermore, hemato-lymphopoietic tissue expression of VLRB protein was confirmed. We conclude that L. hubbsi should be a viable alternative for studying the lamprey adaptive immune system and for generation of monoclonal antibodies.

Preexisting opioid use disorder is associated with poor outcomes in hospitalized acute pancreatitis patients.

INTRODUCTION: Although opioids are widely used for pain management in acute pancreatitis, the impact of opioid use disorder (OUD) on outcomes in patients with acute pancreatitis remains unknown. In the current study, we aimed to evaluate the impact of the OUD on outcomes in patients hospitalized with acute pancreatitis and delineate the trends associated with OUD and acute pancreatitis using a nationally representative sample. METHODS: This is a retrospective cohort study of patients with acute pancreatitis using the combined releases of the year 2005-2014 of the National (Nationwide) Inpatient Sample (NIS) database. Patients over the age of 18 years with a principal diagnosis of acute pancreatitis were divided into cohorts of patients with opioid use disorders and those without. The primary measured outcome was in-hospital mortality and secondary outcomes were healthcare utilization measures, including length of stay (LOS) and hospitalization costs. RESULTS: A total of 2 593 831 hospitalizations of acute pancreatitis were included; of which, 37 849 (1.46%) had a secondary diagnosis of OUD. Total acute pancreatitis-related hospitalizations increased from 237 882 in 2005 to 274 006 in 2014. At the same time prevalence of OUD in acute pancreatitis patients also increased from 1 to 2.1%. Patients with OUD had significantly increased mortality as compared to patients without OUD (aOR: 1.4; P < 0.001). At the same time, acute pancreatitis patients with OUD were associated with 1.3 days longer LOS as compared to other acute pancreatitis patients (P < 0.001]. The mean adjusted difference in total hospitalization costs was $2353 (P < 0.001). CONCLUSION: OUD is associated with a significant increase in LOS, healthcare utilization cost and in-hospital mortality in patients admitted for acute pancreatitis. Therefore, clinicians should exercise caution in prescribing opioid medications to this high-risk patient population and other modalities such as nonopioid pain medications should be tried as alternatives to opioid analgesics.

Early Chest Re-Exploration for Excessive Bleeding in Post Cardiac Surgery Patients: Does It Matter?

Introduction Re-explorations after open-heart surgery are often required if the patient is bleeding or shows features of cardiovascular instability and does not improve with conservative measures. Our study aims to determine whether timely re-exploration of patients who are bleeding has an impact on the morbidity and mortality of the patients. Methods A retrospective analysis of 75 patients that underwent open-heart surgery and subsequently underwent chest re-exploration for excessive bleeding between March 2018 and March 2020. Patients who were reopened post-op for indications other than excessive bleeding were excluded. Results A total number of cases were 700, out of which 75 (9.3%) patients were reopened, as compared to the literature, which shows worldwide 2-11% being reopened. Post-operative drain output was 1000ml to 1500ml in 47 (62.7%) and more than 1500ml in 28 (37.3%) patients before they were reopened. In 67 (89.3%) patients, three to five units of blood were transfused, and in eight (10.7%) patients, more than five units of blood were transfused. We believe our mortality in the reopened patients was low, because of timely intervention and early re-exploration, and is probably the reason why our figures land in a higher range (2-11%) of reopened cases (9.3%). Reopening time was less than five hours in 49 (65.3%) patients and less than 10 hours in 26 (34.7%) patients in our study. We tried to minimize the loss of blood and re-explored the patients before they lose excessive blood, the average time for reopening in our study was less than 10 hours. The average intensive care unit (ICU) stay was 4.2 days (range three to six days). Wound infections were reported in one of three patients. There was no mortality in these patients. Surgical site of bleeding was identified in 54 (72%) patients and no particular site was found in 21 (28%) patients. Suggesting that it is common to have a surgical bleeder than coagulopathy induced bleeding in post-cardiac surgery patients Conclusions We believe our low mortality (0%) is due to early reopening in patients who are bleeding excessively after cardiac surgery.

Incidence and Causes of Reintubation Other Than Reopening of the Chest in Post-Cardiac Surgical Patients in a Tertiary Care Hospital.

Objective To determine the incidence of endotracheal reintubation, excluding surgical reopening, in post-cardiac surgical patients in a tertiary care hospital. Material and methods A retrospective descriptive analysis of 408 patients who underwent different cardiac surgeries during this period. Post-operative extubation was performed when patients fulfilled the preset criteria for extubation, which include consciousness (awake and aware), stable vital signs, acceptable arterial blood gases, acceptable respiratory mechanics, a maximum inspiratory force greater than 20-25 cm H2O, chest tube drainage less than 100 ml per hour, normal temperature and electrolytes. The total number of patients who were reintubated within 72 hours of extubation was noted. The criteria for reintubation included altered conscious level with Glasgow Coma Score (GCS) of less than 8, respiratory failure, unstable hemodynamics, and arrhythmias such as ventricular tachycardia (VT) and fibrillation. All of the information was collected retrospectively on a specifically prepared form. Data was entered and evaluated in Statistical Package for the Social Sciences. The research was piloted in the Cardiac Intensive Care Unit (CICU) of Northwest General Hospital and Research Center, Hayatabad, Peshawar from December 2018 to March 2020. Results Out of 408 patients who had cardiac surgeries, only nine (2.2%) were reintubated after initial extubation. The average time for which patients remained on the ventilator was 8 ± 2 hours. The reasons for reintubation were recorded. Among those reintubated, eight patients (88.88%) had undergone coronary artery bypass grafting (CABG) whereas one patient (11.11%) had undergone mitral valve replacement (MVR). In three (33.33%) patients, stroke (hemiplegia in two and global brain ischemia in one) with low GCS was the primary cause of reintubation. Arrhythmias - which included VT, ventricular fibrillation (VF), and supraventricular tachyarrhythmias (SVT) - were responsible for three (33.33%) cases of reintubation. Respiratory failure - with a partial pressure of oxygen in arterial blood less than 60 mmHg, along with tachypnea - was responsible for reintubation in two (22.22%) patients. In one (11.11%) patient who had MVR, cardiac arrest was the underlying reason; the cause of arrest could not be retrieved from the retrospective data. Notably, as a common variable, five (62.5%) out of the eight reintubated CABG patients had a poor left ventricular function.  Conclusion Causes of reintubation were primarily cardiac (arrhythmias) and neurological, followed by respiratory causes in our center. Patients with poor left ventricular function and diffuse coronary artery disease appear to have a higher incidence of reintubation which can lead to extended CICU and hospital stay, elevated mortality, and higher costs.

Outcome Of Recurrent Macular Hole Closure With Amniotic Membrane Plug.

BACKGROUND: The surgery of macular hole is a complex and intricate micro-surgery and chances of recurrence of macular hole are always high. Therefore, in order to provide a medium to close this hole, we carried out this research on subjects using amniotic membrane, in light of the studies being conducted around the world. This in turn led to benefitting the patients by improving their vision over time. PURPOSE: To assess the rate of recurrent macular hole closure with amniotic membrane plug. It was a Quasi-experimental study, conducted at Layton Rahmatullah Benevolent Trust (LRBT) Free Eye Hospital Karachi, from January 2019 to January 2020. METHODS: This study was conducted using 13 eyes of 13 patients with recurrent macular hole who underwent amniotic membrane plugging via pars plana approach. Outcomes measured were changes in best corrected visual acuity (BCVA) and change in hole size with the help of optical coherence tomography (OCT). RESULTS: Anatomic closure was attained in 100% of the cases whereas BCVA improved from 1.7±0.33 (6/300) to 0.9±0.15 (6/48). CONCLUSIONS: The use of AM is a functional method for management of large RMH.

The Experience Of Intralesional Bleomycin For Orbital Vascular Anomalies.

BACKGROUND: Orbital vascular anomalies are congenital and pose a challenge. They require various treatment modalities which are often unsuccessful. The use of intralesional Bleomycin has gained popularity in recent times as a scar less procedure when other treatment modalities are not effective. PURPOSE: To share the experience of using intra lesional Bleomycin in orbital vascular anomalies in a Tertiary care hospital. METHODS: This quasi-experimental study was carried out at Layton Rahmatullah Benevolent Trust (LRBT) Teaching Tertiary care hospital, Karachi from 1st June to 31st December 2018. It included fifteen patients of various orbital vascular anomalies treated with injection of intra lesional sclerosing (Bleomycin) agent. Patients were given multiple injections and the effects of sclerosing agent on orbital vascular anomalies were observed. RESULTS: Results were excellent in terms of regression of vascular anomalies. CONCLUSIONS: Intralesional sclerosing agent (Bleomycin) has an outstanding role in vascular tumour.

To assess the role of cataract extraction in glaucoma management by its intraocular pressure lowering effect.

OBJECTIVE: To measure the mean change of intraocular pressure in glaucoma patients with cataract after uncomplicated phacoemulsification surgery with intraocular lens implanted in capsular bag. METHODS: The quasi-experimental study was conducted at the Ophthalmology Department of Pakistan Institute of Medical Sciences Shaheed Zulfiqar Ali Bhutto Medical University, Islamabad, Pakistan, from June 11 to December 10, 2018, and comprised patients who had uncomplicated cataract extraction by phacoemulsification with intraocular lens implant in the capsular bag in glaucomatous eyes of age 30-80 years. Visual acuity, intraocular pressure, slit lamp examination, fundoscopy, visual fields, details about topical medication and relevant history were recorded not more than 5 days before cataract extraction. Intraocular pressure was recorded using Goldman's applanation tonometer one day before surgery, and post-surgery 1 month and 3 months. Data was analysed using SPSS 20. RESULTS: Of the 40 patients, 19(47.50%) were males and 21(52.50%) were females. The overall mean age was 52.23±9.44 years. Mean pre-operation intraocular pressure was 20.42±1.69mmHg, while at 1 month post-surgery it was 18.55±0.90mmHg and at 3 months it was 17.03±1.19mmHg (p=0.0001). CONCLUSIONS: There was a significant change in intraocular pressure readings in glaucoma patients with cataract after uncomplicated phacoemulsification surgery with intraocular lens implanted in capsular bag.

Pulmonary Hypertension as a Predictor of Early Outcomes of Mitral Valve Replacement: A Study in Rheumatic Heart Disease Patients.

Aim Mitral valve pathology in rheumatic heart disease patients is a common cause of secondary pulmonary hypertension (PH). Our aim was to evaluate pulmonary hypertension severity as a predictor of in-hospital mortality and early complications following mitral valve replacement. Methods A retrospective review of rheumatic heart disease patients who underwent mitral valve replacement between January 2017 and August 2020 was performed. Systolic pulmonary artery pressure (sPAP) was used to classify patients as no PH (<35 mmHg), mild PH (35-44 mmHg), moderate PH (45-59 mmHg) or severe PH (>60 mmHg). Patients subjected to additional cardiac procedures (such as aortic valve replacement and coronary artery bypass grafting) were excluded from the study sample. Results The study group was composed of 159 patients (mean age: 40; 73 male, 86 female) categorized as no PH (n = 32; 20.1%), mild PH (n = 14; 8.8%), moderate PH (n = 65, 40.9%) and severe PH (n = 48, 30.2%) groups. Patient demographic data and preoperative comorbidities were comparable among the four groups. Use of intraoperative and postoperative blood products was similar in all the groups. Severe PH patients had similar in-hospital mortality (4.2%; p = 0.74) as in groups with lesser degrees of pulmonary hypertension. Likewise, increasing severity of pulmonary hypertension did not confer any significant increase in early postoperative complications, namely prolonged ICU stay (10.4%; p = 0.41), prolonged ventilation (2.1%; p = 0.70), reintubation (4.2%; p = 0.90), reopening for bleeding tamponade (6.3%; p = 0.39), new-onset renal failure (6.3%; p = 0.91), postoperative stroke (4.2%; p = 0.52) or prolonged length of stay (mean: 5.6 + 2.8 days; p = 0.49). Conclusions Increasing severity of pulmonary hypertension does not appear to have a significant impact on in-hospital mortality or early postoperative outcomes of patients undergoing mitral valve replacement.

A Network Meta-Analysis Comparing Osteoporotic Fracture among Different Direct Oral Anticoagulants and Vitamin K Antagonists in Patients with Atrial Fibrillation

Background@#There are limited studies comparing the risk of osteoporosis and fractures between different direct oral anticoagulants (DOACs) and vitamin K antagonists (VKA) in non-valvular atrial fibrillation (AF). Using a network meta-analysis (NMA), we compared osteoporotic fractures among 5 different treatment arms, viz. dabigatran, rivaroxaban, apixaban, edoxaban, and VKA. @*Methods@#Ten studies, including 5 randomized control trials and 5 population-based studies, with a total of 321,844 patients (148,751 and 173,093 in the VKA and DOAC group, respectively) with a median follow-up of 2 years, were included. A Bayesian random-effects NMA model comparing fractures among the treatment arms was performed using MetInsight V3. Sensitivity analysis excluded studies with the highest residual deviances from the NMA model. @*Results@#The mean age of the patients was 70 years. The meta-analysis favored DOACs over VKA with significantly lower osteoporotic fracture (odds ratio [OR], 0.77; 95% credible interval [CrI], 0.70-0.86). The NMA demonstrated that fractures were significantly lower with apixaban compared with dabigatran (OR, 0.64; 95% CrI, 0.44-0.95); however, fractures were statistically similar between apixaban and rivaroxaban (OR, 0.84; 95% CrI, 0.58-1.24) and dabigatran and rivaroxaban (OR, 1.32; 95% CrI, 0.90-1.87). Based on the Bayesian model of NMA, the probability of osteoporotic fracture was highest with VKA and lowest with apixaban, followed by rivaroxaban, edoxaban, and dabigatran. @*Conclusions@#The decision to prescribe anticoagulants in elderly patients with AF should be made not only based on thrombotic and bleeding risks but also on the risk of osteoporotic fracture; these factors should be considered and incorporated in contemporary cardiology practice.

A Network Meta-Analysis Comparing Osteoporotic Fracture among Different Direct Oral Anticoagulants and Vitamin K Antagonists in Patients with Atrial Fibrillation

Background@#There are limited studies comparing the risk of osteoporosis and fractures between different direct oral anticoagulants (DOACs) and vitamin K antagonists (VKA) in non-valvular atrial fibrillation (AF). Using a network meta-analysis (NMA), we compared osteoporotic fractures among 5 different treatment arms, viz. dabigatran, rivaroxaban, apixaban, edoxaban, and VKA. @*Methods@#Ten studies, including 5 randomized control trials and 5 population-based studies, with a total of 321,844 patients (148,751 and 173,093 in the VKA and DOAC group, respectively) with a median follow-up of 2 years, were included. A Bayesian random-effects NMA model comparing fractures among the treatment arms was performed using MetInsight V3. Sensitivity analysis excluded studies with the highest residual deviances from the NMA model. @*Results@#The mean age of the patients was 70 years. The meta-analysis favored DOACs over VKA with significantly lower osteoporotic fracture (odds ratio [OR], 0.77; 95% credible interval [CrI], 0.70-0.86). The NMA demonstrated that fractures were significantly lower with apixaban compared with dabigatran (OR, 0.64; 95% CrI, 0.44-0.95); however, fractures were statistically similar between apixaban and rivaroxaban (OR, 0.84; 95% CrI, 0.58-1.24) and dabigatran and rivaroxaban (OR, 1.32; 95% CrI, 0.90-1.87). Based on the Bayesian model of NMA, the probability of osteoporotic fracture was highest with VKA and lowest with apixaban, followed by rivaroxaban, edoxaban, and dabigatran. @*Conclusions@#The decision to prescribe anticoagulants in elderly patients with AF should be made not only based on thrombotic and bleeding risks but also on the risk of osteoporotic fracture; these factors should be considered and incorporated in contemporary cardiology practice.

Evidence of chitin in the ampullae of Lorenzini of chondrichthyan fishes.

We previously reported that the polysaccharide chitin, a key component of arthropod exoskeletons and fungal cell walls, is endogenously produced by fishes and amphibians in spite of the widely held view that it was not synthesized by vertebrates [1]. Genes encoding chitin synthase enzymes were found in the genomes of a number of fishes and amphibians and shown to be correspondingly expressed at the sites where chitin was localized [1,2]. In this report, we present evidence suggesting that chitin is prevalent within the specialized electrosensory organs of cartilaginous fishes (Chondrichthyes). These organs, the Ampullae of Lorenzini (AoL), are widely distributed and comprise a series of gel-filled canals emanating from pores in the skin (Figure 1A). The canals extend into bulbous structures called alveoli that contain sensory cells capable of detecting subtle changes in electric fields (Figure 1B) [3,4]. The findings described here extend the number of vertebrate taxa where endogenous chitin production has been detected and raise questions regarding chitin's potential function in chondrichthyan fishes and other aquatic vertebrates.

Clinical outcomes and safety profile of Tenecteplase in wake-up stroke.

BACKGROUND: Tenecteplase has probably pharmacological and clinical advantages in the treatment of acute ischemic stroke. There are lacking data about safety and efficacy of tenecteplase in wake-up stroke (WUPS). AIMS: To investigate safety and efficacy of tenecteplase compared to alteplase in WUPS patients included in NOR-TEST. METHODS: WUPS patients in NOR-TEST were included in the study based on DWI-FLAIR mismatch. Included patients randomly assigned (1:1) to receive intravenous tenecteplase 0.4 mg/kg (to a maximum of 40 mg) or alteplase 0.9 mg/kg (to a maximum of 90 mg). Neurological improvement was defined as 1) favorable functional outcome at 90 days modified Rankin Scale (mRS) of 0 or 1 and 2) neurological improvement measured with the National Institutes of Health Stroke Scale (NIHSS) of 4 points within 24 hours as compared to admission NIHSS or NIHSS 0 at 24 hours. RESULTS: Of 1100 patients from 13 stroke centers included in NOR-TEST, 45 were WUPS patients. Of these, 5 patients were stroke mimics and excluded. Of the remaining 40 patients (3.6%), 24 were treated with alteplase (60%). There was no difference in the number of patients achieving a good clinical outcome (mRS 0-1) in either treatment group. Patients treated with tenecteplase showed a better early neurological improvement (87.5% vs 54.2%, P = 0.027). No ICH was detected on MRI/CT 24-28 hours after thrombolysis. CONCLUSIONS: In WUPS patients treated in NOR-TEST, there was no difference in clinical outcomes at 90 days and no ICH events or deaths were observed in either alteplase- or tenecteplase-treated patients. Clinical Trial Registration-URL: https://www.clinicaltrials.gov. Unique identifier: NCT01949948.

Generation of Lamprey Monoclonal Antibodies (Lampribodies) Using the Phage Display System.

The variable lymphocyte receptors (VLRs) consist of leucine rich repeats (LRRs) and comprise the humoral antibodies produced by lampreys and hagfishes. The diversity of the molecules is generated by stepwise genomic rearrangements of LRR cassettes dispersed throughout the VLRB locus. Previously, target-specific monovalent VLRB antibodies were isolated from sea lamprey larvae after immunization with model antigens. Further, the cloned VLR cDNAs from activated lamprey leukocytes were transfected into human cell lines or yeast to select best binders. Here, we expand on the overall utility of the VLRB technology by introducing it into a filamentous phage display system. We first tested the efficacy of isolating phage into which known VLRB molecules were cloned after a series of dilutions. These experiments showed that targeted VLRB clones could easily be recovered even after extensive dilutions (1 to 109). We further utilized the system to isolate target-specific "lampribodies" from phage display libraries from immunized animals and observed an amplification of binders with relative high affinities by competitive binding. The lampribodies can be individually purified and ostensibly utilized for applications for which conventional monoclonal antibodies are employed.

A comprehensive review on tyrosinase inhibitors.

Tyrosinase is a multi-copper enzyme which is widely distributed in different organisms and plays an important role in the melanogenesis and enzymatic browning. Therefore, its inhibitors can be attractive in cosmetics and medicinal industries as depigmentation agents and also in food and agriculture industries as antibrowning compounds. For this purpose, many natural, semi-synthetic and synthetic inhibitors have been developed by different screening methods to date. This review has focused on the tyrosinase inhibitors discovered from all sources and biochemically characterised in the last four decades.