Nylon 6-cellulose composite hosted in a hypodermic needle: Biofluid extraction and analysis by ambient mass spectrometry in a single device

This study proposes a hypodermic needle(HN)as a sorbent holder and an electrospray(ESI)emitter,thus combining extraction and analysis in a single device.A novel nylon 6-cellulose(N6-Cel)composite sorbent is proposed to extract methadone from oral fluid samples.The cellulosic substrate provides the composite with high porosity,permitting the flow-through of the sample,while the polyamide contributes to the extraction of the analyte.The low price of the devices(considering the holder and the sorbent)contributes to the affordability of the method,and their small size allows easy transportation,opening the door to on-site extractions.Under the optimum conditions,the analyte can be determined by high-resolution ambient ionization mass spectrometry at a limit of detection(LOD)as low as 0.3 mg/L and precision(expressed as relative standard deviation,RSD)better than 9.3%.The trueness,expressed as relative recovery(RR),ranged from 90%to 109%.As high-resolution mass spectrometers are not available in many laboratories,the method was also adapted to low-resolution spectrometers.In this sense,the direct infusion of the eluates in a triple quadrupole-mass spectrometry provided an LOD of 2.2 mg/L.The RSD was better than 5.3%,and the RR ranged from 96%to 121%.

Postaurical injection is a systemic delivery supported by symmetric distribution of Gd-DOTA in both the ipsilateral and contralateral ears

Postaurical injection of therapeutics was recently applied in clinical practice to treat inner ear diseases based on supposed existence of a direct channel from the postaurical area to the inner ear. Doubting on the associated reports and aiming to provide evidence on the inner ear uptake mechanism, the present study tracked the dynamic distribution of gadolinium-tetra-azacyclo-dodecane-tetra-acetic acid (Gd-DOTA) in rat inner ears after postaurical injection using MRI. A targeted tympanic medial wall delivery was utilized as control. The results showed that, at the early time points after postaurical injection, Gd-DOTA distributed mainly in tissues surrounding the bulla, temporal bone and skull and neck space. In the inner ear, there was gradual uptake of Gd-DOTA on both the ipsilateral and contralateral sides with equal signal intensities. There was no sign of direct channel carrying the agent from the postaurical area to the inner ear. Targeted tympanic medial wall delivery induced significantly greater uptake of Gd-DOTA in the inner ear than did postaurical injection. At 30 min post-administration, targeted tympanic medial wall delivery yielded 4.6-folds higher signal intensity than did postaurical injection. The total dose of Gd-DOTA delivered by the targeted tympanic medial wall approach was only 0.1% of that delivered by postaurical injection. In conclusion, postaurical injection is a systemic administration, which is similar to hypodermic injection, rather than a focal delivery method. By contraries, targeted tympanic medial wall delivery induces fast and abundant uptake of Gd-DOTA in the ipsilateral inner ear without significant distribution in unwanted areas.

Prevalence of Three Shrimp Viruses in Zhejiang Province in 2008

White spot syndrome virus (WSSV),Taura syndrome virus (TSV) and Infectious hypodermal and haematopoietic necrosis virus (IHHNV) are three shrimp viruses responsible for major pandemics affecting the shrimp farming industry. Shrimps samples were collected from 12 farms in Zhejiang province,China,in 2008 and analyzed by PCR to determine the prevalence of these viruses. From the 12 sampling locations,8 farms were positive for WSSV,8 for IHHNV and 6 for both WSSV and IHHNV. An average percentage of 57.4% of shrimp individuals were infected with WSSV,while 49.2% were infected with IHHNV. A high prevalence of co-infection with WSSV and IHHNV among samples was detected from the following samples:Bingjiang (93.3%),liuao (66.7%),Jianshan (46.7%) and Xianxiang (46.7%). No samples exhibited evidence of infection with TSV in collected samples. This study provides comprehensive information of the prevalence of three shrimp viruses in Zhejiang and may be helpful for disease prevention control in this region.

Adipose Derived Stem Cells and Growth Factors Applied on Hair Transplantation. Follow-Up of Clinical Outcome

Different studies show the need of immature adipose cell to induce the proliferation of bulge stem cells in order to kick off the anagen phase of hair cycle. Furthermore, the adipose derived stem cell, adipose progenitors, and growth factors secreted by mature adipocytes can help the wound healing and the vascular neogenesis. Nowadays, it is not known any protocol of tissue regeneration applied to hair transplantation, especially if aimed to the reconstruction of the main vascular network for the engraftment of transplanted hair and the healing process. The aim of the work is to investigate how the application of autologous cellular suspension obtained by Rigenera system, mechanical fragmentation procedure which allows to obtain a physiological saline solution consisting of a heterogeneous pool of cells rich in adipose derived mesenchymalstem cells and growth factors, helps the wound healing and engraftment of the transplanted hair. During hair restoration surgery, the adipose tissue recovered from the discard of follicular slicing, was processed using the Rigenera system. The obtained cell suspension was applied in the area of hair transplantation, increasing the natural background of adipocyte lineage and raising the amount of growth factors. In addition, the cellular suspension was applied to the suture on the occipital region. The cell population was characterized by FACS. The monthly evaluation of hair transplantation follow-up with photos and the patient’s impressions demonstrates that there is a faster healing of the micro-wound and a continuous growth of the transplanted hair even two months after the procedure, with a shortening of the dormant phase. In conclusion, this new approach aims to integrate regenerative medicine and hair restoration surgery in order to improve the outcome for the patient. It would be wonderful to continue this research to elaborate on the molecular cause behind this satisfying clinical.

Construction and expression of retroviral vector pLEGFP-N1-TERT in preparation of seed cells for skin tissue engineering

Objective:To construct the retroviral vector pLEGFP-N1-telomerase reverse transcriptase(TERT)and to investigate the expression of TERT in neonatal mouse bypodermal cells.Methods:The polymerase chain reaction(PCR)-amplified TERT gene was inserted into plasmid pLEGFPN1.The positive clone was identified by restriction enzyme digestion and sequencing,then was transfected into packaging cells to produce retrovirus particles.Neonatal mouse hypodermal cells were infected with the virus to generate a stable cell line.The TERT mRNA expression level,telomerase activity,and enhanced green fluorescent protein(EGFP)expression level were analyzed.Results:Retroviral vector pLEGFP-N1-TERT was constructed successfully,and a stable cell line of neonatal mouse hypodermal cells expressing EGFP was established.Western blot and immunohistochemical assay showed that the expression level of TERT was significantly elevated in the neonatal mouse hypodermal cells.Conclusions:A high titer of retrovirus pLEGFP-N1-TERT mediates high-level expression of the exogenous TERT gene in the neonatal mouse hypodermal cells.This protocol has potential applications for skin tissue engineering and cell transplantation therapy.

Erythema Nodosum Manifestation Post COVID-19 Vaccine: A Case Report

Erythema nodosum (EN) is a delayed hypersensitivity response that may be triggered by a range of conditions, including infections and vaccines. Rare cases of EN caused by COVID-19 were recently reported but none due to COVID-19 vaccines were documented. We report here a case of EN occurring after COVID-19 vaccination. Patient presented with painful nodular lesions of all 4 limbs, evolving for one month. These lesions appeared 48 h after the second dose of COVID-19 vaccination. The patient reported no recent infectious episodes. The physical examination found numerous, erythematous dermohypodermatitis knots with no palpable adenopathy. Some were regressive according to biligenesis shades. Biology and radiology findings eliminated other common causes of this dermatosis. The skin biopsy was done and suggested EN. The final diagnosis was post COVID-19 vaccine EN. Patient received a symptomatic treatment and had a slight improvement of the lesions 10 days after diagnosis. Physicians should be aware of the side effects of the vaccine including skin manifestations, especially since more people are bound to be vaccinated.