Ascorbate-enhanced chondrogenesis of ATDC5 cells

The ATDC5 cell line exhibits the multistep chondrogenic differentiation observed during endochondral bone formation. However, it takes up to two months to complete the process of cell expansion, insulin addition to promote differentiation and further changes in culture conditions effectively to induce hypertrophy. We sought to produce consistent chondrogenesis with significant hypertrophic differentiation with simpler conditions in a more practical time period. By adding ascorbate, the prechondrogenic proliferation phase was shortened from 21 to 7 days, with production of cartilaginous nodules during the chondrogenic phase, after insulin addition, that were greater in number and larger in size. Immunohistochemistry indicated much greater matrix elaboration and the mRNA expression of sox9, aggrecan and collagen type II were all significantly increased earlier and to a much higher degree when compared with controls. Moreover, there was a robust induction of hypertrophy: Col10a1, Runx2 and Mmp13 were all induced within 7-10 days. In conclusion, addition of ascorbate to ATDC5 cultures shortened the prechondrogenic proliferation phase, produced earlier chondrogenic differentiation, heightened gene expression and robust hypertrophic differentiation, abrogating the need for extended culture times and the changes in culture conditions. This simple modification considerably enhances the practicality of this cell line for studies of chondrogenesis

Comparative assessment of haematological profile in hatchery and riverine populations of Channa marulius

798-803Present study was conducted to assess the haematological profile of Channa marulius from hatchery and riverine populations. Blood samples were collected by caudal vein puncture. Significantly higher values (p ≤ 0.05) were recorded for various blood indices in hatchery populations as compared to those of riverine sources. Observed values of haematological parameters in hatchery populations were: Erythrocytes 4.90×106 cells µL-1, eosinophils 4.06 %, monocytes 5.53 %, haematocrit count 28.86 %, haemoglobin content 5.385 gdL-1, platelet distribution width 6.91 fL, red blood cell distribution width 23.83 fL, Procalcitonin 2.37 µL, mean corpuscular volume 7.87 fL and large platelet concentration ratio 14.64 %. In riverine populations, significantly higher values (p ≤ 0.05) for leucocytes count 5625×103 µL-1, mean cell haemoglobin 32.75 pg, mean corpuscular haemoglobin concentration 43.66 gdL-1 and mean cell volume 171.90 fL were observed as compared to those of hatchery populations. Conversely, non-significant (p ≥ 0.05) differences were observed with elevated values for neutrophils (14.042 %) in riverine fish populations as compared hatchery samples (13.896 %). Lymphocytes and platelets counts were 17.344 % and 33.742×103 µL-1, respectively in hatchery populations whereas in riverine populations these were 13.764 % and 33.896×103 µL-1, respectively. Physico-chemical parameters of sample water were observed to be in safe range throughout the study period. The observed variation in haematological profile between both groups is due to different inhabiting conditions that exert direct impacts on fish haematology

A phase I open-label, dose-escalation study of NUC-3373, a targeted thymidylate synthase inhibitor, in patients with advanced cancer (NuTide:301)

\ua9 The Author(s) 2024.Purpose: 5-fluorouracil (5-FU) is inefficiently converted to the active anti-cancer metabolite, fluorodeoxyuridine-monophosphate (FUDR-MP), is associated with dose-limiting toxicities and challenging administration schedules. NUC-3373 is a phosphoramidate nucleotide analog of fluorodeoxyuridine (FUDR) designed to overcome these limitations and replace fluoropyrimidines such as 5-FU. Patients and methods: NUC-3373 was administered as monotherapy to patients with advanced solid tumors refractory to standard therapy via intravenous infusion either on Days 1, 8, 15 and 22 (Part 1) or on Days 1 and 15 (Part 2) of 28-day cycles until disease progression or unacceptable toxicity. Primary objectives were maximum tolerated dose (MTD) and recommended Phase II dose (RP2D) and schedule of NUC-3373. Secondary objectives included pharmacokinetics (PK), and anti-tumor activity. Results: Fifty-nine patients received weekly NUC-3373 in 9 cohorts in Part 1 (n = 43) and 3 alternate-weekly dosing cohorts in Part 2 (n = 16). They had received a median of 3 prior lines of treatment (range: 0–11) and 74% were exposed to prior fluoropyrimidines. Four experienced dose-limiting toxicities: two Grade (G) 3 transaminitis; one G2 headache; and one G3 transient hypotension. Commonest treatment-related G3 adverse event of raised transaminases occurred in < 10% of patients. NUC-3373 showed a favorable PK profile, with dose-proportionality and a prolonged half-life compared to 5-FU. A best overall response of stable disease was observed, with prolonged progression-free survival. Conclusion: NUC-3373 was well-tolerated in a heavily pre-treated solid tumor patient population, including those who had relapsed on prior 5-FU. The MTD and RP2D was defined as 2500 mg/m2 NUC-3373 weekly. NUC-3373 is currently in combination treatment studies. Trial registration: Clinicaltrials.gov registry number NCT02723240. Trial registered on 8th December 2015. https://clinicaltrials.gov/study/NCT02723240

Comparative assessment of serum biochemical profile in riverine and cultured populations of Channa marulius (Hamilton, 1822)

1010-1016Serum biochemical assessment is an important tool to provide information about the internal organs, metabolic and nutritional parameters and electrolytes. Present study was conducted to determine the serum biochemistry of Channa marulius in hatchery and riverine populations. For this purpose blood samples were collected by caudal vein puncture. Inferences of the study showed non-significantly (p ≥ 0.05) higher values of calcium 10.24 mmol/L as compared to lower 9.66 mmol/L whereas, significantly (p ≤ 0.05) higher values of serum globulin 21.06 g/L, total protein 53.0 g/L, albumin 14.87 g/L, cholesterol 6.116 mmol/L, alanine transaminase 530 U/L and alkaline phosphatase 398 µ/L were observed in cultured populations as compared to lower 20.40 g/L, 44.84 g/L, 12.30 g/L, 5.084 mmol/L, 453 U/L and 255 µ/L in riverine populations. On the other side, non-significantly (p ≥ 0.05) higher values of serum triglyceride 0.674 mmol/L as compared to lower 0.604 mmol/L, while significantly (p ≤ 0.05) higher values of glucose 28.43 g/L, urea 2.541 mmol/L, bilirubin 28.20 µmol/L, chlorine 70.3 mmol/L, potassium 13.43 mmol/L, sodium 126.5 mmol/L and aspartate transaminase 1169 U/L in riverine populations as compared to lower 23.89 g/L, 2.17 mmol/L, 26.23 µmol/L, 61.9 mmol/L, 12.08 mmol/L, 111.2 mmol/L and 1029 U/L were recorded in cultured populations, respectively. Findings of the study will be helpful in the field of biochemistry, physiology and toxicology as well as to enhance the management and rearing potential of the Chnanna marulius

Haptoglobin genotype and outcome after spontaneous intracerebral haemorrhage

OBJECTIVE: Haptoglobin is a haemoglobin-scavenging protein that binds and neutralises free haemoglobin and modulates inflammation and endothelial progenitor cell function. A HP gene copy number variation (CNV) generates HP1 and HP2 alleles, while the single-nucleotide polymorphism rs2000999 influences their levels. The HP1 allele is hypothesised to improve outcome after spontaneous (non-traumatic) intracerebral haemorrhage (ICH). We investigated the associations of the HP CNV genotype and rs2000999 with haematoma volume, perihaematomal oedema (PHO) volume, functional outcome and mortality after ICH. METHODS: We included patients with neuroimaging-proven ICH, available DNA and 6-month follow-up in an observational cohort study (CROMIS-2). We classified patients into three groups according to the HP CNV: 1-1, 2-1 or 2-2 and also dichotomised HP into HP1-containing genotypes (HP1-1 and HP2-1) and HP2-2 to evaluate the HP1 allele. We measured ICH and PHO volume on CT; PHO was measured by oedema extension distance. Functional outcome was assessed by modified Rankin score (unfavourable outcome defined as mRS 3-6). RESULTS: We included 731 patients (mean age 73.4, 43.5% female). Distribution of HP CNV genotype was: HP1-1 n=132 (18.1%); HP2-1 n=342 (46.8%); and HP2-2 n=257 (35.2%). In the multivariable model mortality comparisons between HP groups, HP2-2 as reference, were as follows: OR HP1-1 0.73, 95% CI 0.34 to 1.56 (p value=0.41) and OR HP2-1 0.5, 95% CI 0.28 to 0.89 (p value=0.02) (overall p value=0.06). We found no evidence of association of HP CNV or rs200999 with functional outcome, ICH volume or PHO volume. CONCLUSION: The HP2-1 genotype might be associated with lower 6-month mortality after ICH; this finding merits further study

Acquisition of the Sda1-encoding bacteriophage does not enhance virulence of the serotype M1 Streptococcus pyogenes strain SF370

The resurgence of invasive disease caused by Streptococcus pyogenes (group A Streptococcus [GAS]) in the past 30 years has paralleled the emergence and global dissemination of the highly virulent M1T1 clone. The GAS M1T1 clone has diverged from the ancestral M1 serotype by horizontal acquisition of two unique bacteriophages, encoding the potent DNase Sda1/SdaD2 and the superantigen SpeA, respectively. The phage-encoded DNase promotes escape from neutrophil extracellular traps and is linked to enhanced virulence of the M1T1 clone. In this study, we successfully used in vitro lysogenic conversion to transfer the Sda1-encoding phage from the M1T1 clonal strain 5448 to the nonclonal M1 isolate SF370 and determined the impact of this horizontal gene transfer event on virulence. Although Sda1 was expressed in SF370 lysogens, no capacity of the phage-converted strain to survive human neutrophil killing, switch to a hyperinvasive covRS mutant form, or cause invasive lethal infection in a humanized plasminogen mouse model was observed. This work suggests that the hypervirulence of the M1T1 clone is due to the unique synergic effect of the M1T1 clone bacteriophage-specific virulence factor Sda1 acting in concert with the M1T1 clone-specific genetic scaffold

Production and utilization aspects of waste cooking oil based biodiesel in Pakistan

Excessive fuel demand thrusts the Pakistani government to import large volumes of fuel from foreign sources, creating adverse effects on the country's economy. Therefore, exploring an alternative to fossil fuels is unavoidable. The option of environmentally friendly fuel like biodiesel produced from indigenous waste is an additional bonus for the populous developing country like Pakistan where likelihood of waste generation is huge. There exists a potential option for sustainable biodiesel production utilizing excessive waste cooking oil available in the country which otherwise is an ecological burden. The present work is focused to sturdily vindicate the appropriateness of waste cooking oil-based biodiesel generation and utilization in Pakistan through SWOT-AHP, TOWS and PESTLE analysis. The prioritization of SWOT through AHP in view of experts’ perception displayed the strengths and opportunities in highest group priority values (Strengths: 0.51, Opportunities: 0.29). Furthermore, TOWS analysis suggests promising strategies for the sustainable implementation of commercial aspect of waste oil-based biodiesel in Pakistan. Political, Economic, Social, Technological, Legal and Environmental (PESTLE) analysis favors the strengths and opportunities factors of SWOT and TOWS strategies for the application of waste cooking oil based biodiesel in country. At the end, regional recommendations have been provided for the implementation of biodiesel production scenario in country

Partial characterization of glutathione S-transferase (GST) isolated from hepatocytes of Cyprinus carpio from river Ravi Punjab, Pakistan

1017-1024Glutathione S-transferase (GST) is vital in oxidative stress. Keeping in view the importance of this enzyme, present study was conducted for partial characterization of GST from hepatocytes of Cyprinus carpio. The inferences showed higher (GST) activity in liver of C. carpio collected from Chashma Barrage (144.33±1.15) as compared to Trimmu Head works (128.66.3±0.577) and Rasul Barrage (111.66±0.577) sampling sites. The highest GST activity was 128±0.333 U mL-1 from the crude extract fraction of liver as compared to other fractions of appraisal fish liver. Whereas the lowest activity (109±0.666 U mL-1) was from desalted fraction. Optical density was recorded by spectrophotometer at 280 nm, the fractions that showed high optical density were subjected to enzyme assay. Highest specific activity was 183.63, 153.84 and 162.50 U mg-1 for purified liver GST of C. carpio captured from Trimmu Head works, Rasul Barrage and Chashma Barrage, respectively. Whereas, fold purification was 2.86, 2.69 and 2.10 while, percent recovery was 78.90, 72.07 and 63.19 %, respectively for purified liver GST. The optimum pH was 6.5, 6.0 and 6.8 whereas, temperature was 30.5 oC, 31 oC and 30 oC followed by substrate concentration 40.3 mM, 40 mM and 41 mM, for purified liver GST of C. carpio captured from Trimmu Head works, Rasul Barrage and Chashma Barrage, respectively. It should be noted that detailed studies can be beneficial in the fields of physiology, biochemistry and toxicology