PD1 expression and correlation with its ligands in oral cancer specimens and peripheral blood

ObjectivesThis study analyzed the expression of the PD1 receptor in tumor tissue and peripheral blood of oral squamous cell carcinoma (OSCC) patients, and correlated it with the PD1 ligands PD-L1 and PD-L2. The currently low response rates of checkpoint inhibitor treatment in OSCC could be increased by a better understanding of immune checkpoint biology. Despite evidence in the literature for upregulation of PD1 checkpoint ligands in OSCC tissue, there has been no correlation analysis of the PD1 receptor with its ligands in tissue specimens and peripheral blood of OSCC patients.Materials and methodsAn RT-qPCR analysis of PD1 mRNA expression was performed in oral cancer specimens, healthy mucosa, and corresponding blood samples. A cut-off point (COP) was determined and a chi-square (χ²) test was carried out. PD1 expression was correlated with previously reported PD-L1 and PD-L2 expression values using the Spearman test.ResultsTissue and blood specimens of 48 OSCC patients and 26 healthy individuals were analyzed. PD1 expression in OSCC specimens was significantly increased (p = 0.006) compared with healthy oral mucosa. PD1 overexpression in tissue samples showed a significant association with the presence of malignancy (p = 0.006). PD1 expression in tissue samples showed a significant positive correlation (p < 0.001) with the ligands PD-L1 and PD-L2. In contrast, there was no correlation between PD1 and its ligands in blood samples. However, there was a significant positive correlation (p < 0.001) between the ligands PD-L1 and PD-L2, both in tissue and blood samples.ConclusionsIncreased PD1 expression might be a manifestation of T-cell exhaustion in OSCC specimens, leading to immune tolerance. PD-L1/PD-L2-PD1 interaction may be a major mediator of local immunosuppression in OSCC, requiring advanced multimodal treatment protocols.     

Homogeneous liquid–liquid microextraction based on liquid nitrogen-induced phase separation followed by GFAAS for sensitive extraction and determination of lead in lead-adulterated opium and refined opium

Herein, we developed a novel homogeneous liquid–liquid microextraction based on liquid nitrogen-induced phase separation (HLLME-LNPS) for the extraction and determination of lead (Pb) in Pb-adulterated opium and refined opium by GFAAS analysis. In this procedure, first, 400 μl of acetonitrile (extractant) containing 7.0 μl of diethyl dithiophosphoric acid (DDTP) is injected into a sample solution and a homogeneous solution is formed. Subsequently, the homogeneous mixture is cooled using liquid nitrogen for 16 seconds. By this process, due to the difference in the freezing points of the organic and aqueous phases, the homogeneous state is broken and the Pb-DDTP species are extracted into the liquid organic phase collected on top of the frozen aqueous phase. The introduced method exhibited a good linearity with a coefficient of determination (r²) of 0.9988 and an acceptable linear range of 0.6–100 μg l⁻¹. Accordingly, the detection limit was 0.2 μg l⁻¹ (S/N = 3) for Pb ions, and a high enrichment factor was obtained. The proposed method was successfully utilized to determine trace levels of Pb in opium samples. The results of the sample analysis showed that 65% of the opium samples and 85% of the refined opium samples had much higher than expected levels of contaminating Pb, and this contamination poses a serious threat to drug users.

Herein, we developed a novel homogeneous liquid–liquid microextraction based on liquid nitrogen-induced phase separation (HLLME-LNPS) for the extraction and determination of lead (Pb) in Pb-adulterated opium and refined opium by GFAAS analysis.     

Effect of micro-aerobic process on improvement of anaerobic digestion sewage sludge treatment: flow cytometry and ATP assessment

Micro-aeration as a pretreatment method improves the efficiency of anaerobic digestion of municipal sewage sludge and consequently promotes the methane production. In this study, adenosine triphosphate (ATP) and flow cytometry (FCM) were employed to monitor the performance of the micro-aerobic process and investigate the survival of bacterial cells within the process. At first, the effect of air flow rate (AFR) (0.1, 0.2, 0.3 and 0.5 vvm) on hydrolysis of mixed sludge in 5 aeration cycles (20, 30, 40, 48 and 60 hours) was examined. Then, the effects of the micro aerobic process on methane (CH₄) production in anaerobic digestion were surveyed. The highest VSS reduction was 30.6% and 10.4% for 40 hours in the reactor and control, respectively. Soluble COD also fluctuated between 40.87 and 65.14% in micro-aerobic conditions; the highest SCOD was achieved at the time of 40 h. Microbial activities were increased by 597%, 170% and 79.4% for 20, 30 and 40 h pretreatment with the micro-aerobic process, respectively. Apoptosis assay showed that micro-aerobic pre-treatment at 20, 30 and 40 h increased the percentage of living cells by 57.4, 62.8 and 67.9%, respectively. On the other hand, FCM results showed that the highest percentage of viable bacteria (i.e., 67.9%) was observed at 40 h pretreating which was approximately 40% higher the ones for the control. Variation in cumulative methane production shows that methane production was increased by 221% compared to anaerobic digestion (control group). Therefore, ATP and FCM can be employed as two appropriate, accurate, relatively specific indicators for monitoring the process and bacteria viability.

Micro-aeration as a pretreatment method improves the efficiency of anaerobic digestion of municipal sewage sludge and consequently promotes the methane production.     

How effective is the early fast treadmill gait speed training for stroke patients at the 2nd week after admission: comparison with comfortable gait speed at the 6th week

[Purpose] The purpose of this study was to find whether a fast treadmill gait training speed is effective for the gait training of stroke patients in the early rehabilitation stage. [Subjects and Methods] Thirty-nine stroke patients were the subjects of our investigation. They walked on a treadmill with handrail supports at a fast speed (130% of their comfortable gait speed in the 2nd week). The treadmill gaits of the patients were recorded using a 3-dimensional analysis system at two and six weeks after their admissions. Intraclass Correlation Coefficients (ICC) of the temporal and spatial parameters of the two periods were statistically analyzed. [Results] For all of the patients, the ICCs of the measured parameters were greater than 0.58. In the case of patients whose gait speeds of the two periods were close, the ICC units were greater than 0.7. [Conclusion] The fast gait speed training allowed us to expose the patients to a gait speed that they were expected to acquire at a later stage of their rehabilitation. This training method was found to be beneficial for the mildly paralyzed patients.Key words: Stroke, Treadmill, Fast speed     

Inferior glenohumeral joint dislocation with greater tuberosity avulsion

Inferior glenohumeral dislocation is the least common type of glenohumeral dislocations. It may be associated with fractures of the adjacent bones and neurovascular compromise. It should be treated immediately by close reduction. The associated neuropraxia usually recovers with time. Traction-counter traction method is commonly used for reduction followed by immobilization of the shoulder for three weeks. Here, we report a case of inferior glenohumeral joint dislocation with greater tuberosity fracture with transient neurovascular compromise and present a brief review of the literature.     

First report of neonatal early-onset sepsis caused by multi-drug-resistant <Emphasis Type="Italic">Raoultella ornithinolytica</Emphasis>

Raoultella ornithinolytica is increasingly being isolated as a causative organism in human infections. Most of the infections caused by R. ornithinolytica are hospital acquired and occur in patients who are immunocompromised, had invasive procedures or have indwelling catheters. This is a first report of early onset neonatal sepsis caused by multi-drug-resistant R. ornithinolytica. The infection was not very severe and was characterised by generalized flushing of the skin. Patient made complete recovery once appropriate antibiotics were started.