The effects of cryopreservation on human hepatocytes obtained from different sources of liver tissue

Successful cryopreservation of human hepatocytes is important to establish hepatocyte banks for clinical use or in vitro research. The availability of donor tissue from unused liver segments/lobes and non-heart-beating donors (NHBD) has provided newer sources of hepatocytes. The quality of hepatocytes at the time of cryopreservation is important as cells isolated from liver tissue of borderline quality may not withstand the stresses associated with cryopreservation and subsequent thawing. Human hepatocytes were cryopreserved after isolation from mainly donor tissues (n = 40). In vitro assessment of the viability and function of the fresh and thawed cryopreserved hepatocytes was performed. Viability, attachment efficiency, enzyme activity, and albumin production of hepatocytes were all significantly decreased, and LDH leakage significantly increased, on thawing after cryopreservation. The viability of cryopreserved hepatocytes isolated from tissue rejected for orthotopic liver transplantation (36 +/- 15%) was significantly lower than those isolated from tissue where part was used for liver transplantation (47 +/- 14%, p = 0.002), but there were no significant differences in functional parameters. The viability of cryopreserved hepatocytes isolated from NHBD tissue (29 +/- 9%, p = 0.001) and from steatotic donor tissue (35 +/- 11%, p = 0.019) was significantly lower than those isolated from normal donor tissue (49 +/- 14%). There was no difference in functional parameters, except for albumin production of hepatocytes from NHBD tissue (2.9 +/- 1.0 microg/h/mg protein) being significantly lower than those from normal donor tissue (4.8 +/- 2.8 microg/h/mg protein, p = 0.03). The viability and attachment efficiency of cryopreserved hepatocytes isolated from liver tissue from resections for tumors was significantly higher, and the LDH leakage significantly lower, than those isolated from all donor tissue. Hepatocytes isolated from NHBD and steatotic tissue were more vulnerable to the effects of cryopreservation. Further research is required to improve hepatocyte isolation and cryopreservation protocols for different types of liver tissue.     

Phylogenetic Analysis of Spread of Hepatitis C Virus Identified during HIV Outbreak Investigation,Unnao, India

An HIV outbreak investigation during 2017–2018 in Unnao District, Uttar Pradesh, India, unearthed high prevalence of hepatitis C virus (HCV) antibodies among the study participants. We investigated these HCV infections by analyzing NS5B and core regions. We observed no correlation between HIV–HCV viral loads and clustering of HCV sequences, regardless of HIV serostatus. All HCV isolates belonged to genotype 3a. Monophyletic clustering of isolates in NS5B phylogeny indicates emergence of the outbreak from a single isolate or its closely related descendants. The nucleotide substitution rate for NS5B was 6 × 10⁻³ and for core was 2 × 10⁻³ substitutions/site/year. Estimated time to most recent common ancestor of these isolates was 2012, aligning with the timeline of this outbreak, which might be attributable to unsafe injection practices while seeking healthcare. HIV–HCV co-infection underlines the need for integrated testing, surveillance, strengthening of healthcare systems, community empowerment, and molecular analyses as pragmatic public health tools.     

Filarial infection is resisted differentially by subjects having different blood group phenotypes

This study was aimed to elucidate the relationship between major blood group antigens (BGAs) and susceptibility or resistance to human lymphatic filariasis. A total of 492 human subjects, living in Raipur city of Chhattisgarh, endemic for bancroftian filariasis in Central India, were screened for the presence of Wuchereria bancrofti microfilariae and disease manifestations. The frequency of BGAs was tabulated as a function of the status of filariasis, namely normal (no infection), mf carrier (presence of microfilariae in blood), and elephantiasis (confirmed clinical manifestations). The comparison of ABO phenotype distributions among all the three groups clearly indicated that disease status has a significant relationship with the blood group attributes. The result of χ² analysis of the frequencies of ABO phenotypes observed in microfilaraemic and elephantiasis groups (observed frequency vs. expected frequency computed based on the distribution of the normal population) revealed that there has been a significant alteration in the distribution of ABO phenotypes in microfilaraemic, but not in elephantiasis, group. The susceptibility to filarial infection was computed as a function of blood group phenotypes in the population. The susceptibility and conditional susceptibility for elephantiasis were the least among the subjects with AB phenotypes. The mechanism of association between filariasis and ABO antigens needs to be further explored to understand how the distribution of BGAs affects susceptibility/resistance to infection. J. Clin. Lab. Anal. 23:186–191, 2009. © 2009 Wiley‐Liss, Inc.     

A new look at glutamate and ischemia: NMDA agonist improves long-term functional outcome in a rat model of stroke

Ischemic stroke triggers a massive, although transient, glutamate efflux and excessive activation of NMDA receptors (NMDARs), possibly leading to neuronal death. However, multiple clinical trials with NMDA antagonists failed to improve, or even worsened, stroke outcome. Recent findings of a persistent post-stroke decline in NMDAR density, which plays a pivotal role in plasticity and memory formation, suggest that NMDAR stimulation, rather than inhibition, may prove beneficial in the subacute period after stroke. AIM: This study aims to examine the effect of the NMDAR partial agonist d-cycloserine (DCS) on long-term structural, functional and behavioral outcomes in rats subjected to transient middle cerebral artery occlusion, an animal model of ischemic stroke. MATERIALS #ENTITYSTARTX00026; METHODS: Rats (n = 36) that were subjected to 90 min of middle cerebral artery occlusion were given a single injection of DCS (10 mg/kg) or vehicle (phosphate-buffered saline) 24 h after occlusion and followed up for 30 days. MRI (structural and functional) was used to measure infarction, atrophy and cortical activation due to electrical forepaw stimulation. Memory function was assessed on days 7, 21 and 30 postocclusion using the novel object recognition test. A total of 20 nonischemic controls were included for comparison. RESULTS: DCS treatment resulted in significant improvement of somatosensory and cognitive function relative to vehicle treatment. By day 30, cognitive performance of the DCS-treated animals was indistinguishable from nonischemic controls, while vehicle-treated animals demonstrated a stable memory deficit. DCS had no significant effect on infarction or atrophy. CONCLUSION: These results support a beneficial role for NMDAR stimulation during the recovery period after stroke, most likely due to enhanced neuroplasticity rather than neuroprotection.     

Neonatal Purpura Fulminans Due to Methicillin Resistant Staphylococcus aureus

Abstract: Neonatal purpura fulminans is rare and may be inherited or acquired. It may ultimately lead to multiorgan failure and death. Purpura fulminans in a premature neonate resulting from Staphylococcus aureus septicemia is illustrated. Unfortunately, the baby succumbed to septicemia.     

Iodine deficiency disorders in school age children in Kullu district, Himachal Pradesh

Iodine deficiency disorder is a major public health problem in Himachal Pradesh. A study was conducted in district Kullu to assess the prevalence of IDD in school age children. Clinical examination of the thyroid of 1986 children was conducted. On the spot urine and salt samples were collected. The Total Goiter Rate was found to be 23.4% and median urinary iodine excretion was 175μg/L. The population is possibly in transition phase from iodine deficient to iodine sufficient nutrition.