Metabolomic profiling reveals altered phenylalanine metabolism in Parkinson's disease in an Egyptian cohort.

Introduction: Parkinson's disease (PD) is the most common motor neurodegenerative disease worldwide. Given the complexity of PD etiology and the different metabolic derangements correlated to the disease, metabolomics profiling of patients is a helpful tool to identify patho-mechanistic pathways for the disease development. Dopamine metabolism has been the target of several previous studies, of which some have reported lower phenylalanine and tyrosine levels in PD patients compared to controls. Methods: In this study, we have collected plasma from 27 PD patients, 18 reference controls, and 8 high-risk controls to perform a metabolomic study using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). Results: Our findings revealed higher intensities of trans-cinnamate, a phenylalanine metabolite, in patients compared to reference controls. Thus, we hypothesize that phenylalanine metabolism has been shifted to produce trans-cinnamate via L-phenylalanine ammonia lyase (PAL), instead of producing tyrosine, a dopamine precursor, via phenylalanine hydroxylase (PAH). Discussion: Given that these metabolites are precursors to several other metabolic pathways, the intensities of many metabolites such as dopamine, norepinephrine, and 3-hydroxyanthranilic acid, which connects phenylalanine metabolism to that of tryptophan, have been altered. Consequently, and in respect to Metabolic Control Analysis (MCA) theory, the levels of tryptophan metabolites have also been altered. Some of these metabolites are tryptamine, melatonin, and nicotinamide. Thus, we assume that these alterations could contribute to the dopaminergic, adrenergic, and serotonergic neurodegeneration that happen in the disease.

Extender Supplementation with Glutathione (GSH) and Taurine Improves In Vitro Sperm Quality and Antioxidant Status of New Zealand Rabbits during Chilled Storage for up to 72 hours.

This study assessed the influence of supplementing the rabbit semen extender with various concentrations of glutathione (GSH) and taurine at 24, 48, and 72 h postchilling at 5°C. Semen samples were collected from 20 New Zealand bucks, and ejaculates with standard color, motility (>85%), about 0.5 mL volume, and ∼400 × 106/mL concentration were used and diluted with extenders supplemented with 0.5, 1, and 2 mM of GSH and 1, 5, and 10 mM of taurine and chilled at 5°C. Nonsupplemented samples were used as a control. Sperm's progressive motility, acrosome reaction, and extracellular oxidative stress biomarkers such as MDA contents and GPx, SOD, and CAT concentrations and intracellular transcriptomic levels of SOD and CAT genes were assessed. GSH and taurine supplementation improved the sperm's kinetics by reducing cooling-associated stress, which was ascertained by lowering MDA concentration and increasing SOD, CAT, and GPx concentrations (P < 0.05). Increasing the levels of antioxidant enzymes in the extender was due to the increasing mRNA copies of the SOD and CAT genes (P < 0.05). Furthermore, GSH and taurine maintained the fructose levels in the extender and lowered the GPT levels, which implies sperm membrane stability is maintained through GSH and taurine supplementation. GSH and taurine supplementation to the extender had protective influences on the in vitro rabbit semen quality during chilled storage for up to 72 h, which were remarkable with increasing supplementation dose and cooling time at 5°C.

A Core-Shell Au@TiO2 and Multi-Walled Carbon Nanotube-Based Sensor for the Electroanalytical Determination of H2O2 in Human Blood Serum and Saliva.

A hydrogen peroxide (H2O2) sensor was developed based on core-shell gold@titanium dioxide nanoparticles and multi-walled carbon nanotubes modified glassy carbon electrode (Au@TiO2/MWCNTs/GCE). Core-shell Au@TiO2 material was prepared and characterized using a scanning electron microscopy and energy dispersive X-ray analysis (SEM/EDX), transmission electron microscopy (TEM), atomic force microscopy (AFM), Raman spectroscopy, X-ray diffraction (XRD) and Zeta-potential analyzer. The proposed sensor (Au@TiO2/MWCNTs/GCE) was investigated electrochemically using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The analytical performance of the sensor was evaluated towards H2O2 using differential pulse voltammetry (DPV). The proposed sensor exhibited excellent stability and sensitivity with a linear concentration range from 5 to 200 µM (R2 = 0.9973) and 200 to 6000 µM (R2 = 0.9994), and a limit of detection (LOD) of 1.4 µM achieved under physiological pH conditions. The practicality of the proposed sensor was further tested by measuring H2O2 in human serum and saliva samples. The observed response and recovery results demonstrate its potential for real-world H2O2 monitoring. Additionally, the proposed sensor and detection strategy can offer potential prospects in electrochemical sensors development, indicative oxidative stress monitoring, clinical diagnostics, general cancer biomarker measurements, paper bleaching, etc.

Outcomes of the Pedicled Medial Sural Artery Perforator Flap for Soft Tissue Reconstruction Around the Knee: When to Use It and How to Look After It.

Introduction The medial gastrocnemius flap is an established workhorse flap to cover proximal leg defects due to its reliability and simplicity to raise. However, it has the disadvantage of being bulky, requiring a skin graft for coverage, and is associated with loss of muscle power. The pedicled medial sural artery perforator (MSAP) flap has gained popularity as a reconstructive alternative for defects of the lower extremities. We present a case series of pedicled MSAP flaps for reconstructing defects around the knee as an alternative to the medial gastrocnemius flap. Materials and Methods A consecutive series of patients with proximal leg defects following trauma, osteomyelitis, burns, and chronic wounds were included. A hand-held Doppler was used to map out the MSAPs. Defects were reconstructed using pedicled MSAP flaps, preserving the nerve supply to the gastrocnemius muscle. Patient outcomes were recorded, including their Enneking scores postreconstruction. Results A total of 10 pedicled flaps was performed to reconstruct defects around the knee joint between October 2017 and November 2018. All the patients were discharged 1 week postoperatively, and rehabilitation consisted of graduated flexion in a knee brace by means of controlled passive mobilization. Three out of the ten patients developed complications: one patient developed flap congestion, one developed epidermolysis of the tip of the flap, and the other patient had partial necrosis of the skin paddle. The average Enneking score was 29 out of 35. Conclusion The pedicled MSAP flap is a good reconstructive option for proximal leg defects as it is associated with lower donor site morbidity and provides an aesthetically pleasing reconstruction.

Mycobacterium szulgai Infection after Carpal Tunnel Release Requiring Extensive Debridement and ALT Flap Reconstruction.

Nontuberculous mycobacteria (NTM) refer to all mycobacteria species not associated with the Mycobacterium tuberculosis complex. NTM have been shown to cause superficial and deep tissue infections of the hand, the vast majority of which are attributed to Mycobacterium marinum . There have been only eight cases of Mycobacterium szulgai -associated infections of the hand described in the English literature, none requiring surgical reconstruction. We describe a severe case of M. szulgai -associated carpal tunnel infection reported in an immunocompetent patient requiring extensive soft tissue debridement and reconstruction. This case illustrates the importance of a multidisciplinary team approach with microbiology specialists when managing severe soft tissue infections of the hand.

Extracorporeal Membrane Oxygenation (ECMO)-A Lifesaving Technology. Review and Single-center Experience.

OBJECTIVE: Extracorporeal membrane oxygenation is used to bypass the cardiopulmonary system in a severe heart or/and lung failure, mainly in intractable conditions where all other therapy options fail or are unfeasible. Extracorporeal membrane oxygenation (ECMO) is a well-established therapeutic option in such circumstances for neonatal, pediatric, and adult patients. Managing a patient with ECMO requires dedicated and specific management. The importance and necessity of this essential technology in life-threatening cardio-respiratory rescue prompted Rambam Health Care Campus to implement it and make it available as a service to the population in northern Israel. This article includes a brief review of extracorporeal life support and a report of our single-center experience since the establishment of the service. METHODS: The ECMO unit was established in 2014 under the responsibility of the Cardiac Surgery Department. The ECMO service was initiated by a well-planned program with consideration of all aspects including economics, education and training, the specialist team and equipment needed, strategies for medication, and ethical challenges. RESULTS: Between February 2014 and May 2018, 65 patients were treated with ECMO; 43 patients received veno-arterial ECMO for cardiac support (66%), while 22 received veno-venous ECMO for respiratory support (34%). The in-hospital mortality was 56%. CONCLUSIONS: Extracorporeal membrane oxygenation is an effective therapy that is constantly growing in use and provides a therapy that can replace previous options. To establish such a service requires a planned program and concerted effort. Our single-center experience presented a good learning curve and showed the feasibility as well as the efficacy of the ECMO procedure in life-threatening conditions.

Antioxidant and developmental capacity of retinol on the in vitro culture of rabbit embryos.

SummaryOxidative stress is a major cause of defective embryo development during in vitro culture. Retinoids are recognized as non-enzymatic antioxidants and may have an important role in the regulation of cell differentiation and vertebrate development. However, there are not enough reports discussing the antioxidant and developmental capacity of retinoids, including retinol (RT), on the in vitro development of embryos recovered from livestock animals, particularly in rabbit species. Therefore, morula embryos obtained from nulliparous Red Baladi rabbit does were cultured for 48 h in TCM199 medium in the absence of RT (control group) or in the presence of RT at concentrations of 10, 100 and 1000 nM. The developmental capacity to the hatched blastocyst stage, the antioxidant biomarker assay and the expression of several selected genes were analyzed in each RT group. The data show that RT significantly (P<0.001) promoted the embryo hatchability rate at the concentration of 1000 nM to 69.44% versus 29.71% for the control. The activity of malondialdehyde (MDA) level was significantly (P<0.05) lower in the RT groups than in the control group, while the total antioxidant capacity (TAC), superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities were significantly (P<0.05) higher following treatment with RT. Furthermore, RT treatment considerably upregulated the relative expression of gap junction protein alpha 1 (GJA1), POU class 5 homeobox 1 (POU5F1) and superoxide dismutase 1 (SOD1) genes compared with the control group. The current study highlights the potential effects of RT as antioxidant in the culture medium on the in vitro development of rabbit embryos.

Contraceptive Implant-Related Acute Ulnar Neuropathy: Prompt Diagnosis, Early Referral, and Management Are Key.

Objective: Subdermal contraceptive implants are a well-established method of contraception. Aside from common side effects such as irregular menstrual periods, headaches, and weight gain, rarer complications related to their insertion and removal have been reported. These include traumatic peripheral neuropathy after procedures to remove nonpalpable implants. Only 2 cases of ulnar neuropathy after insertion of a contraceptive implant have been described in the literature, one which resolved spontaneously and another in which postoperative recovery was not described. We report a case of acute ulnar nerve neuropathy in a patient postinsertion of a contraceptive implant who achieved symptom resolution after prompt referral and treatment at a specialist plastic surgery center. Methods: A 22-year-old, right-hand-dominant woman presented 1 day postinsertion of a contraceptive implant (Implanon) in her left arm with paresthesia along the ulnar distribution of her hand and forearm, as well as shooting pain on palpating the course of the ulnar nerve. Ultrasonography found the implant to be lying in the subfascial plane of the inner arm. Results: The implant was found lying in the perineurium of the ulnar nerve, causing ulnar nerve neuropathy. A review 3 months after removal of the implant showed near-complete resolution of her symptoms. Conclusions: Complications related to implantable contraceptives may lead to significant morbidity. Appropriate training for health care professionals administering the devices is essential, as well as early referral to a specialist center to prevent permanent damage.

Peripheral Vascular Disease: The Beneficial Effect of Exercise in Peripheral Vascular Diseases Based on Clinical Trials.

Intermittent claudication (IC) due to peripheral artery diseases (PAD) is one of the disabling disease that can affect quality of life (QOL) and functional status of capacity. It is characterized by cramping pain which develops with exercise and eliminated by rest secondary to decrease blood flow to the muscles. The annual incidence rate is increased with age. Exercise rehabilitation has a great impact in improving the functional capacity and prevent the functional disability. The available evidences from current studies have showed that exercise therapy is considered the primary treatment in PAD, which in consequently improves the QOL. In this chapter we will illustrate the current available evidences which support exercise benefit and outcomes in PAD with IC.

DNA biosensors based on gold nanoparticles-modified graphene oxide for the detection of breast cancer biomarkers for early diagnosis.

Two different DNA (ERBB2c and CD24c) modified gold nanoparticles and graphene oxide loaded on glassy carbon electrodes were prepared for early detection of breast cancer markers by electrochemical detection of HER2. Comparative study of ERBB2c and CD24c for the detection was carried out. A "sandwich-type" detection strategy was employed in this electrochemical DNA biosensor and its response was measured by amperometric detection. The electrochemical signal enhancement achieved via gold nanoparticles and grapheme oxide system allowed for sensitive detection of the breast cancer biomarker ERBB2 and the control marker CD24. The modified graphene oxide was characterised using Raman spectroscopy, UV-visible spectroscopy, Fourier transform infrared spectroscopy transmission electron microscopy, scanning electron microscopy and energy-dispersive X-ray spectroscopy. The various steps involved in the modification of a glassy carbon electrode with graphene oxide, gold nanoparticles and DNA probes, target and reporter probe were electrochemically characterised using cyclic voltammetry and electrochemical impedance spectroscopy. Using amperometric detection of a horse radish peroxidase label, detection limits of 0.16nM and 0.23nM were obtained with sensitivity 378nA/nM and 219nA/nM for ERBB2 andCD24 respectively.

Ultrasensitive, rapid and inexpensive detection of DNA using paper based lateral flow assay.

Sensitive, specific, rapid, inexpensive and easy-to-use nucleic acid tests for use at the point-of-need are critical for the emerging field of personalised medicine for which companion diagnostics are essential, as well as for application in low resource settings. Here we report on the development of a point-of-care nucleic acid lateral flow test for the direct detection of isothermally amplified DNA. The recombinase polymerase amplification method is modified slightly to use tailed primers, resulting in an amplicon with a duplex flanked by two single stranded DNA tails. This tailed amplicon facilitates detection via hybridisation to a surface immobilised oligonucleotide capture probe and a gold nanoparticle labelled reporter probe. A detection limit of 1 × 10-11 M (190 amol), equivalent to 8.67 × 105 copies of DNA was achieved, with the entire assay, both amplification and detection, being completed in less than 15 minutes at a constant temperature of 37 °C. The use of the tailed primers obviates the need for hapten labelling and consequent use of capture and reporter antibodies, whilst also avoiding the need for any post-amplification processing for the generation of single stranded DNA, thus presenting an assay that can facilely find application at the point of need.

Antioxidant Capacity of Melatonin on Preimplantation Development of Fresh and Vitrified Rabbit Embryos: Morphological and Molecular Aspects.

Embryo cryopreservation remains an important technique to enhance the reconstitution and distribution of animal populations with high genetic merit. One of the major detrimental factors to this technique is the damage caused by oxidative stress. Melatonin is widely known as an antioxidant with multi-faceted ways to counteract the oxidative stress. In this paper, we investigated the role of melatonin in protecting rabbit embryos during preimplantation development from the potential harmful effects of oxidative stress induced by in vitro culture or vitrification. Rabbit embryos at morula stages were cultured for 2 hr with 0 or 10-3 M melatonin (C or M groups). Embryos of each group were either transferred to fresh culture media (CF and MF groups) or vitrified/devitrified (CV and MV groups), then cultured in vitro for 48 hr until the blastocyst stage. The culture media were used to measure the activity of antioxidant enzymes: glutathione-s-transferase (GST) and superoxide dismutase (SOD), as well as the levels of two oxidative substrates: lipid peroxidation (LPO) and nitric oxide (NO). The blastocysts from each group were used to measure the expression of developmental-related genes (GJA1, POU5F1 and Nanog) and oxidative-stress-response-related genes (NFE2L2, SOD1 and GPX1). The data showed that melatonin promoted significantly (P<0.05) the blastocyst rate by 17% and 12% in MF and MV groups compared to their controls (CF and CV groups). The GST and SOD activity significantly increased by the treatment of melatonin in fresh or vitrified embryos, while the levels of LPO and NO decreased (P<0.05). Additionally, melatonin considerably stimulated the relative expression of GJA1, NFE2L2 and SOD1 genes in MF and MV embryos compared to CF group. Furthermore, melatonin significantly ameliorated the reduction of POU5F1 and GPX1 expression induced by vitrification. The results obtained from the current investigation provide new and clear molecular aspects regarding the mechanisms by which melatonin promotes development of both fresh and vitrified rabbit embryos.

In vitro development rate of preimplantation rabbit embryos cultured with different levels of melatonin.

This study aimed to investigate the effect of melatonin supplementation at different levels in culture medium on embryo development in rabbits. Embryos of 2-4 cells, 8-16 cells and morula stages were recovered from nulliparous Red Baladi rabbit does by laparotomy technique 24, 48 and 72 h post-insemination, respectively. Normal embryos from each stage were cultured to hatched blastocyst stages in either control culture medium (TCM-199 + 20% fetal bovine serum) or control supplemented with melatonin at 10(-3) M, 10(-6) M or 10(-9) M. No effect of melatonin was found on development of embryos recovered at 24 h post-insemination. The high level of melatonin at 10(-3) M adversely affected the in vitro development rates of embryos recovered at 48 h post-insemination (52 versus 86, 87 and 80% blastocyst rate; 28 versus 66, 78 and 59% hatchability rate for 10(-3) M versus 10(-9) M, 10(-6) M and control, respectively, P< 0.05). At the morula stage, melatonin at 10-3 M significantly increased the in vitro development of embryos (92% for 10(-3) M versus 76% for control, P < 0.05), while the hatchability rate of these embryos was not improved by melatonin (16-30% versus 52% for melatonin groups versus control, P < 0.05). Results show that a moderate level of melatonin (10(-6) M) may improve the development and hatchability rates of preimplantation rabbit embryos. The addition of melatonin at a 10-3 M concentration enhances the development of rabbit morulae but may negatively affect the development of earlier embryos. More studies are needed to optimize the use of melatonin in in vitro embryo culture in rabbits.

Oviductal and endometrial mRNA expression of implantation candidate biomarkers during early pregnancy in rabbit.

Prenatal losses are a complex problem. Pregnancy requires orchestrated communication between the embryo and the uterus that includes secretions from the embryo to signal pregnancy recognition and secretion and remodelling from the uterine epithelium. Most of these losses are characterized by asynchronization between embryo and uterus. To better understand possible causes, an analysis was conducted of gene expression of a set of transcripts related to maternal recognition and establishment of rabbit pregnancy (uteroglobin, SCGB1A1; integrin α1, ITGA1; interferon-γ, IFNG; vascular endothelial growth factor, VEGF) in oviduct and uterine tissue at 16, 72 or 144 h post-ovulation and insemination. In the oviduct tissue, a significant decrease in the level of SCGB1A1 mRNA expression was observed from 144 h post-ovulation. In the case of ITGA1, the transcript abundance was initially lower, but mRNA expression increased significantly at 72 and 144 h post-ovulation. For IFNG, a huge decrease was observed from 16 to 72 h post-ovulation. Finally, no significant differences were observed in the VEGF transcript. For the endometrium, the results showed a significant decline in the level of SCGB1A1 mRNA expression from 16 to 144 h post-ovulation induction. The highest levels of ITGA1 transcript were detected at 144 h, followed by the 16 h group and lower at 72 h post-ovulation. For IFNG there were no significant differences among post-ovulation induction times. Finally, it was possible to observe that VEGF mRNA abundance was present at low levels at 16 h post-ovulation and remained low at 72 h, but increased at 144 h. The functional significance of these observations may provide new insights into the maternal role in prenatal losses.

Ocular injury requiring hospitalisation in the south east of Ireland: 2001-2007.

AIM: To investigate whether recent socio-demographic changes and recent health and safety measures have impacted on the trends of ocular trauma in the South East of Ireland. METHODS: We retrospectively reviewed all cases of ocular trauma admitted to our department between October 2001 and September 2007, and the following data were retrieved: demographic details; mechanism of injury and nature of injury. RESULTS: During the study period, 517 patients were admitted with ocular trauma. Work-related and home-related activities were the commonest causes of admission, and accounted for 160 (31.8%) and 145 (28.4%) cases, respectively. In 2006/2007, and following the influx of migrant workers from the 10 new EU accession states (EUAS), the incidence of hospitalised ocular injuries per 100,000 was 89 in persons from the EUAS versus 18 in those of Irish origin, P < or = 0.0001. After adding the offence of not wearing a seat belt to the traffic penalty point system in Ireland, the proportion of road traffic accident (RTA)-related ocular injuries dropped significantly from 6.7% to 2.4%, P=0.03. CONCLUSION: The inclusion of the offence of not wearing a seat belt in the traffic penalty point system may have contributed to the significantly lower proportion of hospitalised ocular injuries attributable to RTAs. Also, the demographic profile of patients admitted because of ocular trauma has changed over the last 6 years, reflected in an increasing proportion of these injuries in persons from the EUAS. These data will inform healthcare providers, and those involved in developing health and safety guidelines for the workplace.

Risk factors, microbiological findings, and clinical outcomes in cases of microbial keratitis admitted to a tertiary referral center in ireland.

AIM: To identify the risk factors for, and to report the microbiological findings and clinical outcomes of, severe microbial keratitis (MK). METHODS: This was a retrospective study of all cases of presumed MK admitted to a tertiary referral center over a 2-year period (September 2001 to August 2003). Data recorded included demographic data, details relating to possible risk factors, results of microbiological studies, clinical findings at presentation, and clinical and visual outcomes. RESULTS: Ninety patients were admitted with a diagnosis of presumed MK during the study period. The mean age of patients was 45 +/- 32 years, and the male to female ratio was 47:43 (52.2%:47.7%). Predisposing risk factors for MK included contact lens wear (37; 41.1%), anterior segment disease (19; 21.1%), ocular trauma (13; 14.4%), systemic disease (5; 5.6%), and previous ocular surgery (1; 1.1%). Cultured organisms included gram-negative bacteria (17; 51.5%), gram-positive bacteria (11, 33.3%), acanthamoeba (2; 6.1%), and fungi (1; 3%). Visual acuity improved significantly after treatment [mean best-corrected visual acuity (+/-standard deviation) at presentation: 0.76 (+/-0.11); mean best-corrected visual acuity at last follow-up: 0.24 (+/-0.07); P < 0.001]. Secondary surgical procedures were required in 18 (20%) cases, and these included punctal cautery (1; 1.1%), tissue glue repair of corneal perforation (2; 2.2%), tarsorrhaphy (9; 9.9%), Botulinum toxin-induced ptosis (1; 1.1%), penetrating keratoplasty (3; 3.3%), and evisceration (2; 2.2%). CONCLUSIONS: Contact lens wear remains a significant risk factor for severe MK. MK remains a threat to vision and to the eye, but the majority of cases respond to prompt and appropriate antimicrobial therapy.