Ice Cold Research Articles

Investigation of nanoparticles present in biochemic tissue salt Kalium phosphoricum in different potencies

BackgroundBiochemic tissue salts are defined as essential inorganic salts needed to maintain homoeostasis within living tissue. Kali phosphoricum is one of the twelve biochemic tissue salts. Nanomedicine is an expanding field, in which nanoparticles play an integral part and are known to exist in the natural world and can also be manufactured by scientists. This study sought to determine whether Kali phos included nanoparticles and whether may help to mitigate brain problems. Material and MethodTo investigate whether potassium phosphate nanoparticles were present in Kali phos, ultraviolet-visible spectroscopy and Fourier transform infrared spectroscopy were used. To analyze the biochemical parameters, the homogenate of brain was incubated with lactose powder (control group), different potencies of Kali phos and K2HPO4. 25% homogenate of normal brain tissue was made with the help of ice cold 0.02M Tris-Cl (pH 7.4) buffer. Different antioxidant enzymes and glycolytic enzyme activities were measured. ResultsIt may be concluded from this investigation, utilizing these chosen potencies, that the biochemic tissue salt Kali phos does contain nanoparticles of potassium phosphate, adding to the development of the nanomedicine field. Lower potency remedies were more effective in modulating the level of antioxidant and glycolytic enzymes. ConclusionAlthough there is clearly a body of scientific evidence, mainstream medicine claims that there is simply an inadequate amount of such evidence. These results may expand the range of use and applications of biochemic tissue salts, lay the groundwork for the creation of empirical homoeopathic evidence, and bring innovative research avenues.

Representasi Budaya Korupsi Dalam Film Ice Cold: Murder, Coffee and Jessica Wongso

ABSTRACT Corruption has become a common problem in Indonesia. Corruption in Indonesia has existed since the colonial era and continues, starting from even the smallest level. Even in 2022, the Indonesian NGO Wach Corruption found 579 cases, 1,396 suspects, and state losses of IDR 42,747 trillion, bribes of IDR 693 billion, illegal levies of IDR 11.6 billion, and money laundering of IDR 955 billion. Therefore, this research aims to find out how the culture of corruption is represented in the film Ice Cold: Murder, Coffee and Jessica Wongso. Researchers use a qualitative approach and use semiotic methods, especially John Fiske's codes. The subject used in this research is the film Ice Cold: Murder, Coffee and Jessica Wongso, while the object of this research is the representation of corruption culture in the film Ice Cold: Murder, Coffee and Jessica Wongso. Through the film Ice Cold: Murder, Coffee and Jessica Wongso, Reza Indragiri and Yudi Wibowo succeeded in showing that in Indonesia today the culture of corruption is still very common, such as the bribery case told by Reza Indragiri and Yudi Wibowo's opinion that in Indonesia today everything is about money, which is even Justice can also be bought even by bribing the person concerned.

Melukis Kembali Penggambaran Citra Gender dalam Media Massa

In Netflix’s 'Ice Cold,' which delves into the murder of Jessica by Mirna, she is often praised and forgiven for her beauty. The documentary even states that the killer is a handsome figure. Women seem to be forced to conform to the image of modern women, who should hold feminist paradigms. Digital television content, as one of the mass media platforms, often portrays women as limited to the role of 'subordinates.' The existence of women is frequently reduced solely to their physical dimensions. It is as if advertisements and television programs would feel empty without the presence of women celebrated as objects. This perspective eventually becomes ingrained in the minds of modern women, largely through the influence of mass media, while commercial demands and the race for the highest ratings are the primary reasons why women continue to be used as complementary objects in various media content.This research employs Roland Barthes' Semiotics to explore how religious education, the male gaze, mass media, and societal views interact in shaping the contemporary perception of the female body. Ironically, the excessive use of patriarchal media has made women feel more liberated to express themselves and actualize their potential in various aspects of life. In this context, elements of religious education play a crucial role, as some Abrahamic religious teachings promote gender hierarchies that can influence society's perception of women and result in gender inequality. The concept of the male gaze is also relevant in this analysis, as it refers to how the male perspective dominates the understanding and representation of women in media, often reducing women to objects of male sexual desire. This research can offer valuable insights into the interconnection of these perspectives and women's potential for increased self-expression within patriarchal-dominated environments.

Gradual rewarming with a hemoglobin-based oxygen carrier improves viability of donation after circulatory death in rat livers.

Donation after circulatory death (DCD) grafts are vital for increasing available donor organs. Gradual rewarming during machine perfusion has proven effective in mitigating reperfusion injury and enhancing graft quality. Limited data exist on artificial oxygen carriers as an effective solution to meet the increasing metabolic demand with temperature changes. The aim of the present study was to assess the efficacy and safety of utilizing a hemoglobin-based oxygen carrier (HBOC) during the gradual rewarming of DCD rat livers. Liver grafts were procured after 30 min of warm ischemia. The effect of 90 min of oxygenated rewarming perfusion from ice cold temperatures (4 °C) to 37 °C with HBOC after cold storage was evaluated and the results were compared with cold storage alone. Reperfusion at 37 °C was performed to assess the post-preservation recovery. Gradual rewarming with HBOC significantly enhanced recovery, demonstrated by markedly lower lactate levels and reduced vascular resistance compared to cold-stored liver grafts. Increased bile production in the HBOC group was noted, indicating improved liver function and bile synthesis capacity. Histological examination showed reduced cellular damage and better tissue preservation in the HBOC-treated livers compared to those subjected to cold storage alone. This study suggests the safety of using HBOC during rewarming perfusion of rat livers as no harmful effect was detected. Furthermore, the viability assessment indicated improvement in graft function.

Analisis Wacana pada Percakapan Film “Ice Cold: Murder, Coffee and Jessica Wongso” dalam Mempengaruhi Opini Publik

Public opinion can be formed or influenced through various media that contain it. Through documentary films, an event can be raised to be reviewed in a different form. One of them is a documentary film produced by Netflix with the title “Ice Cold: Murder, Coffee and Jessica Wongso”. The movie presents a collection of conversations and new facts that the public did not know before. The purpose of this research is to find out what forms of conversation in movies can shape and influence public opinion. This research uses the method of conversation analysis syntagmatic discourse analysis with the approach of perlocution speech act theory. The results of this study found 6 classifications of perlocutionary speech acts that are able to influence public opinion, namely anger, request, order, confusion, affirmation, annoyance

Public Response to the Jessica Kumalawongso Murder Case Against the Films Ice Cold Murder, Coffee and Jessica Wongso: A Sociological Analysis of Law

Public Response to the Jessica Kumalawongso Murder Case Against the Films Ice Cold Murder, Coffee and Jessica Wongso: A Sociological Analysis of Law

Cancer Stem Cell and Epithelial Mesenchymal Transition (EMT) in Canine Transmissible Venereal Tumour- A Preliminary Study

Background: Cancer stem cell (CSC) and Epithelial to Mesenchymal Transition (EMT) transforms conversion of normal cells in the tumor microenvironment into CSC which is the main cause of embryogenesis, carcinogenesis and metastasis. Methods: Nineteen canine patients with recurrent transmissible venereal tumor (TVT) and metastasis of inguinal lymph node were the subjects of present study. Preoperative work up and clinical staging involving radiography, ultrasonography, hematobiochemical analysis and histopathology or biopsy were done to confirm the type of cancerous tissue. Tumor tissues were collected in ice cold RPMI-1640 and phosphate buffer solution. Flow cytometry analysis was performed following established protocol from surgically excised tissue sample. Trypan blue dye exclusion test was done to quantify the live/dead cell count. CD24PE, CD44FITC, CD133APC and EpCAM antibody with conjugates were chosen markers for flow cytometry analysis. Result: Nineteen canine patients were diagnosed with Transmissible Venereal tumor (TVT) (common sexually transmitted disease in male dog) through biopsy of excised surgical tissue following Haematoxylin and Eosin staining protocol. Recurrence was seen even after adjuvant chemotherapy with Vincristine injection @ 0.25 mg/m2 at weekly interval for 5 weeks. Trypan blue dye exclusion test confirms 98% live cell count in transmissible venereal granuloma after digestion and incubation for flow cytometry. Flow cytometry analysis of the cells from the TVT showed surface biomarkers for EpCAM+ cells, EpCAM CD24+ cells, EpCAM CD44+ cells, EpCAM CD133+ indicating cells with CSC and EMT properties. Presence of CSC and EMT cells in this patient was responsible for chemoresistance, distant metastasis and recurrence. Identification of specific CSC in a malignant cancer will yields deeper insight into biological aspects of metastasis and formulation of targeted therapy in fighting with cancer.

The linkage between autumn Barents-Kara sea ice and European cold winter extremes

While the Arctic's accelerated warming and sea ice decline have been associated with Eurasian cooling, debates persist between those attributing this to sea ice retreat and those to internal variability. Our study examines the association between autumn sea ice variability over the Barents-Kara Seas and extreme cold winters in Europe. Using the observational data and composite analysis, we explore the interannual variability and the potential linkage between sea ice and atmospheric circulation patterns. It reveals a correlation with shifts toward a negative phase of North Atlantic Oscillation and more frequent episodes of the atmospheric blocking over Greenland and the North Atlantic. Furthermore, the negative phase of the North Atlantic Oscillation and enhanced blocking are closely related and mutually reinforcing, shaping the spatial distribution of cold anomalies over much of the European continent. Our results suggest a link between the unusual decrease in Barents-Kara Sea ice during autumn and the occurrence of intense European weather extremes in subsequent winter months, emphasizing the need for delving deeper into this relationship on monthly time scales to enhance our predictive capabilities for midlatitude extreme events.

Problematika Penegakan Hukum Kasus Pembunuhan Berencana di Indonesia: Studi Kasus Pembunuhan Berencana Kopi Sianida oleh Jessica Wongso terhadap Mirna Salihin

This research was conducted to look at the problems of law enforcement in Indonesia in the case of the premeditated murder of Wayan Mirna Salihin. Whether the judge's decision has provided justice that is as fair as possible to the victim and perpetrator, the evidence presented at the trial does not comply with procedures in accordance with the Criminal Code, the emergence of the films Ice Cold Murder, Coffee and Jessica Wongso has made the public less confident in the law in Indonesia. It is necessary to examine the role of the forensic autopsy in uncovering this case, the discovery of cyanide in Mirna's stomach, which is very strong evidence to prove that the cause of Mirna's death was poisoning by her friend Jessica Wongso. Research methods used used in this research is normative legal research, research using a problem approach, namely judicial case study by approaching the Mirna Salihin premeditated murder case, by examining the judge's decision which resulted in Jessica Wongso being convicted, by conducting this research uncovering the irregularities that occurred in This murder case creates a conflict of interest that cannot be resolved. Jessica Kumala Wongso has been convicted of violating Article 340 of the Criminal Code, namely premeditated murder. In the results of the examination, experts said that a complete autopsy must be carried out for human deaths due to poison. The expert concluded that the cause of WAYAN MIRNA SALIHIN's death was cyanide, especially since 0.2 mg/liter of cyanide was found in the stomach. The judge's decision regarding the premeditated murder of Mirna Salihin did not have any valid and strong evidence at all, starting from the testimony of witnesses who did not there was someone who saw Jessica put poison in the coffee that Mirna was drinking, then the expert's testimony in the examination was not perfect, one of which was that the autopsy did not comply with proper procedures, as well as evidence, such as CCTV which was not original and contrary to regulations. KAPOLRI No. 10 of 2009, as well as the defendant's statement that it never occurred to him to kill his own friend.

Multi-objective optimization of ice distribution uniformity and startup failure time for cold start of PEMFC catalyst layer based on CTnet and PSO-GA

The distribution of ice in the cathode catalyst layer (CCL) of a proton exchange membrane fuel cell (PEMFC) has a significant impact on cold start. A three-dimensional multiphase numerical model for fuel cell cold start was developed to investigate the first-order finite difference sensitivities of seven selected parameters, namely gas diffusion layer (GDL) porosity, CCL porosity, ionomer volume fraction, inlet temperature, initial membrane water content, catalyst layer (CL) surface area, and initial current, with respect to the uniformity of ice distribution and cold start failure time. Six of these parameters were chosen as inputs for a CNN-Transformers based neural network (CTnet), which was developed using Convolutional Neural Networks (CNN) and the multi-head attention mechanism of Transformers as a surrogate model for cold start. The optimization algorithm PSO-GA was used to find the optimal operating and geometric parameters, aiming to achieve the most uniform ice distribution in the CCL and extend the final cold start failure time. The optimized ice distribution uniformity was found to be 0.204446, showing a 1.1163 % improvement compared to the baseline model. The final cold start failure time was extended by 15.625 % to 185 s compared to the baseline model. The optimized model exhibited a more uniform distribution of ice during cold start, resulting in an extended cold start failure time. This indicates a reduced performance impact on the CCL, thereby improving the durability of the fuel cell. The optimization provides additional time for future auxiliary cold starts, while also enhancing the starting speed of auxiliary cold start through improved ice distribution uniformity.

Sentiment Analysis on Cyanide Case After 'Ice Cold' Aired with NLP Method using Naïve Bayes Algorithm

Information technology is developing increasingly rapidly, and the reach of the Internet has expanded even to remote areas. The public increasingly uses social media as a source of information that discusses all aspects of people's lives. Social media has a vital role for most people, one of which is the news of the cyanide coffee case. The Cyanide Coffee case was discussed again by netizens after Netflix raised this case in a documentary film entitled Ice Cold, which made the public even more convinced of the irregularities of the case. Based on this, sentiment analysis is needed to extract comments to obtain public opinion information. The sentiment analysis aims to create a sentiment model to determine public comments on this case. Therefore, this research was conducted to find out and classify public sentiment on the Cyanide Coffee Case using the Natural Language Processing (NLP) method, which is a text preprocessing process followed by the tokenization stage. Data filtering was used using Indonesian Stopwords, and then normalization was continued using Porter Stemmer. In this study, data collection was carried out based on public comments on Ice Cold shows on the TikTok platform using TikTok Comments Scraper. The test results show that the classification using naïve Bayes obtained the results of 22 negative comments, 4052 neutral comments and 34 positive comments. The classification results of this study are 87% accuracy, 97.6% precision, 87% recall, and 91.9% F-Score.

Far-Reaching Effects of Okhotsk Sea Ice Area on Sea Surface Heat Flux, Lower Atmosphere, and Ocean Mixed Layer

Abstract The impact of interannual variations in sea ice area in the Okhotsk Sea was investigated through a composite analysis of years with extensive and limited sea ice areas (referred to as heavy and light ice years, respectively), using atmospheric and oceanic reanalysis data. The comparison of heavy and light ice-year composites in February revealed a substantial decrease in upward surface turbulent heat flux in the Okhotsk Sea (∼−250 W m−2) and a notable increase in a surprisingly extensive region in the western North Pacific (30–120 W m−2), spanning 2300 km from the ice edge. These differences were consistent with the decrease in surface air temperature and specific humidity, suggesting that during heavy ice years, cold and dry air blowing from Siberia to the North Pacific via the Okhotsk Sea undergoes less modification over larger sea ice areas, remaining colder and drier in the North Pacific and thereby enhancing the heat flux. Such advection can be associated with the Asian winter monsoon and migratory cyclones. Cloud cover and surface radiation flux altered consistently with these differences, although longwave and shortwave radiation largely counterbalanced each other. Additionally, the Pacific storm track exhibited variation. In accordance with the heat flux difference, sea surface temperature decreased, and the ocean mixed layer deepened around the subarctic during heavy ice years. These findings suggest that sea ice area in the Okhotsk Sea influences the lower atmosphere and surface ocean in the North Pacific. Such impacts could further affect ocean nutrient circulation, ecosystems, and atmospheric teleconnections. Significance Statement Sea ice cover influences heat exchange between the atmosphere and ocean. Advection of less heated air over larger sea ice areas can enhance heat exchange downwind, and vice versa. We found that this was effective in the Okhotsk Sea, situated along the path of the Asian winter monsoon. Upward surface heat flux increased significantly across a vast region of the North Pacific associated with colder and drier air during years with larger sea ice areas. Additionally, cloud cover, Pacific storm track, sea surface temperature, and ocean mixed layer depth exhibited significant differences associated with sea ice area. These variations could impact ocean heat content, nutrient circulation, and primary production in the North Pacific, and atmospheric circulation in North America and Europe.

Jessica Wongso's Cyanide Coffee Case

The death of Wayan Mirna Salihin in 2016 shook Indonesia in its time. The Netflix documentary entitled "Ice Cold" has again brought Indonesian citizens to start talking again about this seven year old case. Indeed, the documentary is more inclined towards the coffee that Mirna drank not being proven to contain chemicals (cyanide). Which means the defendant, Jessica Wongso, cannot be completely blamed. This article includes analysis starting from the background, research methods, to the results and discussion. It may be noted that this article draws quite a bit of information from the Netflix documentary. Others were obtained from various official legal and scientific sources. This discussion will refer to the evidence collected, according to the methodology used. It will be discussed here that the Jessica Wongso Cyanide Coffee Case, even though it has passed, is still worthy of discussion in this nation.

X Netizens' Response to the Wayan Mirna Salihin Case after the Ice Cold Documentary Film was Released in George Gerbner's Model of Mass Communication

This research tries to analyze netizen Mirna Salihin's case's viral response in the mass media in 2016, and the whole public was talking about it. The case became popular again after the murder documentary was released on Netflix and sparked various discussions on social media. Netizens tried to find out what was considered odd about the murder. This research uses a qualitative descriptive method with netnographic analysis to explore social media phenomena. In this research, individuals or accounts on social media have the same capabilities as mass media. Individuals can summarize an event and choose which message (M1) will be shared with those closest to them or the public. The event summary is then spread to other individuals, triggering other opinions and being able to use the message (M1) to create other messages (M2), and so on. This research shows that the X Tirto.id account is a Man or Machine (M) who sees events and makes selections, then presents his views via social media, resulting in responses from other users such as the Bro Win and Ridwansyah Putra (E3) accounts as individuals who reinterpreted the Mirna Salihin case until other users responded again.

Film "Ice Cold: Murder, Coffee, and Jessica Wongso" Between Commercialization or New Direction

Film Ice Cold : Murder, Coffee, and Jessica Wongso akhir-akhir ini menjadi isu yang kontroversial di tengah masyrakat. Satu sisi menilai terdapat kejanggalan yang luar biasa yang harus di ungkap dalam kasus pembunuhan Wayang Mirna Salihin dengan kopi sianida hingga fil ini daat dijadikan petunjuk. Pada sisi lain, sebagaian masyrakat menilai film ini berpihak, menggiring opini publik, dan sebagai alat komersialisasi oleh pihak Netfflix. Penelitian ini bertujuan untuk menganalisis dua perspektif yang kontroversial di atas. Penelitian merupakan penelitian kualitiatf dengan pendekata deskriptif. Data yang digunakan dalam penelitian ini merupakan data sekunder yang didapatkan dari kutipan fil, sinopsis, buku, jurnal ilmiah, dan website yang kredibel. Data-data yang diukumpulkan dinalaissi dengan tahapan pengumpulan data, seleksi data, reduksi data, dan penarikan kesimpulan. Hasil penelitian menujukkan 1.) The film Ice Cold: Murder, Coffee, and Jessica Wongso can indirectly strengthen Jessica Wongso's side in filing for extraordinary legal action/judicial review which will then be constructed by constructing an argument as if the application for reconsideration this time is based on public unrest and complaints. 2.) The film Ice Cold: Murder, Coffee, and Jessica Wongso has the potential to become a commercialization tool because the film is paid, viral, and has cheap production costs compared to action films, horror films, and so on. 3.) Researchers agree with singer and actor Brissia Jodie that Netflix should make a part two film with a more balanced storyline on both sides.

A Paradigm Shift in Heart Preservation: Improved Post-transplant Outcomes in Recipients of Donor Hearts Preserved with the SherpaPak System.

Traditional ice storage has been the historic standard for preserving donor's hearts. However, this approach provides variability in cooling, increasing risks of freezing injury. To date, no preservation technology has been reported to improve survival after transplantation. The Paragonix SherpaPak Cardiac Transport System (SCTS) is a controlled hypothermic technology clinically used since 2018. Real-world evidence on clinical benefits of SCTS compared to conventional ice cold storage (ICS) was evaluated. Between October 2015 and January 2022, 569 US adults receiving donor hearts preserved and transported either in SCTS (n = 255) or ICS (n = 314) were analyzed from the Global Utilization And Registry Database for Improved heArt preservatioN (GUARDIAN-Heart) registry. Propensity matching and a subgroup analysis of >240 minutes ischemic time were performed to evaluate comparative outcomes. Overall, the SCTS cohort had significantly lower rates of severe primary graft dysfunction (PGD) (p = 0.03). When propensity matched, SCTS had improving 1-year survival (p = 0.10), significantly lower rates of severe PGD (p = 0.011), and lower overall post-transplant MCS utilization (p = 0.098). For patients with ischemic times >4 hours, the SCTS cohort had reduced post-transplant MCS utilization (p = 0.01), reduced incidence of severe PGD (p = 0.005), and improved 30-day survival (p = 0.02). A multivariate analysis of independent risk factors revealed that compared to SCTS, use of ice results in a 3.4-fold greater chance of severe PGD (p = 0.014). Utilization of SCTS is associated with a trend toward increased post-transplant survival and significantly lower severe PGD and MCS utilization. These findings fundamentally challenge the decades-long status quo of transporting donor hearts using ice.

Water Cooling, Ice Pack Application, and Cold Wet Compress Treatment of Burn Wounds in Children before Admission, Effects on Tissue Healing and Safety Research: Systematic Review and Meta-Analysis

Background: Burn injuries are caused by electricity, heat, radiation, cold, friction, or chemicals and lead to tissue destruction due to energy transfer from the sources to the tissues and cells. The present study aimed to perform a systematic review and meta-analysis of water cooling, ice pack application, cold and wet compress treatment of burn wounds in children before admission and to evaluate their effects on tissue healing and safety. Methods: We systematically searched PubMed, CINAHL, Cochrane Library, and Embase from inception to March 2023. Review Manager (version 5.4) was used to assess the risk of bias in the selected studies, and a meta-analysis of all dichotomous and continuous outcomes in the selected studies was performed. Out of 590 studies, seven studies based on the PRISMA protocol in the meta-analysis were included. Results: Based on the duration of cooling, no significant differences in the depth and size of burn wounds were found. Moreover, we established that cooling burn wounds significantly reduces tissue damage and limits the spread of burns to the surrounding tissues. A higher heterogeneity was observed in the selected studies based on methodology, implying different designs affecting our findings. Conclusion: There is inconclusive evidence on the recommended optimum duration of cooling burn wounds. However, cooling burn wounds has a beneficial impact on reducing tissue damage and limiting the spread of burns.

CARC21: Outcomes in Transplant Recipients Bridged with Impella Support: A GUARDIAN-Heart Subgroup Analysis

Background: The 2018 UNOS allocation changes prioritized recipients bridged with temporary MCS, which has recently increased in use. Also in 2018, the SherpaPak Cardiac Transport System (CTS) was introduced, which maintains donor heart temperatures continuously between 4°C - 8°C, potentially minimizing freezing-related myocardial injury. A subgroup analysis of the GUARDIAN Heart Registry examined outcomes in transplant recipients bridged using Impella. Methods: The GUARDIAN registry is an international, multicenter registry assessing outcomes after heart transplant comparing CTS preservation and traditional ice-cold (ICE) donor heart storage. Retrospective review of clinical outcomes in recipients bridged with an Impella were examined using summary statistics. Further analysis with ongoing enrollment will be available for presentation. Results: A total of 47 recipients with an Impella device received donor hearts preserved using either CTS (n=33) vs ICE (n=14). The two groups had similar baseline characteristics except the CTS cohort had significantly longer average ischemic time (175.3 vs and 223.9 minutes, p value = 0.02, ICE vs CTS,) and greater procurement distance travelled (170.2 vs 447.5 miles, p value < 0.001, ICE vs CTS). Post-transplant rates of severe PGD were low (14.3% vs 3.0%, p value = 0.15, ICE vs CTS, respectively). Survival was excellent with 100% 30-day survival in both cohorts. Conclusions: Despite significantly longer ischemic times by 48.6 minutes and over 2.5x farther organ procurement distance travelled, the use of the CTS was associated with excellent post-transplant outcomes in patients bridged using an Impella device. Considering ischemic time is a known risk factor for post-transplant PGD, these data support continued study of CTS use to optimize outcomes, in particular in an era of increasing distance travelled for organ procurement and complex bridging strategies.

Production of Two Novel Polyketide Pigments by Fusarium chlamydosporum

Aim: The aim of the present study was to evaluate the pigment production by the fungus Fusarium chlamydosporum in Glucose Peptone Yeast Extract medium and further to assess its toxicity by in vitro methods. Materials and Methods: The coloured crude extract yielded 4 fractions when subjected to column chromatography, upon which Fraction 2 and 4 were subjected to characterization procedures by UV-Visible spectrophotometry, Fourier Transform Infrared Spectroscopy and Molecular weight determination by Liquid Chromatography Mass Spectroscopy (LCMS). The toxicity of the pigments was evaluated in vitro on L-929 Mouse Fibroblast cell lines by MTT Assay. Results: The fungus produced dark red pigment in Glucose Peptone Yeast Extract medium on extraction with ice cold chloroform. The results summarized Fraction 2 and 4 to be the polyketide derivatives, Rubrofusarin and 8-O-methyl fusarubin respectively wherein the IC50 values of rubrofusarin and 8-O-methyl fusarubin was 183.85 and 82.366 μg/ml respectively. Conclusion: The significance of the present study lies in the characterization of 2 novel polyketides, rubrofusarin and 8-O-methyl fusarubin from F. chlamydosporum and there have been no literature reviews on the production of these pigments from the fungus.

A O03 Ex vivo perfusion of isolated human liver segments: the development of a novel model for ethical, translational research

Abstract Background Ex vivo perfusion techniques for human organs, in particular for the human liver, have been extensively studied for decades. Although ex vivo perfusion of human organs has been widely studied in the context of organ preservation and transplantation, it has also proven to be an invaluable tool in the development of novel models for translational pre-clinical research. Not only do these models allow more accurate study of human organ response to noxious external stimuli, but they also represent a far more ethical alternative to live animal experimentation.Although split-liver perfusion of the left or right hepatic lobe is well described in literature, ex vivo perfusion of isolated liver segments has not been previously attempted. A successful hepatic segmental perfusion model would provide a unique opportunity to study inflammation, response to infection and novel therapeutic approaches.The overall aim of this study was to establish an experimental ex vivo hepatic perfusion model on surgically resected human liver segments as a platform to evaluate and study organ preservation and function. The development of an ex vivo perfusion model of human liver segments would produce the ideal platform to study treatment effects without needing to sacrifice animals. Methods Patients were recruited as part of the TIMOLD (Tissue Models for Liver Disease) clinical trial (ClinicalTrials.gov Identifier: NCT05255042; 8/9/2021). Patients recruited were those aged eighteen or over undergoing elective liver resections at a single hepatobiliary unit. Nine human liver segments were retrieved following hemi-hepatectomy for colorectal liver metastases or hepatocellular carcinoma. A healthy segment was resected from the diseased specimen in theatre and a segmental hepatic artery and portal vein was cannulated immediately and flushed with ice cold heparinised preservation solution. Four segments were perfused with 600ml O negative expired red blood cells (HL-RBC) and five segments were perfused with 250ml Oxyglobin diluted with 250ml Volplex (HL-OXY). All segments were administered several drugs to provide metabolic support and optimise perfusion owing to their anticoagulant, antioxidant, and anti-inflammatory properties. Segments were perfused on a bespoke normothermic machine perfusion circuit for 4–6 hours. Perfusion parameters were monitored throughout perfusion and hourly hepatic venous blood gases were taken to monitor glucose and lactate levels. Hourly core biopsies were also taken for H&amp;E staining and interpreted by a consultant histopathologist. Data is reported as the median (range) and statistical analysis has been performed using the Mann-Whitney U test on Graphpad Prism. Results HL-RBC (n=4) had a median pre-perfusion weight of 216 (146–1330) grams whilst HL-OXY (n=5) had a median weight of 269(128–367) grams (p&amp;gt;0.05). HL-RBC segments had a median warm ischaemic (WI) time of 76.5(68–81) minutes and a median cold ischaemic (CI) time of 97 (35–119) minutes. HL-OXY segments had a median WI time of 5 (3–80) minutes and a median CI time of 38 (28–63) minutes (p&amp;gt;0.05). Median portal venous resistance was lower for the HL-OXY group (0.076mmHg/ml/min) in comparison to the HL-RBC group (0.62mmHg/ml/min); p&amp;lt;0.05. There was no significant difference in hepatic arterial resistance between the two groups (HL-RBC 3 mmHg/ml/min; HL-OXY 2.38 mmHg/ml/min; p&amp;gt;0.05). HL-OXY had a lower median lactate of 4.54mmol/L in comparison to HL-RBC (14.7mmol/L) throughout perfusion (p&amp;lt;0.05). HL-OXY segments also had a lower median glucose of 5.9mmol/L compared to 9.65mmol/L for HL-RBC (p&amp;lt;0.05). 1/4 HL-RBC segments had normal histology after 6 hours of perfusion. 2/4 segments showed evidence of necrosis after 2 hours (one with established cirrhosis), and 1/4 segment demonstrated necrosis after 3 hours of perfusion. 4/5 HL-OXY segments demonstrated normal histology after 6 hours of perfusion. One segment, with known cirrhosis, demonstrated necrosis after two hours of perfusion. Conclusions We present here, for the first time, the development of a successful ex vivo isolated hepatic segmental perfusion model. We demonstrate, in the development of this model, that bovine haemoglobin glutamer-200 (Oxyglobin) is a superior oxygen carrier to expired O negative red blood cells in the ex vivo perfusion of isolated human hepatic segments. The HL-OXY group demonstrated a significantly lower portal venous resistance in comparison to HL-RBC, although there was no difference in arterial resistance. Furthermore, HL-OXY segments had lower median glucose and lactate levels during perfusion compared to HL-RBC segments. 4 of 5 segments perfused with Oxyglobin demonstrated viable hepatocytes on histopathological analysis after 6 hours of perfusion. The segment demonstrating early necrosis was one with established cirrhosis on pre-perfusion histology. We describe a protocol for the successful ex vivo perfusion of non-cirrhotic human hepatic segments. Human liver is routinely resected in tertiary hepatobiliary units and much of the specimen discarded. The paradigm presented here identifies a readily available ethical source of human liver with minimal warm ischaemia. This facilitates optimal conditions for translational research, is cost-effective and avoids animal experimentation.