A comprehensive comparison of machine learning models for ICH prognostication: Retrospective review of 1501 intra-cerebral hemorrhage patients from the Qatar stroke database.

Multiple prognostic scores have been developed to predict morbidity and mortality in patients with spontaneous intracerebral hemorrhage(sICH). Since the advent of machine learning(ML), different ML models have also been developed for sICH prognostication. There is however a need to verify the validity of these ML models in diverse patient populations. We aim to create machine learning models for prognostication purposes in the Qatari population. By incorporating inpatient variables into model development, we aim to leverage more information. 1501 consecutive patients with acute sICH admitted to Hamad General Hospital(HGH) between 2013 and 2023 were included. We trained, evaluated, and compared several ML models to predict 90-day mortality and functional outcomes. For our dataset, we randomly selected 80% patients for model training and 20% for validation and used k-fold cross validation to train our models. The ML workflow included imbalanced class correction and dimensionality reduction in order to evaluate the effect of each. Evaluation metrics such as sensitivity, specificity, F-1 score were calculated for each prognostic model. Mean age was 50.8(SD 13.1) years and 1257(83.7%) were male. Median ICH volume was 7.5 ml(IQR 12.6). 222(14.8%) died while 897(59.7%) achieved good functional outcome at 90 days. For 90-day mortality, random forest(RF) achieved highest AUC(0.906) whereas for 90-day functional outcomes, logistic regression(LR) achieved highest AUC(0.888). Ensembling provided similar results to the best performing models, namely RF and LR, obtaining an AUC of 0.904 for mortality and 0.883 for functional outcomes. Random Forest achieved the highest AUC for 90-day mortality, and LR achieved the highest AUC for 90-day functional outcomes. Comparing ML models, there is minimal difference between their performance. By creating an ensemble of our best performing individual models we maintained maximum accuracy and decreased variance of functional outcome and mortality prediction when compared with individual models.

Oxalate decarboxylase uses electron hole hopping for catalysis.

The hexameric low-pH stress response enzyme oxalate decarboxylase catalyzes the decarboxylation of the oxalate mono-anion in the soil bacterium Bacillus subtilis. A single protein subunit contains two Mn-binding cupin domains, and catalysis depends on Mn(III) at the N-terminal site. The present study suggests a mechanistic function for the C-terminal Mn as an electron hole donor for the N-terminal Mn. The resulting spatial separation of the radical intermediates directs the chemistry toward decarboxylation of the substrate. A π-stacked tryptophan pair (W96/W274) links two neighboring protein subunits together, thus reducing the Mn-to-Mn distance from 25.9 Å (intrasubunit) to 21.5 Å (intersubunit). Here, we used theoretical analysis of electron hole-hopping paths through redox-active sites in the enzyme combined with site-directed mutagenesis and X-ray crystallography to demonstrate that this tryptophan pair supports effective electron hole hopping between the C-terminal Mn of one subunit and the N-terminal Mn of the other subunit through two short hops of ∼8.5 Å. Replacement of W96, W274, or both with phenylalanine led to a large reduction in catalytic efficiency, whereas replacement with tyrosine led to recovery of most of this activity. W96F and W96Y mutants share the wildtype tertiary structure. Two additional hole-hopping networks were identified leading from the Mn ions to the protein surface, potentially protecting the enzyme from high Mn oxidation states during turnover. Our findings strongly suggest that multistep hole-hopping transport between the two Mn ions is required for enzymatic function, adding to the growing examples of proteins that employ aromatic residues as hopping stations.

Expansion of the Family of Molecular Nanoparticles of Cerium Dioxide and Their Catalytic Scavenging of Hydroxyl Radicals.

The syntheses, crystal structures, and catalytic radical scavenging activity are reported for four new molecular clusters that have resulted from a bottom-up molecular approach to nanoscale CeO2. They are [Ce6O4(OH)4(dmb)12(H2O)4] (dmb- = 2,6-dimethoxybenzoate), [Ce16O17(OH)6(O2CPh)24(HO2CPh)4], [Ce19O18(OH)9(O2CPh)27(H2O)(py)3], and [Ce24O27(OH)9(O2CPh)30(py)4]. They represent a major expansion of our family of so-called "molecular nanoparticles" of this metal oxide to seven members, and their crystal structures confirm that their cores all possess the fluorite structure of bulk CeO2. In addition, they have allowed the identification of surface features such as the close location of multiple Ce3+ ions and organic ligand binding modes not seen previously. The ability of all seven members to catalytically scavenge reactive oxygen species has been investigated using HO• radicals, an important test reaction in the ceria nanoparticle biomedical literature, and most have been found to exhibit excellent antioxidant activities compared to traditional ceria nanoparticles, with their activity correlating inversely with their surface Ce3+ content.

An Interprotein Co-S Coordination Complex in the B12-Trafficking Pathway.

The CblC and CblD chaperones are involved in early steps in the cobalamin trafficking pathway. Cobalamin derivatives entering the cytoplasm are converted by CblC to a common cob(II)alamin intermediate via glutathione-dependent alkyltransferase or reductive elimination activities. Cob(II)alamin is subsequently converted to one of two biologically active alkylcobalamins by downstream chaperones. The function of CblD has been elusive although it is known to form a complex with CblC under certain conditions. Here, we report that CblD provides a sulfur ligand to cob(II)alamin bound to CblC, forming an interprotein coordination complex that rapidly oxidizes to thiolato-cob(III)alamin. Cysteine scanning mutagenesis and EPR spectroscopy identified Cys-261 on CblD as the sulfur donor. The unusual interprotein Co-S bond was characterized by X-ray absorption spectroscopy and visualized in the crystal structure of the human CblD thiolato-cob(III)alamin complex. Our study provides insights into how cobalamin coordination chemistry could be utilized for cofactor translocation in the trafficking pathway.

Chlorocob(II)alamin Formation Which Enhances the Thiol Oxidase Activity of the B12-Trafficking Protein CblC.

CblC is a chaperone that catalyzes removal of the ß-axial ligand of cobalamin (or B12), generating cob(II)alamin in an early step in the cofactor trafficking pathway. Cob(II)alamin is subsequently partitioned to support cellular needs for the synthesis of active cobalamin cofactor derivatives. In addition to the ß-ligand transferase activity, the Caenorhabdiitis elegans CblC (ceCblC) and clinical R161G/Q variants of the human protein exhibit robust thiol oxidase activity, converting glutathione to glutathione disulfide while concomitantly reducing O2 to H2O2. The chemical efficiency of the thiol oxidase side reaction during ceCblC-catalyzed dealkylation of alkylcobalamins is noteworthy in that it effectively scrubs ambient oxygen from the reaction mixture, leading to air stabilization of the highly reactive cob(I)alamin product. In this study, we report that the enhanced thiol oxidase activity of ceCblC requires the presence of KCl, which explains how the wasteful thiol oxidase activity is potentially curtailed inside cells where the chloride concentration is low. We have captured an unusual chlorocob(II)alamin intermediate that is formed in the presence of potassium chloride, a common component of the reaction buffer, and have characterized it by electron paramagnetic resonance, magnetic circular dichroism, and computational analyses. The ability to form a chlorocob(II)alamin intermediate could represent an evolutionary vestige in ceCblC, which is structurally related to bacterial B12-dependent reductive dehalogenases that have been proposed to form halogen cob(II)alamin intermediates in their catalytic cycle.

Cofactor Editing by the G-protein Metallochaperone Domain Regulates the Radical B12 Enzyme IcmF.

IcmF is a 5'-deoxyadenosylcobalamin (AdoCbl)-dependent enzyme that catalyzes the carbon skeleton rearrangement of isobutyryl-CoA to butyryl-CoA. It is a bifunctional protein resulting from the fusion of a G-protein chaperone with GTPase activity and the cofactor- and substrate-binding mutase domains with isomerase activity. IcmF is prone to inactivation during catalytic turnover, thus setting up its dependence on a cofactor repair system. Herein, we demonstrate that the GTPase activity of IcmF powers the ejection of the inactive cob(II)alamin cofactor and requires the presence of an acceptor protein, adenosyltransferase, for receiving it. Adenosyltransferase in turn converts cob(II)alamin to AdoCbl in the presence of ATP and a reductant. The repaired cofactor is then reloaded onto IcmF in a GTPase-gated step. The mechanistic details of cofactor loading and offloading from the AdoCbl-dependent IcmF are distinct from those of the better characterized and homologous methylmalonyl-CoA mutase/G-protein chaperone system.

Sacrificial Cobalt-Carbon Bond Homolysis in Coenzyme B12 as a Cofactor Conservation Strategy.

A sophisticated intracellular trafficking pathway in humans is used to tailor vitamin B12 into its active cofactor forms, and to deliver it to two known B12-dependent enzymes. Herein, we report an unexpected strategy for cellular retention of B12, an essential and reactive cofactor. If methylmalonyl-CoA mutase is unavailable to accept the coenzyme B12 product of adenosyltransferase, the latter catalyzes homolytic scission of the cobalt-carbon bond in an unconventional reversal of the nucleophilic displacement reaction that was used to make it. The resulting homolysis product binds more tightly to adenosyltransferase than does coenzyme B12, facilitating cofactor retention. We have trapped, and characterized spectroscopically, an intermediate in which the cobalt-carbon bond is weakened prior to being broken. The physiological relevance of this sacrificial catalytic activity for cofactor retention is supported by the significantly lower coenzyme B12 concentration in patients with dysfunctional methylmalonyl-CoA mutase but normal adenosyltransferase activity.

Redox Cycling, pH Dependence, and Ligand Effects of Mn(III) in Oxalate Decarboxylase from Bacillus subtilis.

This contribution describes electron paramagnetic resonance (EPR) experiments on Mn(III) in oxalate decarboxylase of Bacillus subtilis, an interesting enzyme that catalyzes the redox-neutral dissociation of oxalate into formate and carbon dioxide. Chemical redox cycling provides strong evidence that both Mn centers can be oxidized, although the N-terminal Mn(II) appears to have the lower reduction potential and is most likely the carrier of the +3 oxidation state under moderate oxidative conditions, in agreement with the general view that it represents the active site. Significantly, Mn(III) was observed in untreated OxDC in succinate and acetate buffers, while it could not be directly observed in citrate buffer. Quantitative analysis showed that up to 16% of the EPR-visible Mn is in the +3 oxidation state at low pH in the presence of succinate buffer. The fine structure and hyperfine structure parameters of Mn(III) are affected by small carboxylate ligands that can enter the active site and have been recorded for formate, acetate, and succinate. The results from a previous report [Zhu, W., et al. (2016) Biochemistry 55, 429-434] could therefore be reinterpreted as evidence of formate-bound Mn(III) after the enzyme is allowed to turn over oxalate. The pH dependence of the Mn(III) EPR signal compares very well with that of enzymatic activity, providing strong evidence that the catalytic reaction of oxalate decarboxylase is driven by Mn(III), which is generated in the presence of dioxygen.

Establishment of Reference Intervals for Renal Function Clinical Chemistry Parameters in an Indonesian Tertiary Hospital.

Reference intervals aid clinical decision-making for clinical chemistry values. Laboratory test results are compared to reference intervals to aid in the diagnosis, therapy, and monitoring decisions. Due to the differences in ethnicity, gender, age, and analytical methods, reference intervals (RIs) vary between populations. This study aimed to establish the reference values for renal function tests in targeted populations in Indonesia. This research was conducted with a cross-sectional observational analytic design. The research sample consisted of medical check-up data from health professionals at Dr. Mohammad Hoesin Hospital in Palembang, Indonesia. The Kolmogorov-Smirnov test was used to determine the normality of data distribution.   The RIs were computed using reference limits at the 2.5th and 97.5th percentiles (abnormal distribution) or ±two standard deviations (±2 SD) (normal distribution).  The independent t-test (parametric) or Mann-Whitney test was used to compare the RIs of males and females (non-parametric). Males and females had a significant difference (P<0.001) regarding the values of uric acid, urea, and creatinine parameters, requiring the reference intervals to be separated. The following reference intervals were established: uric acid: 230,78-526,99 mol/L for males and 179,03-415.17 mol/L for females, urea: 2,22-4,99 mmol/L for males and 1,78-4,28 mmol/L for females, and creatinine: 61,01-106,99 mol/L for males and 40,67-77,81 mol/L for females. This study defined gender-specific RIs for three renal function test parameters for the adult population of Palembang, Indonesia. The deployment of population-specific RIs may facilitate better laboratory testing.

Comparison of Liver Function Test Results between Architect C8000 and COBAS C501 Automatic Chemistry Analyzer.

Liver function tests are frequently used to screen liver function, monitor therapy, and determine the severity of liver problems. The present study aimed to assess the consistency of the results of the liver function parameters between the two analyzers, Architect c8000 and Cobas C501. This laboratory-based analytical observational study was conducted in a cross-sectional manner. Sample collection was performed through a consecutive sampling procedure from June to December 2019 in the Clinical Pathology Laboratory, Dr. Mohammad Hoesin General Hospital, Palembang, South Sumatra, Indonesia. The research sample consisted of the liver function examination results of patients, carried out using the Architect c8000 and Roche Cobas c501 chemistry analyzers. Serum albumin, alanine transaminase, aspartate aminotransferase, and total protein were the studied variables. The Spearman, Mann-Whitney, and Bland-Altman tests were used to evaluate the comparison test. In total, 100 blood samples were obtained in this study. The results revealed a highly significant correlation (r>0.90, P=0<001) among the four liver function parameters. The results of the liver function parameters inspected by the two analyzers did not differ significantly (P>0.05). In addition, there was a solid agreement on all parameters, with a near-perfect level (concordance correlation coefficient>0.90) and more than 95% of data points falling within the acceptable range. The Architect c8000 and Cobas c501 analyzers produced similar results for liver function tests; hence, these devices can be used interchangeably.

Unravelling the dynamics of Lassa fever transmission with differential infectivity: Modeling analysis and control strategies.

Epidemic models have been broadly used to comprehend the dynamic behaviour of emerging and re-emerging infectious diseases, predict future trends, and assess intervention strategies. The symptomatic and asymptomatic features and environmental factors for Lassa fever (LF) transmission illustrate the need for sophisticated epidemic models to capture more vital dynamics and forecast trends of LF outbreaks within countries or sub-regions on various geographic scales. This study proposes a dynamic model to examine the transmission of LF infection, a deadly disease transmitted mainly by rodents through environment. We extend prior LF models by including an infectious stage to mild and severe as well as incorporating environmental contributions from infected humans and rodents. For model calibration and prediction, we show that the model fits well with the LF scenario in Nigeria and yields remarkable prediction results. Rigorous mathematical computation divulges that the model comprises two equilibria. That is disease-free equilibrium, which is locally-asymptotically stable (LAS) when the basic reproduction number, $ {\mathcal{R}}_{0} $, is $ < 1 $; and endemic equilibrium, which is globally-asymptotically stable (GAS) when $ {\mathcal{R}}_{0} $ is $ > 1 $. We use time-dependent control strategy by employing Pontryagin's Maximum Principle to derive conditions for optimal LF control. Furthermore, a partial rank correlation coefficient is adopted for the sensitivity analysis to obtain the model's top rank parameters requiring precise attention for efficacious LF prevention and control.

The dependence of the electrophoretic mobility of small organic ions on ionic strength and complex formation.

The ionic strength dependence of the electrophoretic mobility of small organic anions with valencies up to -3 is investigated in this study. Provided the anions are not too aspherical, it is argued that shape and charge distribution have little influence on mobility. To a good approximation, the electrophoretic mobility of a small particle should be equal to that of a model sphere with the same hydrodynamic radius and same net charge. For small ions, the relaxation effect (distortion of the ion atmosphere from equilibrium due to external electric and flow fields) is significant even for monovalent ions. Alternative procedures of accounting for the relaxation effect are examined. In order to account for the ionic strength dependence of a specific set of nonaromatic and aromatic anions in aqueous solution, it is necessary to include complex formation between the anion with species in the BGE. A number of possible complexes are considered. When the BGE is Tris-acetate, the most important of these involves the complex formed between anion and Tris, the principle cation in the BGE. When the BGE is sodium borate, an anion-anion (borate) complex appears to be important, at least when the organic anion is monovalent. An algorithm is developed to analyze the ionic strength dependence of the electrophoretic mobility. This algorithm is applied to two sets of organic anions from two independent research groups.

Estimation of exponential growth rate and basic reproduction number of the coronavirus disease 2019 (COVID-19) in Africa.

BACKGROUND: Since the first case of coronavirus disease 2019 (COVID-19) in Africa was detected on February 14, 2020, the cumulative confirmations reached 15 207 including 831 deaths by April 13, 2020. Africa has been described as one of the most vulnerable region with the COVID-19 infection during the initial phase of the outbreak, due to the fact that Africa is a great commercial partner of China and some other EU and American countries. Which result in large volume of travels by traders to the region more frequently and causing African countries face even bigger health threat during the COVID-19 pandemic. Furthermore, the fact that the control and management of COVID-19 pandemic rely heavily on a country's health care system, and on average Africa has poor health care system which make it more vulnerable indicating a need for timely intervention to curtail the spread. In this paper, we estimate the exponential growth rate and basic reproduction number (R0) of COVID-19 in Africa to show the potential of the virus to spread, and reveal the importance of sustaining stringent health measures to control the disease in Africa. METHODS: We analyzed the initial phase of the epidemic of COVID-19 in Africa between 1 March and 13 April 2020, by using the simple exponential growth model. We examined the publicly available materials published by the WHO situation report to show the potential of COVID-19 to spread without sustaining strict health measures. The Poisson likelihood framework is adopted for data fitting and parameter estimation. We modelled the distribution of COVID-19 generation interval (GI) as Gamma distributions with a mean of 4.7 days and standard deviation of 2.9 days estimated from previous work, and compute the basic reproduction number. RESULTS: We estimated the exponential growth rate as 0.22 per day (95% CI: 0.20-0.24), and the basic reproduction number, R0, as 2.37 (95% CI: 2.22-2.51) based on the assumption that the exponential growth starting from 1 March 2020. With an R0 at 2.37, we quantified the instantaneous transmissibility of the outbreak by the time-varying effective reproductive number to show the potential of COVID-19 to spread across African region. CONCLUSIONS: The initial growth of COVID-19 cases in Africa was rapid and showed large variations across countries. Our estimates should be useful in preparedness planning against further spread of the COVID-19 epidemic in Africa.

Factors influencing the occurrence of multicentric and 'recurrent' Warthin's tumour: a cross sectional study.

The multicentric nature of Warthin's tumour has been well recognised, though the factors predicting its occurrence and its influence on management remain speculative. In this cross sectional study, the authors analysed the presentation, management and outcome of solitary and multicentric Warthin's tumour, treated in the maxillofacial unit and investigated factors that could influence the occurrence of multicentric and recurrent tumours. Warthin's tumour was found in 24% (150/628) of patients presenting with parotid neoplasms and multicentric tumours were found in 13% (21/ 161) of parotidectomy specimens. Age, sex, side, site and smoking history were not predictors of multicentricity (P>0.40). Clinical examination (19%), imaging investigations (28%) and intra-operative palpation (33%) were poor at detecting multicentric tumours. Superficial parotidectomy was the most commonly performed operation. The recurrence rate was 0% in the solitary and 10% in the multicentric tumour group. Intra-operative rupture was not uncommon (11%), but was not associated with tumour recurrence (P= or >0.999). Incomplete excision (P=0.007) and multicentricity (P=0.026) were predictors of recurrence.

Molecular Rationale for Improved Dynamic Nuclear Polarization of Biomembranes.

Dynamic nuclear polarization (DNP) enhanced solid-state NMR can provide orders of magnitude in signal enhancement. One of the most important aspects of obtaining efficient DNP enhancements is the optimization of the paramagnetic polarization agents used. To date, the most utilized polarization agents are nitroxide biradicals. However, the efficiency of these polarization agents is diminished when used with samples other than small molecule model compounds. We recently demonstrated the effectiveness of nitroxide labeled lipids as polarization agents for lipids and a membrane embedded peptide. Here, we systematically characterize, via electron paramagnetic (EPR), the dynamics of and the dipolar couplings between nitroxide labeled lipids under conditions relevant to DNP applications. Complemented by DNP enhanced solid-state NMR measurements at 600 MHz/395 GHz, a molecular rationale for the efficiency of nitroxide labeled lipids as DNP polarization agents is developed. Specifically, optimal DNP enhancements are obtained when the nitroxide moiety is attached to the lipid choline headgroup and local nitroxide concentrations yield an average e(-)-e(-) dipolar coupling of 47 MHz. On the basis of these measurements, we propose a framework for development of DNP polarization agents optimal for membrane protein structure determination.

Observation of superoxide production during catalysis of Bacillus subtilis oxalate decarboxylase at pH 4.

This contribution describes the trapping of the hydroperoxyl radical at a pH of 4 during turnover of wild-type oxalate decarboxylase and its T165V mutant using the spin-trap BMPO. Radicals were detected and identified by a combination of EPR and mass spectrometry. Superoxide, or its conjugate acid, the hydroperoxyl radical, is expected as an intermediate in the decarboxylation and oxidation reactions of the oxalate monoanion, both of which are promoted by oxalate decarboxylase. Another intermediate, the carbon dioxide radical anion was also observed. The quantitative yields of superoxide trapping are similar in the wild type and the mutant while it is significantly different for the trapping of the carbon dioxide radical anion. This suggests that the two radicals are released from different sites of the protein.

Expression of inducible nitric oxide synthase in bone metastases.

AIMS: The signalling molecule nitric oxide (NO), produced predominantly in cancer by the enzyme inducible NO synthase (iNOS), has been implicated in the pathophysiology of many human tumours. The increased NO concentrations found in many human cancers may facilitate both angiogenesis and tumour dissemination. NO also plays a concentration-dependent role in bone re-modelling by acting on osteoclasts. Although iNOS has been studied extensively in most primary tumours, there are no reports that have investigated its expression in metastatic bone disease. METHODS: An immunohistochemical study was performed using a monoclonal antibody to iNOS in 27 cases of breast, renal and lung bone metastases, biopsied at the time of treatment for pathological fracture. RESULTS: iNOS expression was found in 14 cases and was predominantly localised to tumour cells in the metastatic deposits. A significant difference was found between iNOS expression in metastases and adjacent bone (p<0.001), where immunostaining was rarely seen and confined to immune cells. No microscopic differences in bone architecture could be seen between iNOS positive and negative metastases, and no correlations were found between iNOS expression and clinico-pathological variables. CONCLUSIONS: iNOS expression is not a pathognomonic finding in bone metastasis. Its role in the behaviour of bone metastases requires further investigation.

Expression of type 2 nitric oxide synthase and p21 in oral squamous cell carcinoma.

Nitric oxide (NO) has a complex role in tumour biology. Most cancer research has focused on the enzyme nitric oxide synthase-2 (NOS2), an inducible isoform responsible for prolonged NO production. In normal cells exposed to high NO concentrations, the tumour-suppressor gene, p53, promotes apoptosis via the p21 pathway, in an attempt to safeguard against potential NO-mediated DNA damage. In cancer cells with mutant p53, this pathway is unlikely to occur directly, although, p53-independent p21 expression and subsequent apoptosis can occur at higher NO concentrations. In this study, the possible direct association between NOS2 and p21 was assessed in oral squamous cell carcinoma. Immunohistochemistry was performed for NOS2 and p21 on 56 cases, and NOS2 activity was determined with citrulline assays in selected cases. A significant relationship was demonstrated between the immunohistochemical expression of NOS2 and its activity (P<0.001), but not between NOS2 and p21 expression (P=0.76). It is unlikely that the NO concentrations found in oral cancer (up to 10.3 pmol NO min(-1) mg protein(-1)) are sufficient to cause direct (p53-independent) p21 accumulation and subsequent apoptosis. As with many other tumours, since NO production has a detrimental role, its pharmacological inhibition in oral cancer represents an exciting area for possible future therapeutic manipulation.

Expression of inducible nitric oxide synthase in cutaneous adnexal tumours of the head and neck.

Adnexal tumours are a rare heterogeneous group of neoplasms, most commonly found in the head and neck region. Although most are benign, malignant adnexal tumours often behave aggressively, resulting in early metastasis. There is increasing interest in the role that nitric oxide (NO) plays in the behaviour of many cancers. It is thought that NO, produced by the enzyme inducible NO synthase (iNOS), facilitates tumour growth and dissemination. iNOS has been studied in the common skin cancers, where its expression correlates with tumour behaviour, but it has not been previously investigated in cutaneous adnexal tumours. An immunhistochemical study was performed using a monoclonal antibody to iNOS in 37 cases of adnexal tumours (19 benign, 18 malignant). iNOS expression was weakly expressed by basal keratinocytes of adjacent skin in all cases and it was variably expressed in the tumours. Malignant tumours had significantly increased iNOS expression when compared to both adjacent skin (P<0.001) and the benign tumour group (P<0.001). No significant difference was found between iNOS expression in benign tumour and adjacent skin (P=0.5). The role of iNOS in this rare group of tumours and the possibility of pharmacologically inhibiting it in the clinical setting warrants further investigation.

Expression of nitric oxide synthase-2 in cutaneous squamous cell carcinoma of the head and neck.

Nitric oxide (NO) has a pivotal role in angiogenesis. The expression of NO synthase (NOS2) is down-regulated in basal cell skin cancer but we know of no studies that have investigated its expression in the more aggressive squamous cell skin carcinoma. This immunohistochemical study assessed NOS2 expression in 37 cases of head and neck squamous cell skin carcinoma. NOS2 expression was located in cells of the basal cell layer of epidermis, but was significantly increased in tumour epithelium. No correlations were found between NOS2 expression and tumour size or degree of tumour differentiation. The raised NO concentrations in these tumours may provide one explanation for their more aggressive behaviour than that of the slower-growing basal cell carcinoma.