“It’s more than milk, it’s mental health”: a case of online human milk sharing

Background Milk sharing is not a new concept and occurs today via regulated human milk banks and unregulated online milk sharing groups. Exploring and understanding how, and why, mothers use these peers to peer milk sharing groups, is a vehicle to understanding how breastfeeding mothers can be tangibly supported online, adding to the literature on peer milk sharing, from a recipient’s perspective. This research presents a single case example of an online breastfeeding support group use, through one mother’s experiencing of seeking human donor milk. Method This is a qualitative, exploratory study observing the attitudes, thoughts, and feelings of one mother who is seeking human donor milk through online groups. A single key case was identified, and the participant was asked to document thoughts and feelings as she searched for milk online. A telephone interview was conducted after two months, and the online page activity from the Human Milk for Human Babies Facebook group was captured for the week following the interview. The results were presented in a chronological and linear analytical approach adopting pattern matching. Results ‘Abbi’ is a mother who has Polycystic Ovary Syndrome and subsequent low milk supply and sought donor breastmilk online. Online support groups introduced her to donor milk sharing, which not only supported her breastfeeding but supported her own mental health. Abbi talks of the need to build a trusting relationship with her donor, due to the lack of regulation, and the positive impact it had for her and ‘Lucas’, her baby. Conclusion Considering milk sharing groups simply as tangible online support ignores the complexities around Abbi’s decision to use human donor milk. Peer milk sharing online is an option for mothers, but it is surrounded by stigma amongst other mothers, professionals, and even within pro breastfeeding support groups.

Descemet Membrane Endothelial Keratoplasty: NGS Metagenomics-Optimized Tissue Preparation and Surgical Technique. Doctoral Thesis

Title: Next-generation sequencing for the detection of microorganisms present in human donor corneal preservation medium. Aim: To detect the presence of microorganisms in the storage media of human donor corneas using next-generation sequencing method. Methods: Seven samples from organ culture (OC) group (Cornea Max, Eurobio, Les Ulis, France) with one control (sterile media without any cornea) and seven samples from hypothermic storage group (Cornea Cold, Eurobio) with one control were used for this study. The corneas were placed in the respective storage media for 14 days before collecting the samples. Storage media (2 mL) from each sample were collected in RNAase-free tubes and shipped for ribosomal RNA sequencing of 16S and 18S. Simultaneously, another 1 mL of media sample was used for conventional diagnostic method (CDM) using Bactec instruments. Results: In both, OC and hypothermic storage and control samples, the most abundant genera were Pseudomonas, Comamonas, Stenotrophomonas, Alcanivorax, Brevundimonas and Nitrobacter. Acidovorax, Acetobacter and Hydrogenophilus were detected mostly in the hypothermic storage group. The most abundant fungal pathogen detected belonged to the genus Malassezia, which was found in both the storage conditions. CDM was negative for microorganisms in all the samples. Conclusion: Metagenomics provides full taxonomic profiling of the detected genomic material of the organisms and thus has the potential to deliver a much wider microbiological diagnostic approach than CDM. The costs and turnaround time need to be reduced, and; the detection of viable organisms would help this technology to be introduced into routine clinical practice.

Descemet Membrane Endothelial Keratoplasty: NGS Metagenomics-Optimized Tissue Preparation and Surgical Technique. Summary of the Doctoral Thesis

Title: Next-generation sequencing for the detection of microorganisms present in human donor corneal preservation medium. Aim: To detect the presence of microorganisms in the storage media of human donor corneas using next-generation sequencing method. Methods: Seven samples from organ culture (OC) group (Cornea Max, Eurobio, Les Ulis, France) with one control (sterile media without any cornea) and seven samples from hypothermic storage group (Cornea Cold, Eurobio) with one control were used for this study. The corneas were placed in the respective storage media for 14 days before collecting the samples. Storage media (2 mL) from each sample were collected in RNAase-free tubes and shipped for ribosomal RNA sequencing of 16S and 18S. Simultaneously, another 1 mL of media sample was used for conventional diagnostic method (CDM) using Bactec instruments. Results: In both, OC and hypothermic storage and control samples, the most abundant genera were Pseudomonas, Comamonas, Stenotrophomonas, Alcanivorax, Brevundimonas and Nitrobacter. Acidovorax, Acetobacter and Hydrogenophilus were detected mostly in the hypothermic storage group. The most abundant fungal pathogen detected belonged to the genus Malassezia, which was found in both the storage conditions. CDM was negative for microorganisms in all the samples. Conclusion: Metagenomics provides full taxonomic profiling of the detected genomic material of the organisms and thus has the potential to deliver a much wider microbiological diagnostic approach than CDM. The costs and turnaround time need to be reduced, and; the detection of viable organisms would help this technology to be introduced into routine clinical practice.

Ultrastructural Analysis of Rehydrated Human Donor Corneas After Air-Drying and Dissection by Femtosecond Laser

Purpose: To evaluate the efficiency of femtosecond laser (FSL) incision of rehydrated human donor corneas after air-drying and its effects on corneal structure.Methods: We compared the rehydrated and fresh-preserved corneas by microscopy following Victus-Tecnolas FSL treatment for straight-edge anterior lamellar keratoplasty (ALK). The corneas were dehydrated at room temperature under a laminar-flow hood.Results: To obtain the horizontal cut in rehydrated corneas, we increased the FSL pulse energy to 1.2 μJ from 0.80 μJ applied for the fresh corneas and obtained a clear-cut separation of the lamellar lenticule cap from the corneal bed. Light microscopy showed regular arrangement of stromal collagen lamellae, with spaces in between the fibers in the corneal stroma in the fresh and the rehydrated corneas, but the uppermost epithelial layers in the rehydrated corneas were lost. Transmission electron microscopy (TEM) revealed no signs of thermal or mechanical damage to the corneal structure. The epithelial basal membrane and Bowman's layer maintained their integrity. The epithelial basal layer and cells were separated by large spaces due to junction alteration in the rehydrated corneas. There were gaps between the lamellar layers in the stroma, especially in the rehydrated corneas. Keratocytes displayed normal structure in the fresh corneas but were devoid of microorganules in the rehydrated corneas. Minor irregularities were observed in the vertical incision and the horizontal stroma appeared smooth on scanning electron microscopy.Conclusion: The corneal stroma of rehydrated corneas maintained morphology and integrity, while corneal cellular components were generally altered. When corneas are intended for FSL-assisted ALK, effective stromal bed incision is best achieved at a laser power higher than that currently adopted for fresh corneas.

Neuroprotective role of human neuritin 1 on axotomized human eyes pressurized using the Translaminar Autonomous System

Purpose: Primary open-angle glaucoma is a leading cause of vision loss worldwide characterized by retinal ganglion cell death (RGC) and visual field loss. Glaucoma is most commonly associated with an increase in intraocular pressure (IOP), and current clinical management is limited to lowering intraocular pressure. We have previously shown that the secreted human protein neuritin 1 (NRN1) exhibits neuroprotection, regeneration, and preservation of RGC function after RGC axonal injury. We propose to overcome RGC degeneration by over-expressing NRN1 in our novel ex-vivo perfused pressurized Translaminar Autonomous System (TAS). Methods: Human donor eyes were obtained from eye banks. To model glaucomatous conditions, dissected human posterior eye cups were cultured in the TAS model with IOP:ICP conditions of 16:12 mmHg over 6 days. Flow rate of perfusion medium to the interior and exterior (around optic nerve) of the eye cup (IOP and ICP, respectively) was controlled by automated pumps to induce the translaminar pressure gradient.We assessed survival of RGCs with and without hNRN1 treatment by TaqMan array analysis of apoptosis/inflammatory markers (retina). Expression of FN and COLIV was examined from conditioned medium of both groups by Western analysis. Results: We successfully maintained IOP: ICP differentials over 6 days. The RGCs degenerated faster in non-treated conditions compared to NRN1 treated eyes. In contrast to control, NRN1-treated group displayed differential expression of FN and COLIV. We observed dysregulated expression of GFAP, BAX and TLR4 expression between control and experimental conditions. Conclusion: We utilized our unique TAS model to recreate the human optic nerve head environment of elevated translaminar pressure for testing a unique neuroprotective and regenerative gene therapy. Using this approach, we hope to identify human Neuritin 1 as a therapeutic gene to save the visual neurons and develop a new treatment for glaucoma.

Killing efficacy of a new hypothermic corneal storage medium against the micro-organisms frequently found in human donor cornea intended for transplantation

ObjectiveTo study the in vitro killing efficacy of Kerasave (AL.CHI.MI.A Srl), a medium provided with amphotericin B tablet for hypothermic storage of human donor corneas, against relevant contaminants associated with postkeratoplasty infections.Methods and AnalysisThe antimicrobial activity of Kerasave was determined after 0, 3 and 14 days of incubation at 2°C–8°C, inoculating Kerasave and the control medium with 105–106 colony forming units (CFU) of Candida albicans (CA), Fusarium solani (FS), Aspergillus brasiliensis (AB), Staphylococcus aureus (SA), Enterococcus faecalis (EF), Bacillus subtilis spizizenii (BS), Pseudomonas aeruginosa (PA), Enterobacter cloacae (EC) and Klebsiella pneumoniae (KP). Log10 reductions at different time intervals were determined by assessing the number of viable CFU using the serial dilution plating technique.ResultsAfter 3 days, Kerasave induced the highest log10 decrease in the concentrations of KP, PA, CA and EC (5.37, 4.15, 2.97 and 2.67, respectively; all p<0.001). The log10 decreases of SA and EF were 2.27 and 2.11, respectively (all p<0.001). The lowest log10 decrease was observed in BS, AB and FS concentrations (0.25, 0.30 and 0.67, respectively; p<0.001 for BS and AB and p=0.004 for FS). After 14 days, the microbial count of CA, FS, SA, EF, PA and EC further decreased (p=0.006 for FS; p<0.001 for the others).ConclusionKerasave effectively reduced or kept unchanged the microbial concentration of almost all tested strains after 3 days. Thus, this novel medium represents a valuable tool to control the microbial contamination of human donor corneas during hypothermic storage for up to 14 days before transplantation.

Gram stain and addition of amphotericin B to improve the microbial safety of human donor corneas

To determine the effectiveness of two methods to improve the microbial safety of human corneas preserved in organ culture. We compared the number of positive preservation solutions of corneas in organ culture in which the initial short-term hypothermic corneal maintenance solution was supplemented with amphotericin B 2.5 µg/mL and the historical data of microbial test results (2015–2019). In addition, we appraised the efficacy of Gram stain to detect bacterial or fungal contamination in the organ culture solutions of corneas from at-risk donors compared to the culture tests of corneas from not-at-risk donors. Statistical analysis was performed using STATA and statistical significance set at p < 0.05. The number of positive culture tests after preservation was 15 (0.5%) in 2020 compared to a mean of 37 (1.2%) in the period 2015–2019 (p < 0.01), with 10 (1.0%) positive samples in the cohort of 998 corneas from at-risk donors and 5 (0.2%) in the 2046 corneas from not-at-risk donors (p < 0.01). All corneas from at-risk donors tested positive at Gram stain and the results were available 1–3 days before those of the conventional culture tests. Amphotericin B supplementation in the short-term maintenance solution markedly reduced the number of positive microbial tests after organ culture and the early detection of contaminants, including slow-growing microorganisms, by Gram stain before the standard culture results. This meant fewer corneas being discarded and a greater likelihood of preventing post-graft infections.

Utilization of Paragonix SherpaPak for human donor heart preservation

A heart transplant is the gold standard treatment for end stage heart failure. Preservation of the donor heart during its transfer from the hospital of the donor to that of the recipient has a significant impact on the outcome of the transplant procedure. Icebox storage is a conventional method utilized for this purpose that may not provide uniform cooling of the donor heart and does not allow monitoring of the temperature of the donor heart during preservation. The Paragonix SherpaPak Cardiac Transport System offers uniform cooling by suspending the donor heart in a preservation solution and provides continuous temperature monitoring.