Postmortem Liver Research Articles

A Stability Study of Atropine in Post-Mortem Peripheral Blood and Liver at -20°C, +4°C, +10°C and +20°C Over 12 Months. Peripheral Blood and Liver Concentration Ratios

At the judicial inquiry of a criminal case involving atropine, we were required to carry out toxicological analyses on a body exhumed 31 months after death. It was obviously not possible to take samples of peripheral blood or any other fluid at the autopsy of the victim.

Disseminated pleomorphic rhabdomyosarcoma in a horse.

An 8-y-old National Show Horse mare was presented for evaluation of pneumonia and laminitis. Harsh bronchovesicular sounds were auscultated throughout both lung fields, and the mare had signs of moderately painful laminitis. Thoracic ultrasonography revealed lung consolidation throughout the dorsal aspect of both lungs, and radiography revealed an extensive diffuse-to-patchy bronchointerstitial lung pattern. The mare's clinical condition rapidly deteriorated, and euthanasia was elected. On postmortem examination, the lungs, omentum, spleen, liver, adrenal glands, kidneys, and femur contained 0.5-2.5-cm, firm, tan nodules. Histologically, the lungs, spleen, liver, kidneys, adrenal glands, omentum, left eye, and femur were infiltrated by bundles and nests of pleomorphic polygonal-to-spindloid cells intermixed with frequent multinucleate cells. Lymphatic vessels in the affected tissues were frequently distended with tumor emboli. Neoplastic cells were diffusely positive for vimentin, desmin, sarcomeric actin, myoblastic differentiation protein 1, and myogenin, supportive of the diagnosis of rhabdomyosarcoma (RMS), which is a rare neoplasm in horses. Cross-striations were not evident with H&E or phosphotungstic acid-hematoxylin stains. Markedly pleomorphic neoplastic cells, multinucleate cells, and lack of cross-striations suggested the subclassification of pleomorphic RMS.

Tissue specific responses to oxidative fuel source preference during heat stress in lactating dairy cows

Prolonged exposure to high environmental temperatures results in an accumulated heat load that induces a heat stress (HS) response in dairy cattle. Heat stress compromises dairy farm profitability by reducing milk yield, altering milk composition, and hindering reproductive performance. The ability to alternate between carbohydrate and lipid sources for energy production is termed metabolic flexibility (Met Flex). The objective of this study was to evaluate the Met Flex of mammary, muscle, and liver tissue in lactating dairy cows under HS and thermoneutral (TN) conditions. Sixteen Holstein cows were assigned to 1 of 2 treatment groups: pair feeding in a TN environment (PFTN) or HS. All cows experienced a 4 d TN period with ad libitum intake followed by a 4 d treatment period. Heat stress cows were exposed to a temperature humidity index (THI) ranging from 76 to 80 and the PFTN cows were exposed to a THI of 64. Milk production and health data were recorded twice daily. Semitendinosus biopsies were obtained on d 4 of each period and post-mortem mammary and liver samples were obtained on d 4 of period 2. All tissue samples were assayed for Met Flex. Activity of mitochondrial (Mit) enzymes were assessed in skeletal muscle only. Four d of HS decreased milk yield, altered milk composition, and increased respiration rate and rectal temperatures. No differences in Met Flex were observed in mammary or liver tissue during period 2. However, HS, but not PFTN conditions, lowered Met Flex of skeletal muscle by 18.3% when compared with TN ad libitum feed intake conditions of period 1. No treatment differences were observed in skeletal muscle Mit enzyme activity indicating the decrease in Met Flex occurred independently of changes in Mit function. The reduction in Met Flex of skeletal muscle during HS may contribute to reduced milk yield and warrants further investigation.

Postmortem liver hypostases in newborns: radiation and pathological characteristics

Postmortem liver hypostases in newborns: radiation and pathological characteristics

Natural history study of hepatic glycogen storage disease type IV and comparison to Gbe1ys/ys model.

BackgroundGlycogen storage disease type IV (GSD IV) is an ultrarare autosomal recessive disorder that causes deficiency of functional glycogen branching enzyme and formation of abnormally structured glycogen termed polyglucosan. GSD IV has traditionally been categorized based on primary hepatic or neuromuscular involvement, with hepatic GSD IV subclassified as discrete subtypes: classic (progressive) and nonprogressive.MethodsTo better understand the progression of liver disease in GSD IV, we present clinical and histopathology data from 23 patients from around the world and characterized the liver involvement in the Gbe1ys/ys knockin mouse model.ResultsWe propose an alternative to the established subtype-based terminology for characterizing liver disease in GSD IV and recognize 3 tiers of disease severity: (i) "severe progressive" liver disease, (ii) "intermediate progressive" liver disease, and (iii) "attenuated" liver disease. Analysis of liver pathology revealed that risk for liver failure cannot be predicted from liver biopsy findings alone in individuals affected by GSD IV. Moreover, analysis of postmortem liver pathology from an individual who died over 40 years after being diagnosed with nonprogressive hepatic GSD IV in childhood verified that liver fibrosis did not regress. Last, characterization of the liver involvement in a mouse model known to recapitulate the adult-onset neurodegenerative form of GSD IV (Gbe1ys/ys mouse model) demonstrated hepatic disease.ConclusionOur findings challenge the established subtype-based view of GSD IV and suggest that liver disease severity among patients with GSD IV represents a disease continuum.Trial registrationClinicalTrials.gov NCT02683512FundingNone.

Limits and pitfalls in a lethal infant cocaine intoxication

ABSTRACT The case concerns the first Italian report of a fatal cocaine intoxication in an 8-month-old baby girl found dead in the cradle by her parents at 6 am on a summer day. At arrival to the emergency department, the doctors declared the baby dead and the Prosecutor Office ordered an autopsy and toxicological analyses. Despite no history of cocaine use/abuse by both parents, toxicological analyses evidenced a positivity to cocaine in the post-mortem blood, brain and liver. The legal physician concluded for a fatal cocaine intoxication. A crucial aspect to the Court was to discriminate between administration (accidental or voluntary) and intake (almost accidentally), due to different legal consequences for parents (i.e. voluntary homicide or inadequate supervision). In the here presented case discriminating the exact intake was almost impossible in terms of certainty. Such difficulty mainly derived from lacks in the investigation activities in the first hours following the death. Among all possible intake routes, the ‘most probable’ was the accidental one, mostly attributable to a cradle contamination. A lack in the site inspection protocol could strongly affect the case solution making it impossible to offer the Court a definitive ‘scientific’ solution to the case (based on scientifically irrefutable data).

Searching for homozygous haplotype deficiency in Manech Tête Rousse dairy sheep revealed a nonsense variant in the MMUT gene affecting newborn lamb viability

BackgroundRecessive deleterious variants are known to segregate in livestock populations, as in humans, and some may be lethal in the homozygous state.ResultsWe used phased 50 k single nucleotide polymorphism (SNP) genotypes and pedigree data to scan the genome of 6845 Manech Tête Rousse dairy sheep to search for deficiency in homozygous haplotypes (DHH). Five Manech Tête Rousse deficient homozygous haplotypes (MTRDHH1 to 5) were identified, with a homozygous deficiency ranging from 84 to 100%. These haplotypes are located on Ovis aries chromosome (OAR)1 (MTRDHH2 and 3), OAR10 (MTRDHH4), OAR13 (MTRDHH5), and OAR20 (MTRDHH1), and have carrier frequencies ranging from 7.8 to 16.6%. When comparing at-risk matings between DHH carriers to safe matings between non-carriers, two DHH (MTRDHH1 and 2) were linked with decreased insemination success and/or increased stillbirth incidence. We investigated the MTRDHH1 haplotype, which substantially increased stillbirth rate, and identified a single nucleotide variant (SNV) inducing a premature stop codon (p.Gln409*) in the methylmalonyl-CoA mutase (MMUT) gene by using a whole-genome sequencing approach. We generated homozygous lambs for the MMUT mutation by at-risk mating between heterozygous carriers, and most of them died within the first 24 h after birth without any obvious clinical symptoms. Reverse transcriptase-qPCR and western blotting on post-mortem liver and kidney biological samples showed a decreased expression of MMUT mRNA in the liver and absence of a full-length MMUT protein in the mutant homozygous lambs.ConclusionsWe identified five homozygous deficient haplotypes that are likely to harbor five independent deleterious recessive variants in sheep. One of these was detected in the MMUT gene, which is associated with lamb lethality in the homozygous state. A specific management of these haplotypes/variants in the MTR dairy sheep selection program would help enhance the overall fertility and lamb survival.

A primary study of ethanol production in postmortem liver and muscle tissue of rats

ObjectiveTo study the characteristics of postmortem ethanol production and its relation with alcohol congeners in postmortem rat liver and muscle tissues. MethodPostmortem liver and muscle tissues in Sprague–Dawley rats, from postmortem time interval (PMI) day 0–20, were analyzed via headspace gas chromatograph flame ionization detection to observe production of postmortem ethanol and 5 selected alcohol congeners. Result1. Putrid ethanol production increased gradually to a peak and then decreased with the prolongation of PMI; 2. Acetaldehyde, 1-propanol, and 3-methyl-butyraldehyde were produced along with postmortem ethanol; 1-butanol was only detected from day 11–20; 3. The concentrations of acetaldehyde, 1-propanol and 3-methyl-butyraldehyde was related with ethanol production. Fifteen mathematical models were constructed for putrid ethanol production based on acetaldehyde, 1-propanol, and 3-methyl-butyraldehyde. ConclusionA peak in postmortem ethanol production was identified. The production trends of acetaldehyde, 1-propanol, and 3-methyl-butyraldehyde in the liver, and of 1-propanol in muscle, were consistent with those of ethanol, and could potentially to be used as biomarkers of postmortem ethanol production. Further human samples and data analysis are needed to verify this.

Ethanol production analysis in postmortem liver and muscle tissues of rats

Ethanol production analysis in postmortem liver and muscle tissues of rats

Correlation between interleukin-6 expression in post-mortem core liver biopsy and degree of liver injury in patients with fatal COVID-19.

Excessive release of interleukin-6 (IL-6) during the progression of coronavirus disease 2019 (COVID-19) induces cytokine storms, resulting in multi-organ damages including liver injury, similar in nature with mechanism of viral hepatitis. Systemic IL-6 has been associated with the incidence of liver injury among COVID-19 patients; however, studies on IL-6 expression in the liver tissue are completely lacking. The aim of this study was to measure the IL-6 expression in the liver tissues and to determine its correlation with the degree of liver injury in fatal COVID-19 patients. Through this first cross-sectional study, IL-6 expression was measured through immunohistochemical staining and the degree of liver injury was identified based on level of serum alanine aminotransferase (ALT). The Spearman correlation test was used to identify the correlation between IL-6 expression and the degree of liver injury. A total of 47 deceased COVID-19 patients were included and IL-6 expression was observed in all post-mortem liver specimens, ranging from mild to strong expression. Liver injury at various degrees (mild to severe) was found in more than half (59.5%) of the cases. The Spearman correlation analysis suggested a statistically insignificant correlation between liver IL-6 expression and the degree of liver injury (r=0.152; p=0.309). In conclusion, even IL-6 expression was observed in all post-mortem liver specimens, there was an insignificant correlation between IL-6 expression in the liver tissue with the degree of liver injury among fatal COVID-19 patients, suggesting that IL-6 was not the only main factor contributing to liver damage in COVID-19 patients.

Immunoassay testing forbarbiturates using alternative matrices inpostmortem tissues from cats anddogs.

The barbiturate drug pentobarbital is commonly used by veterinarians for the euthanasia of domestic animals. During the veterinary forensic autopsy, it is sometimes necessary to determine whether the animal was chemically euthanized with pentobarbital. The use of a human immunochromatographic test for barbiturate screening utilizing dog or cat urine has been previously validated; however, the use of alternative matrices for this purpose is yet to be explored when urine is not available. Postmortem heart, liver, spleen, skeletal muscle, blood and/or urine samples from 20 dogs and 26 cats with a reported chemical euthanasia status were processed using two different methods, bead homogenization and sonication, and screened for barbiturates using a human immunochromatographic test. There was 100% agreement of the immunochromatographic test results using the sonication method with the reported euthanasia status of both dogs and cats. Using the bead homogenization method, agreement with the reported euthanasia status was 93.3% and 96.7% for dogs and cats, respectively, due to invalid test results from four dog and two cat samples. A subset of liver samples (10 canine and 10 feline) was analyzed via gas chromatography-mass spectrometry, and there was 100% agreement between the immunochromatographic test results and gas chromatography-mass spectrometry results for both cats and dogs. Overall, our results support the use of a variety of alternative matrices for barbiturate screening in cats and dogs.

PSXIV-7 Comparison of Detection of Fasciola in Naturally Infected Slaughter (dairy) Cattle By Postmortem Examination, Sedimentation, Antibody Elisa and Real-Time Pcr

Abstract Fasciolosis is a global food-borne disease affecting livestock. Management of the parasitic disease greatly depends on control and proper monitoring through early detection. Highly improved diagnostic methods to detect fasciolosis have been introduced. Limited research has paid attention to comparing four detection methods. The current study aimed to compare the detection of Fasciola in naturally infected slaughtered dairy cattle using four methods. Blood and fecal samples were collected from slaughtered cattle in three high throughput abattoirs in the Eastern Cape Province, South Africa, from 2020 through 2021. Postmortem liver examination was conducted to identify Fasciola spp. Then blood from condemned carcasses were analyzed using antibody ELISA to detect Fasciola antibodies, sedimentation technique to detect Fasciola eggs; and quantitative real-time PCR (qPCR) to detect Fasciola DNA. A binomial Generalized Linear Model (GLM) and Cohen’s Kappa test were used to compare the detection rate and level of agreement across the four diagnostic methods. Significant differences (P = 0.0001) across the four diagnostic methods existed. Postmortem liver examination (94.5%) and qPCR (90.4%) compared ELISA (22.6%) and sedimentation (3.53%), respectively. There was a poor agreement between sedimentation and Ab-ELISA (kappa= -0.105, P = 0.128), sedimentation and postmortem liver examination (kappa= -0.054, P = 0.163); and qPCR and liver examination (kappa= -0.076, P = 0.341) In conclusion, results from the current study revealed significant differences among the four detection methods. The postmortem liver examination and qPCR were more reliable, sensitive, and accurate than ELISA and sedimentation.

Infectious complications as prognostic factors for negative outcomes of liver transplantation

Aim. To study the influence of infectious complications on the outcomes of orthotopic liver transplantation.Materials and methods. The methodology involved analyzing the outcomes of 159 orthotopic transplantations of full postmortem liver for 2013–2022. Surgical complications were classified according to Clavien-Dindo (2004). Special attention was paid to microbiological examination of biological fluids with determination of the pathogen species and susceptibility to antimicrobial agents. Determination of isolated cultures and sensitivity to antibacterial agents was carried out using automatic analyzers. Sensitivity and resistance to antimicrobials were determined according to the recommendations of the European Committee (EUCAST, www.eucast.org).Results. At present, 78 of 141 recipients are alive. After discharge, 42 died of various causes within 4–124 months. In-hospital mortality was 9.9% after primary liver transplantations and 38.9% after retransplantations. Recently, K. pneumoniae (18.8%), E. coli (18.7%), C. albicans (9.7%) have dominated the pathogen spectrum. In microbiological examination, after primary transplantation, pathogens were more often isolated from the wound discharge, after retransplantation – from the biliary system.Conclusion. Infectious complications remain a significant cause of unsatisfactory outcomes of liver transplantation. The greatest negative prognostic role belongs to biliary infection caused by multidrug-resistant pathogens in combination with insufficient arterial blood supply of the transplant.

Fatalities Involving Khat in Jazan, Saudi Arabia, 2018 to 2021.

Interpreting fatalities involving khat is challenging due to a lack of data on cathinone and cathine reference concentrations in postmortem tissues. This study investigated the autopsy findings and toxicological results of fatalities involving khat in Saudi Arabia's Jazan region from 1 January 2018 to 31 December 2021. All confirmed cathine and cathinone results in postmortem blood, urine, brain, liver, kidney, and stomach samples were recorded and analyzed. Autopsy findings and the manner and cause of death of the deceased were assessed. Saudi Arabia's Forensic Medicine Center investigated 651 fatality cases over four years. Thirty postmortem samples were positive for khat's active constituents, cathinone and cathine. The percentage of fatalities involving khat was 3% in 2018 and 2019 and increased from 4% in 2020 to 9% in 2021, when compared with all fatal cases. They were all males ranging in age from 23 to 45. Firearm injuries (10 cases), hanging (7 cases), road traffic accident (2 cases), head injury (2 cases), stab wounds (2 cases), poisoning (2 cases), unknown (2 cases), ischemic heart disease (1 case), brain tumor (1 case), and choking (1 case) were responsible for the deaths. In total, 57% of the postmortem samples tested positive for khat only, while 43% tested positive for khat with other drugs. Amphetamine is the drug most frequently involved. The average cathinone and cathine concentrations were 85 and 486 ng/mL in the blood, 69 and 682 ng/mL in the brain, 64 and 635 ng/mL in the liver, and 43 and 758 ng/mL in the kidneys, respectively. The 10th-90th percentiles of blood concentrations of cathinone and cathine were 18-218 ng/mL and 222-843 ng/mL, respectively. These findings show that 90% of fatalities involving khat had cathinone concentrations greater than 18 ng/mL and cathine concentrations greater than 222 ng/mL. According to the cause of death, homicide was the most common fatality involving khat alone (77%). More research is required, especially toxicological and autopsy findings, to determine the involvement of khat in crimes and fatalities. This study may help forensic scientists and toxicologists investigate fatalities involving khat.

Steatosis and Steatohepatitis in Autopsy Specimens of Liver

Background: Liver is the site of many diseases, many of which become symptomatic while some are diagnosed only on autopsy. Involvement of liver is secondary to cardiac, metabolic, social problem like alcoholism, drug toxicity, herbal remedies and environmental exposures. Objectives: This study aims at finding the frequency of steatosis and steatohepatitis in autopsy specimens of liver. Methods: This was a descriptive cross-sectional study carried out at the Department of Pathology, Dhaka Medical College, Dhaka over a period of 2011-2012. Liver samples of postmortem cases from the Department of Forensic Medicine, Dhaka Medical College and autopsy samples of liver received at the Department of Pathology, Dhaka Medical College, sent from different parts of Bangladesh via postal service for histopathological examination were included in this study. Result: Satisfactory liver tissue samples for histological evaluation were available in 200 autopsy specimens (155 male and 45 female) with a mean age of 39.50±14.76 years. Out of 200 cases, the most common lesion was steatosis in 35(17.50%) cases. Steatosis mild, moderate and severe were found in 20(10%) cases, 9(4.50%) cases and 6(3.0%) cases respectively. Steatohepatitis was found in 16(8.0%) cases. Steatohepatitis with fibrous portal expansion, steatohepatitis with bridging fibrosis and steatohepatitis with cirrhosis were detected in 7(3.5%), 6(3%) and 3(1.5%) cases respectively. Conclusion: A high prevalence of steatosis and steatohepatitis was detected in postmortem liver samples of Bangladesh. Since both diseases can have serious clinical consequences, they should be considered as an important threat to the health of the general population of our country.

COVID-19-related hyperglycemia is associated with infection of hepatocytes and stimulation of gluconeogenesis

Occurrence of hyperglycemia upon infection is associated with worse clinical outcome in COVID-19 patients. However, it is still unknown whether SARS-CoV-2 directly triggers hyperglycemia. Herein, we interrogated whether and how SARS-CoV-2 causes hyperglycemia by infecting hepatocytes and increasing glucose production. We performed a retrospective cohort study including patients that were admitted at a hospital with suspicion of COVID-19. Clinical and laboratory data were collected from the chart records and daily blood glucose values were analyzed to test the hypothesis on whether COVID-19 was independently associated with hyperglycemia. Blood glucose was collected from a subgroup of nondiabetic patients to assess pancreatic hormones. Postmortem liver biopsies were collected to assess the presence of SARS-CoV-2 and its transporters in hepatocytes. In human hepatocytes, we studied the mechanistic bases of SARS-CoV-2 entrance and its gluconeogenic effect. SARS-CoV-2 infection was independently associated with hyperglycemia, regardless of diabetic history and beta cell function. We detected replicating viruses in human hepatocytes from postmortem liver biopsies and in primary hepatocytes. We found that SARS-CoV-2 variants infected human hepatocytesin vitrowith different susceptibility. SARS-CoV-2 infection in hepatocytes yields the release of new infectious viral particles, though not causing cell damage. We showed that infected hepatocytes increase glucose production and this is associated with induction of PEPCK activity. Furthermore, our results demonstrate that SARS-CoV-2 entry in hepatocytes occurs partially through ACE2- and GRP78-dependent mechanisms. SARS-CoV-2 infects and replicates in hepatocytes and exerts a PEPCK-dependent gluconeogenic effect in these cells that potentially is a key cause of hyperglycemia in infected patients.

“Spice”-related deaths in and around Munich, Germany: A retrospective look at the role of synthetic cannabinoid receptor agonists in our post-mortem cases over a seven-year period (2014–2020)

Synthetic cannabinoid receptor agonists (SCRAs, “Spice”) are a diverse group of recreational drugs, with their structural and pharmacological variability still evolving. Forensic toxicologists often rely on previous reports to assess their role in intoxication cases. This work provides detailed information on the “Spice”-related fatalities around Munich, Germany, from 2014 to 2020. All cases underwent an autopsy. Pharmaceutical and illicit drugs were detected and quantified in post-mortem peripheral blood or liver by liquid chromatography-tandem mass spectrometry (LC–MS/MS). Based on circumstantial evidence, only those cases for which a prior consumption was suspected underwent additional analyses for SCRAs and other new psychoactive substances in post-mortem blood, liver or antemortem specimens. Drug concentrations, pathological findings at autopsy and case histories were considered to assess and rank the SCRAs’ involvement in each death. Concentration ranges for the individual substances in blood were defined and their distribution patterns over the investigated period were determined and correlated with their legal status and local police seizures. We identified 41 different SCRAs among 98 fatalities. 91.8% were male, at a median age of 36 years. SCRAs played a causative role in 51%, contributory role in 26%, and an insignificant role in 23% of cases. In correlation with local police seizures and legal status, 5F-ADB was the most prevalent in our cases, followed by 5F-MDMB-PICA and AB-CHMINACA. Cumyl-CBMICA and 5F-MDMB-P7AICA were among the least frequently detected SCRAs. “Spice”-related fatalities and SCRAs’ causative role have significantly decreased among our cases since the German New Psychoactive Substances Act.

Liver Injury Associated with COVID-19 Infection: Pathogenesis, Histopathology, Prognosis, and Treatment

Liver injury occurs frequently as a consequence of SARS-CoV-2 infection. Direct infection of the liver leads to hepatic impairment with elevated transaminases. In addition, severe COVID-19 is characterized by cytokine release syndrome, which may initiate or exacerbate liver injury. In patients with cirrhosis, SARS-CoV-2 infection is associated with acute-on-chronic liver failure. The Middle East and North Africa (MENA) region is one of the world's regions characterized by a high prevalence of chronic liver diseases. Both parenchymal and vascular types of injury contribute to liver failure in COVID-19, with a myriad of pro-inflammatory cytokines playing a major role in perpetuating liver injury. Additionally, hypoxia and coagulopathy complicate such a condition. This review discusses the risk factors, and the underlying causes of impaired liver functions in COVID-19, with a focus on key players in the pathogenesis of liver injury. It also highlights the histopathological changes encountered in postmortem liver tissues as well as potential predictors and prognostic factors of such injury, in addition to the management strategies to ameliorate liver damage.

Evidence of SARS-CoV-2 infection in postmortem lung, kidney, and liver samples, revealing cellular targets involved in COVID-19 pathogenesis

There is an urgent need to understand severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-host interactions involved in virus spread and pathogenesis, which might contribute to the identification of new therapeutic targets. In this study, we investigated the presence of SARS-CoV-2 in postmortem lung, kidney, and liver samples of patients who died with coronavirus disease (COVID-19) and its relationship with host factors involved in virus spread and pathogenesis, using microscopy-based methods. The cases analyzed showed advanced stages of diffuse acute alveolar damage and fibrosis. We identified the SARS-CoV-2 nucleocapsid (NC) in a variety of cells, colocalizing with mitochondrial proteins, lipid droplets (LDs), and key host proteins that have been implicated in inflammation, tissue repair, and the SARS-CoV-2 life cycle (vimentin, NLRP3, fibronectin, LC3B, DDX3X, and PPARγ), pointing to vimentin and LDs as platforms involved not only in the viral life cycle but also in inflammation and pathogenesis. SARS-CoV-2 isolated from a patient´s nasal swab was grown in cell culture and used to infect hamsters. Target cells identified in human tissue samples included lung epithelial and endothelial cells; lipogenic fibroblast-like cells (FLCs) showing features of lipofibroblasts such as activated PPARγ signaling and LDs; lung FLCs expressing fibronectin and vimentin and macrophages, both with evidence of NLRP3- and IL1β-induced responses; regulatory cells expressing immune-checkpoint proteins involved in lung repair responses and contributing to inflammatory responses in the lung; CD34+ liver endothelial cells and hepatocytes expressing vimentin; renal interstitial cells; and the juxtaglomerular apparatus. This suggests that SARS-CoV-2 may directly interfere with critical lung, renal, and liver functions involved in COVID-19-pathogenesis.

Portable and green solid contact potentiometric sensor for the rapid and direct assay of clozapine in post-mortem rat liver and dosage forms: An analytical approach to forensic and pharmaceutical samples

This work is devoted to fabricating and validating a portable potentiometric sensor for in-site clozapine (CLZ) assay in tablets and post-mortem rat liver. The outcomes marking the overall sensor performance were defined to be (Nernstian slope, quantification limit, and correlation coefficients). A polyaniline-coated glassy carbon electrode was used as support for the sensors. The optimized sensor was composed of PVC (30.85 % w/w) plasticized with 2-nitrophenyl phenyl ether NPPE (64.32 % w/w) incorporating phosphotungstic acid PT (1.61 % w/w) as a cationic exchanger and calix-8-arene CX-8 (3.22 % w/w) as an ionophore, where the membrane thickness was 100 µm. The developed membrane works over the pH range of 2.3–6, providing a dynamic response time of 8 s. A computational docking approach was implemented, and the ionophore-CLZ complex interactions were studied to identify the best-fit complex. The docking study demonstrates that CX-8 is the optimum choice since it binds with CLZ+ by forming four bonds with −4.36 as binding free energy, reflecting the expected high stability of the developed complex. Validation of the proposed sensor illustrates that CLZ follows a linear response over the range of 1.8 × 10−5–1 × 10−2 M, yielding a correlation coefficient value of 0.9989 and a limit of detection LOD of 2.0 × 10−6 M. Moreover, the constructed membrane sensor proves to be of acceptable accuracy as reflected by the recovery percentage (97.48 %) and provides satisfactory precision as shown by the low relative standard deviation values; 0.114 for intermediate precision and 0.772 for repeatability. The greenness profile of the proposed method was assessed by the Green Analytical Procedure Index (GAPI), compared with the reported HPLC method, and found superior.