WEE1 kinase inhibitor MK-1775 sensitizes oral tongue squamous cell carcinoma cells to radiation irrespective of TP53 status.

OBJECTIVE: Oral tongue squamous cell carcinoma (OTSCC) frequently harbors non-functional p53 and depends on G2/M checkpoint mediated by WEE1. WEE1 suppression has been identified as a promising anti-tumor strategy. This study investigated the capacity of WEE1 kinase inhibitor (MK-1775) and its underlying mechanisms in enhancing radiation responses of OTSCC cells in vitro. MATERIALS AND METHODS: WEE1 kinase expression and its downstream target (CDK1) were investigated in OTSCC versus normal oral tissue. A synergistic combination of MK-1775 with radiation on OTSCC cell lines with different p53 statuses was assessed by viability assay. The radio-sensitizing effects of MK-1775 on apoptosis, cell cycle, DNA damage, and mitotic entry were also determined. RESULTS: Irradiation enhanced CDK1 expression in all tested cell lines, though the effect was far more pronounced in p53 mutated cell lines. MK-1775 exhibited inhibitory effects against the survival of all cell lines and enhanced their response to the radiation. These effects were strongly elicited by induction of apoptosis and lethal mitosis, but less likely by abrogation of radiation-induced G2 arrest. CONCLUSION: These results demonstrate the efficacy of MK-1775 in enhancing the radiation effect on OTSCC in vitro associated with a significant apoptotic death rate, identifying WEE1 inhibitor as a potent radiosensitizer in OTSCC irrespective of p53 mutational status.

Association of haematological biomarkers with severity of COVID-19 pneumonia.

BACKGROUND: Coronavirus disease 2019 (COVID-19) was first reported in Wuhan, China in December 2019. It is caused by SARS-CoV-2, a beta coronavirus. In this study, we assessed the association of biomarkers such as neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR) and lymphocyte monocyte ratio (LMR) with the severity of COVID-19 in patients. METHODS: This retrospective observational study was carried out at a tertiary care hospital of the sub-Himalayan region of Uttarakhand over a period of six months from May to October 2020. A total of 350 patients with confirmed RT-PCR COVID-19 infection were included in the study. Detailed clinical, demographic and biochemical data of each patient was obtained from the hospital record section after permission from the Institute Ethical Committee. NLR, PLR and LMR ratios were calculated and compared with the outcomes in each patient. The patients were subdivided into two sub-groups: those with saturation less than 94% and those with saturation more than 94%. The patients were categorised as mild (with SpO2 of > 94%) and moderate-severe (with SpO2 of ≤94%) based on oxygen saturation. RESULTS: A total of 350 patients with Covid-19 pneumonia were enrolled in the study. The mean age of the patients with oxygen saturation of ≤94% was 54.91 ± 13.29 years, which was comparable to the other group. Absolute neutrophil count (ANC) and NLR were significantly higher in patients with a saturation of < 94%. However, LMR and PLR were significantly lower in the group with saturation of <94%. Thus, a significant association was found between haematological inflammatory ratios and the severity of COVID-19 infection. CONCLUSION: NLR, LMR and PLR ratios can be utilised as point of care markers to assess severity in patients with COVID-19 pneumonia.

Angiotoxicity of oxygenated sterols and possible precursors.

Cell death, inflammation, and repair in rabbits' aortas and pulmonary arteries were observed at 3-, 7-, and 10-day periods after the intravenous injection of oxygenated sterols. Thus, oxygenated sterols, not cholesterol, may play the primary role in arterial wall injury and lesion development.

Chemoimmunotherapy of cancer.

The preparation of a series of water-soluble mustard haptens for chemoimmunotherapy of cancer is described. Preliminary screening data are given, indicating some activity against P388 lymphocytic leukemia for those compounds containing the most potent immunogenic functional groups.

Chemoimmunotherapy of cancer. 3. Analytical measurement of chemical half-lives of monofunctional alkylators.

The objective of this study is the measurement of the rates of hydrolysis of a series of chloroethyl sulfide derivatives, under stimulated physiological conditions. Interferences encountered with the conventional spectrophotometric method prompted the use of a rapid-response, chloride selective electrode. This probe was readily capable of monitoring the hydrolytic rate, which is identical with the rate of chloride ion formation. Since the desired subsecond half-lives were not achieved by any of the compounds, factors influencing the rates were investigated. The results suggest that the rate-controlling cyclization step may be inhibited, due to coordination of undissociated protonic functional groups on the aromatic portions of the structures with the lone-pair electrons on sulfur.

New brain perfusion imaging agents based on 99mTc-bis(aminoethanethiol) complexes: stereoisomers and biodistribution.

In developing new brain perfusion imaging agents, we prepared 99mTc complexes of racemic mixtures of bis(aminoethanethiol) (BAT) derivatives containing an N'-benzylpiperazinyl (BPA) side chain. Due to the presence of a chiral center, a mixture of diastereomers (syn and anti) following chelation with the 99mTc (no-carrier-added) was obtained. The neutral and lipid-soluble 99mTc-BPA-BAT (99mTc, T1/2 = 6 h) isomers were separated. The syn and anti isomers of carrier-added 99Tc-BPA-BAT (99Tc, T1/2 = 2 x 10(5) years) were also synthesized, separated, and crystallized. The X-ray crystallography of 99Tc-BPA-BAT showed the syn and anti conformations (in relationship with the central TC(=O)N2S2 core). Despite a similarity in the partition coefficients for the two isomers, the syn isomer showed a higher in vivo brain uptake and longer brain retention in rats (2.77 and 1.08% dose/organ at 2 and 15 min) than that of the corresponding anti isomer (0.57 and 0.27% dose/organ at 2 and 15 min). This information is important and should be taken into consideration when new 99mTc-labeled brain perfusion imaging agents are being designed.

Management of pulmonary tuberculosis suspects with negative sputum smears and normal or minimally abnormal chest radiographs in resource-poor settings.

SETTING: Queen Elizabeth Central Hospital, Blantyre, Malawi. OBJECTIVES: 1) To determine the proportion of pulmonary tuberculosis (PTB) suspects with negative sputum smears and a normal/minimally abnormal chest radiograph (CXR) who are culture-positive for Mycobacterium tuberculosis, and 2) to determine how many develop smear or radiographic evidence of PTB (TB CXR) during follow-up. METHODS: PTB suspects with negative sputum smears and a normal/minimally abnormal CXR were given a second course of antibiotics and followed up at 3-week intervals over 3 months with repeat sputum smears and chest radiography. RESULTS: Of 79 patients (38 men and 41 women, mean age 33 years) with negative smears and a normal/minimally abnormal CXR, 16 (21%) were culture-positive for M. tuberculosis. Of 15 culture-positive patients who were alive and attended follow-up, seven (47%) developed a TB-CXR by 3 months. Of 41 culture-negative patients who were alive and attended follow-up, 13 (32%) developed a TB-CXR, including one patient who became sputum smear-positive. TB-CXRs were found only in patients with a cough. CONCLUSION: TB suspects with negative smears and normal/minimally abnormal CXRs in high human immunodeficiency virus (HIV) prevalent countries should be given a second course of antibiotics. If cough improves, patients can be advised not to return for further follow-up. If cough continues, patients should return for further follow-up with sputum smear examination and chest radiography. Approximately 50% of those who have culture-positive PTB will develop a TB-CXR by 3 months and can be identified if radiographic facilities are available.

Efficacy of an unsupervised ambulatory treatment regimen for smear-negative pulmonary tuberculosis and tuberculous pleural effusion in Malawi.

SETTING: Queen Elizabeth Central Hospital, Blantyre, and Zomba Central Hospital, Zomba, Malawi. OBJECTIVE: To evaluate treatment outcome of unsupervised ambulatory treatment (2R3H3Z3/2TH[EH]/4H) in Blantyre and 'standard' treatment (1STH[SEH]/11TH[EH]) in Zomba in human immunodeficiency virus (HIV) seropositive and seronegative patients with smear-negative pulmonary tuberculosis (PTB) and pleural TB. DESIGN: All patients with smear-negative and pleural TB registered between 1 April and 31 December 1995 were assessed for enrolment in the study. Study patients were followed up and 12-month treatment outcomes were recorded. RESULTS: A total of 434 patients, 296 with smear-negative PTB and 138 with pleural TB, were enrolled: 366 (84%) of patients were HIV-positive; 220 (51%) completed treatment, and 144 (33%) died by 12 months. In patients from Blantyre and Zomba, baseline characteristics were similar, apart from older age in those from Zomba, and the proportion of patients who completed treatment and who died were similar. In both sites, significantly higher case fatality rates were found in older patients, HIV-positive patients and patients with pulmonary parenchymal lung disease. CONCLUSION: Unsupervised ambulatory treatment evaluated in this study had an efficacy similar to that of 'standard' treatment. For operational reasons, however, it will not be recommended for widespread use in Malawi's National Tuberculosis Control Programme.

Prevalence of tuberculosis in TB suspects with short duration of cough.

The prevalence of pulmonary tuberculosis (PTB) in patients with short duration of cough was determined. Ninety-eight adult out-patients (60 men, 38 women; mean age 32 years) at Queen Elizabeth Central Hospital, Blantyre, Malawi, who had cough for 1-3 weeks which was unresponsive to a course of antibiotics, were successfully screened by microscopy and culture of 2 or 3 sputum specimens and chest radiography; 34 (35%) had PTB. Ten patients were sputum smear-positive and 24 were smear-negative and culture-positive. There was no difference in age, gender or clinical features of general illness, respiratory disease and HIV-related disease between patients with PTB and those with no evidence of PTB. Nine patients (26%) with microbiologically confirmed tuberculosis (TB) had chest radiograph abnormalities consistent with TB, compared with 5 (8%) of patients with no microbiological evidence of TB. Certain classes of patients with a short history of cough would benefit from PTB screening strategies with the emphasis on sputum examination rather than chest radiography, which is unreliable in such patients. The classes include (i) patients with other features of TB whose cough has not improved with antibiotic therapy, (ii) seriously ill patients, and (iii) patients in high risk institutions such as prisons and refugee camps.

Screening pulmonary tuberculosis suspects in Malawi: testing different strategies.

Alternative strategies for screening tuberculosis (TB) suspects are needed in sub-saharan Africa. Ambulatory adult TB suspects who were seen in the chronic cough room of Queen Elizabeth Central Hospital, Blantyre, Malawi, were assessed with respect to appropriateness of referral. Appropriate referrals (patients with cough 3 weeks or longer, weight loss and no antibiotic response) were screened by 3 sputum specimens for microscopy and culture of Mycobacterium tuberculosis and chest radiography (CXR). Hypothetical strategy A (screening by sputum smear examination followed by CXR in patients with negative sputum smears) was compared with strategy B (screening by CXR followed by sputum smear examination in patients with a CXR consistent with TB) in terms of diagnostic efficacy and cost. Of 1127 patients referred to the cough room, 402 (38%) were appropriate TB suspect referrals. Of these, 111 (28%) were sputum smear-positive, 213 (53%) were culture-positive, and 221 (55%) had smear and/or culture-positive evidence of TB. Routine CXR was consistent with pulmonary (P) TB in 230 patients (57%). With strategy A, 243 (60%) patients were diagnosed as PTB, but 40 (25%) of those not diagnosed as PTB had positive mycobacterial cultures. With strategy B, 230 patients (57%) were diagnosed as PTB, but 53 (31%) of those not diagnosed as PTB had positive mycobacterial cultures, including 13 with smear-positive sputum. The cost per diagnosed case of PTB was US$ 4.63 with strategy A and US$ 5.44 with strategy B. Screening patients with good criteria of TB has high diagnostic sensitivity, but screening by CXR is less effective and more costly than screening by sputum smear microscopy.

Tuberculous lymphadenitis, a diagnostic problem in areas of high prevalence of HIV and tuberculosis.

The human immunodeficiency virus (HIV) epidemic is associated with a marked increase of tuberculosis cases. The influence of HIV on diagnostic methods for tuberculous lymphadenitis is less clear. In an environment of high HIV and tuberculosis prevalence in Blantyre, Malawi, a prospective study compared results of basic procedures diagnosing tuberculous lymphadenitis with the outcome of histology and/or culture. One hundred out-patients, aged 15-55 years, with extra-inguinal lymphadenopathy not responding to general antibiotics, entered the study. Among 52 cases, with whom all procedures were carried out in accordance with the protocol, 38 (73%) were diagnosed as tuberculous lymphadenitis; 84% of the latter (32/38) were seropositive for HIV. Needle aspirate and biopsy smears stained by the Ziehl-Neelsen technique contributed little to detecting tuberculosis, 8% and 11% respectively. In contrast, macroscopic caseation of excised lymph nodes showed a high yield of 82%, which was similar to histology, and higher than that of Löwenstein-Jensen culture (61%). The study suggested that HIV positivity of tuberculous lymphadenitis patients decreased the possibility of histology and culture both being indicative of tuberculosis (odds ratio 0.10; P = 0.06). Consequently histology results, often used as the single definitive method, failed to diagnose 18% (7/38) of tuberculosis cases. However, it was reassuring that 4 simple methods, which can safely be carried out at district level, could be expected to diagnose 80-95% of tuberculous lymphadenitis cases in a timely and cost-effective manner.

PIP: A prospective study conducted in 1994-95 in Blantyre, Malawi, investigated the influence of HIV on diagnostic methods for tuberculous lymphadenitis in 100 outpatients 15-55 years old with extra-inguinal lymphadenopathy not responsive to general antibiotics. For the 52 cases for which all diagnostic procedures were carried out, 38 (73%) were diagnosed as tuberculous lymphadenitis; 32 (84%) of these patients were HIV-positive. Needle aspirate and biopsy smears stained by the Ziehl-Neelsen technique contributed only 8% and 11%, respectively, to detecting tuberculosis. In contrast, macroscopic caseation of excised lymph nodes, histology, and Lowenstein-Jensen culture had yields of 82%, 82%, and 61%, respectively. These findings indicate that, in tuberculous lymphadenitis patients, HIV infection decreases the diagnostic power of culture and histology. However, use of the other methods can diagnose 80-95% of tuberculous lymphadenitis cases in a cost-effective manner in areas with high prevalences of both HIV and tuberculosis.

Filariasis and malaria sera from Orissa lack HTLV-III antibody reactivity.

Reports exist indicating a correlation between seropositivity for human T-lymphotrophic virus (HTLV) antibodies and certain parasitic infections in some parts of the world. In 274 filariasis and 119 malaria sera examined from Orissa, none was reactive in a test for anti-HTLV-III antibodies.