HPV infection in squamous cell carcinomas arising from different mucosal sites of the head and neck region. Is p16 immunohistochemistry a reliable surrogate marker?

BACKGROUND: Human papillomavirus 16 infection has been proven to be associated with oropharyngeal squamous cell carcinomas (SCCs) and is probably the main reason of the reported increase in the incidence. The role of high-risk (HR) HPV for carcinogenesis of other sites in the head and neck awaits confirmation. With the aim to evaluate the prevalence of HPV infection and the reliability of different diagnostic tools in SCCs of different sites, 109 consecutive untreated head and neck SCCs were enrolled, and fresh tumour samples collected. METHODS: Human papillomavirus DNA was detected by Digene Hybrid Capture 2 (HC2). Human papillomavirus E6 and E7 mRNA were detected by NucliSENS EasyQ HPVv1. P16 expression was evaluated by immunohistochemistry. RESULTS: In all, 12.84% of cases were infected by HR genotypes and 1.84% by low-risk genotypes. Human papillomavirus 16 accounted for 87% of HR infections. The overall agreement between DNA and RNA detection is 99.1%. Although p16 expression clearly correlates with HPV infection (P=0.0051), the inter-rater agreement is poor (k=0.27). The oropharynx showed the highest HR HPV infection rate (47.6%) and was also the only site in which p16 immunohistochemistry revealed to be a fair, but not excellent, diagnostic assay (κ=0.61). CONCLUSION: The prognostic role of HR HPV infection in oropharyngeal oncology, with its potential clinical applications, underscores the need for a consensus on the most appropriate detection methods. The present results suggest that viral mRNA detection could be the standard for fresh samples, whereas DNA detection could be routinely used in formalin-fixed, paraffin-embedded samples.

Prevalence of Malnutrition in Older Hospitalized Cancer Patients: A Multicenter and Multiregional Study.

BACKGROUND: Malnutrition is frequent in older cancer patients, with a prevalence that ranges from 25% to 85%. The aging process is associated with several physiological changes, which may have implications for nutritional status. Screening tools can be useful for identifying malnutrition status among older patients with cancer. METHODS: A hospital-based multicenter cohort study that included 44 institutions in Brazil. The Mini Nutritional Assessment-Short Form (MNA-SF) was administered to 3061 older hospitalized cancer patients within 48 hoursof admission. The Kolmogorov-Smirnov test was used to test the sample distribution, considering sex, age range, calf circumference, body mass index, and MNA-SF score and classification. The categorical data were expressed by frequencies (n) and percentages (%)and compared using the chi-square test or Tukey test. RESULTS: According to the results of the MNA-SF, 33.4% of the patients were malnourished, 39.3% were at risk of malnutrition, and 27.3% were classified as having normal nutritional status. Length of hospital stay (in days) was found to be longer for those patients with a poorer nutritional status (malnourished: 7.07±7.58; at risk of malnutrition: 5.45±10.73; normal status: 3.9±5,84; p <0.001). CONCLUSIONS: The prevalence of malnutrition and nutritional risk is high in older hospitalized cancer patients in all the regions of Brazil and a worse nutritional status is associated with a longer hospital stay. Using a low-cost, effective nutritional screening tool for older cancer patients will enable specialized nutritional interventions and avoid inequities in the quality of cancer care worldwide.

Ruminant-like digestion of the langur monkey.

The adaptation of langur monkeys to a laboratory environment has made possible a detailed investigation of their digestive physiology. The diverticular form of the langur stomach permits a bacterial fermentation of the leafy diet, which results in important contributions to the nutrition of these primates. The demonstration of a ruminant-like digestion in langurs extends the known taxonomic distribution of this digestive adaptation.

Malnutrition associated with nutrition impact symptoms and localization of the disease: Results of a multicentric research on oncological nutrition.

BACKGROUND & AIMS: Malnutrition in cancer is an independent factor associated with negative clinical outcomes. The aim was to evaluate the prevalence and independent risk factors for malnutrition in hospitalized cancer patients using the Patient-Generated Subjective Global Assessment (PG-SGA). METHODS: We evaluated 4783 cancer patients, aged ≥20 years, in a hospital-based, multicenter, cross-sectional study. Patients were classified as well-nourished (PG-SGA Stage A), moderate/suspected malnutrition (PG-SGA Stage B), or severely malnourished (PG-SGA Stage C), and provided a score to define required nutritional interventions. Multivariate analysis was composed of the odds ratio (OR) estimated by ordinal polytomous logistic regression. RESULTS: 45.3% were classified as Stage B and 11.8% as Stage C. Moreover, 45.3% of the patients presented a need for nutritional intervention. The variables that presented the highest ORs for Stage B or Stage C were: problems with swallowing (OR 2.8, 95% confidence interval (CI) 2.2-3.4, p < 0.001), loss of appetite (OR 1.9, 95% CI 1.6-2.3, p < 0.001), vomiting (OR 1.8, 95% CI 1.5-2.3, p < 0.001), presence of more than 3 nutrition impact symptoms (OR 8.3, 95% CI 5.8-12, p < 0.001), and cancer site: lung (OR 4.6, 95% CI 3.2-6.6, p < 0.001), upper digestive cancer (OR 3.7, 95% CI 2.7-5.2, p < 0.001), and head and neck cancer (OR 3.7, 95% CI 2.7-5.2, p < 0.001). The score for Worksheet 4 on the PG-SGA had a higher association with malnutrition (OR 7.3, 95% CI 6.6-8.2, p < 0.001). CONCLUSIONS: Malnutrition is highly prevalent in cancer patients in Brazil, and is associated with nutritional impact symptoms, cancer site and age ≥65 years.

Autologous stem cell transplantation: sequential production of hematopoietic cytokines underlying granulocyte recovery.

We investigated the serum concentrations of a variety of cytokines [granulocyte-macrophage-colony-stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), interleukin (IL) 1 alpha, IL-3, IL-6, IL-8, erythropoietin, tumor necrosis factor alpha, gamma-interferon in 10 patients with advanced ovarian cancer undergoing autologous peripheral blood stem cell (PBSC) harvesting followed by treatment with high-dose cisplatin, etoposide, and carboplatin and PBSC transplantation (chemotherapy was administered on days 1 through 3, PBSCT on day 6). Preliminary observations on cytokine serum levels were performed for 4 patients; on this basis, the kinetics of cytokines was then investigated in greater detail at closely sequential times in 6 further patients. We observed a consistent pattern of sequential GM-CSF, G-CSF, and IL-8 release after chemotherapy/PBSCT in all 10 cases, including the 6 patients monitored in detail: (a) at days 5-10 a GM-CSF peak; (b) at days 12-14 a pronounced release of both G-CSF and IL-8, which always preceded granulocyte recovery by approximately 7 days. At days 17-23, a second GM-CSF peak was monitored in 5 of the 6 patients analyzed in detail, as well as in the other 4 cases. Particularly relevant are the observations that: (a) the peak of G-CSF serum concentration and neutrophil number in the recovery phase are strikingly and directly correlated, thus indicating a key role for G-CSF in granulocyte rescue; (b) the time courses of G-CSF and IL-8 levels are strictly parallel, thereby suggesting a coordinate stimulus for production of granulocytes, mediated by G-CSF, and their activation/migration capacity, mediated by IL-8. Results were essentially negative for IL-3, tumor necrosis factor alpha, and gamma-interferon concentrations (except in one case for each cytokine). An early peak of IL-1 alpha was observed in all 3 analyzed patients, while an IL-6 peak was monitored at days 13-15 in all 4 patients analyzed in detail. The present results indicate a sequential coordinate pattern of cytokine release after ablative therapy and PBSCT and shed light on the mechanisms mediating the recovery of granulocytes, and more generally of hematopoiesis, after stem cell transplantation. Furthermore, these studies may contribute to the design of improved protocols for cytokine administration following myelosuppressive anticancer therapy, as well as to the prediction of granulocytic response.

Interleukins 6 and 8 as mediators of acute phase response in acute myocardial infarction.

The present study provides evidence that interleukin (IL)-6 and IL-8 are the main endogenous mediators of acute phase response in patients with myocardial infarction. This conclusion was supported by the observation of a strict relation between IL-6 elevation and the extent of myocardial tissue damage and rise in body temperature.

Blood levels of erythropoietin in congestive heart failure and correlation with clinical, hemodynamic, and hormonal profiles.

Plasma levels of erythropoietin (mU/ml) were measured in patients with congestive heart failure (CHF) (n = 108) and in a control group of normal subjects (n = 45). In normal subjects, plasma levels of erythropoietin were 1.9 +/- 0.2. In patients with CHF, plasma levels of erythropoietin increased progressively according to New York Heart Association (NYHA) class (I: 1.4 +/- 0.2, n = 28; II: 5.4 +/- 0.8, n = 27; III: 9.6 +/- 2, n = 32; IV: 34 +/- 8, n = 21; F = 57.7, p < 0.001) and were significantly higher in NYHA classes II, III, and IV than in normal subjects. Plasma erythropoietin significantly decreased (from 43 +/- 14 to 12 +/- 3 mU/ml, p < 0.01) in patients with severe CHF (n = 9) when enalapril (20 mg/day administered orally) was added to long-term treatment for 3 weeks. Finally, in a subgroup of patients with NYHA class IV CHF (n = 9) and high plasma erythropoietin levels (37 +/- 9 mU/ml), packed red blood cell volume, assessed by the iodine-125-albumin dilution method, was higher than that in normal subjects (n = 11) (2,616 +/- 235 vs 2,028 +/- 119 ml, p < 0.05). The present study demonstrates that plasma erythropoietin levels are elevated in a large cohort of patients with CHF of varying etiology, and that this increase is related to the progression of the disease. The increase in circulating erythropoietin is associated with augmented packed red blood cell volume in patients with severe CHF. These results suggest a participation of erythropoietin in the complex neurohormonal response that occurs in CHF.

Inhibition of lymphocyte blastogenic response in healthy donors treated with recombinant human granulocyte colony-stimulating factor (rhG-CSF): possible role of lactoferrin and interleukin-1 receptor antagonist.

The effects of rhG-CSF on lymphocyte blastogenesis were evaluated in six healthy donors, submitted to progenitor cell mobilization for allogeneic transplantation. Neutrophil, monocyte and lymphocyte count increased 6.7-fold, 5.3-fold and 2.0-fold on day +4 of rhG-CSF as compared with baseline. The DNA stimulation index (DNA SI) of 72 h phytohemagglutinin (PHA)-treated cultures decreased from 20% (15-35.5) prior to rhG-CSF to 6.7% (1.5-11.9; P = 0.0026), 8% (4-12; P = 0.0091) and 15% (9-22; P = 0.0091) on days +2, +4 and +6; similarly, reactivity to concanavalin A decreased from 18% (12-20) to 1.8% (0.5-7; P < 0.01), 3% (2-8; P < 0.01) and 5% (2-11; P = 0.009). No changes of lymphocyte response to pokeweed mitogen were observed. DNA SI of PHA-treated cultures inversely correlated with neutrophil and monocyte count. IL-1 receptor antagonist (IL-1ra) and lactoferrin (Lf) plasma levels sharply increased and correlated with neutrophil and monocyte count. IL-10 increased five-fold on day +2, returned to pretreatment values thereafter and did not show any correlation with DNA SI, suggesting that it was not responsible for the observed phenomena. Interestingly, DNA SI of PHA-treated cultures inversely correlated with IL-1ra and Lf levels. CD3+ and CD19+ lymphocyte activation status, ie CD23, CD25, CD30 and HLA-DR coexpression, was not affected by rhG-CSF administration. Pharmacological doses of rhG-CSF in healthy donors inhibit lymphocyte blastogenesis via an increased production and/or release of immunoregulatory soluble mediators, ie IL-1ra and Lf, by primed neutrophils and monocytes.

A comparison of alphaprodine and meperidine pharmacokinetics.

We have developed a gas-liquid chromatographic analysis for measuring plasma alphaprodine. The analysis is sufficiently sensitive for studying therapeutic-dose pharmacokinetics in man. Single intravenous bolus injections of either alphaprodine or meperidine were given to volunteer subjects. Plasma concentration data were fitted to a biexpoential equation and pharmacokinetic parameters were calculated. Consistent with alphaprodine's shorter clinical duration compared to meperidine, we found that plasma levels of alphaprodine decline at a more rapid rate. Plasma clearances for the two narcotics are nearly identical (1.04 l./kg/min for alphaprodine and 1.01 l./kg/min for meperidine). The apparent volume of distribution for alphaprodine is smaller than for meperidine (1.90 l./kg for alphaprodine and 3.37 l./kg for meperidine).

Nicotine- or epinephrine-induced uteroplacental vasoconstriction and fetal growth in the rat.

We examined the relationship between nicotine-induced vasoconstriction in pregnant rat dams and fetal growth during the third trimester of pregnancy. Pregnant rats were continuously treated between days 13 and 19 of gestation with either nicotine (9.6, 4.8 or 2.4 mg/kg/day), epinephrine (0.72 microgram/kg/day), or saline via continuous infusion from a subcutaneously implanted osmotic minipump. Placental weights in rats treated with high dose nicotine and dams' body weights were severely reduced. However, fetal weights were not affected. Blood flows in uterus and placenta were quantified by measurement of tissue content of 85Sr-labelled microspheres injected via a carotid artery catheter. Both nicotine and epinephrine caused a significant reduction (> 40%) in uterine and placental blood flow. We conclude that vasoconstriction alone as a result of nicotine or epinephrine administration during the last trimester of gestation does not necessarily reduce nutrient supply to the fetus and does not affect fetal growth in rats.

Blood pressure response to phenylephrine infusion in halothane-anesthetized dogs given acetylpromazine maleate.

To quantitate acetylpromazine-induced alpha-adrenergic receptor blockade, phenylephrine was infused into dogs. The amount of phenylephrine necessary to increase the mean arterial blood pressure (MAP) 50% above base line, with or without the prior administration of acetylpromazine, served to quantify the degree of acetylpromazine-induced alpha-adrenergic receptor blockade. Seven dogs were anesthetized with thiopental, maintained on halothane in oxygen, and mechanically ventilated. All infusions were made through a catheter in the cephalic vein. Continuous recordings were made of MAP and a lead II ECG. After induction of anesthesia, instrumentation, and stabilization of heart rate, MAP, and ventilation, 6 group I dogs were infused with phenylephrine until a 50% increase in MAP was recorded (phenylephrine control). On subsequent research days, each dog was anesthetized, instrumented as described, and given (IV) 1 of 3 dosages of acetylpromazine in the following order--0.05, 0.125, and 0.25 mg/kg. The dose of phenylephrine necessary to increase MAP 50% in the presence of acetylpromazine was recorded. Five group II dogs were studied as in group I, but each dog was given (IM) atropine (0.04 mg/kg) before anesthetization. Two dosages of acetylpromazine were studied in the following order--0.05 and 0.25 mg/kg. Group I dogs, when compared with their phenylephrine controls, were given significantly more phenylephrine to raise MAP 50% at each dose of acetylpromazine studied. The same trend was observed in group II dogs, but at smaller doses of phenylephrine, probably as a result of the positive chronotropic effect of atropine on the heart.

Blood gas and catecholamine levels in capture stressed desert bighorn sheep.

Forty-seven bighorn sheep (Ovis canadensis nelsoni) were captured within a 3-day period in December, 1989 as part of a California Department of Fish and Game effort to repopulate historic ranges in California. They were captured on the Mojave Desert in the Kelso Mountains near Old Dad Peak, San Bernardino County, California. Venous blood gases measured at the site of capture demonstrated a severe metabolic acidosis (base deficit, 23 mEq/liter), with no evidence of respiratory acidosis. There were moderately elevated plasma epinephrine (1.25 ng/ml), norepinephrine (2.60 ng/ml), and dopamine (114 pg/ml) levels. These data appear to reflect animals that have been moderately stressed. These acid-base-catecholamine values differ from values in resting domestic sheep, and are similar to those reported in greyhounds after brief strenuous exercise.

Congenital stricture of male urethra.

Seven cases of anterior urethral stricture were studied. The diagnosis was made by exclusion of the inflammatory and traumatic causes. In three cases the stricture was located in the bulbar and in four cases in the penile urethra. The results obtained were good, with a follow-up of 5 to 50 months. Considerations are made about embryogenic, diagnostic, clinical picture and treatment. The verification of extensive lesions of the urethra determined the use of more complex surgical techniques of Johansen's urethroplasty devised for the treatment of this kind of pathology.

Evaluation of the impairment of spermatogenesis in varicocele through semen analysis and testicular biopsy.

Twenty-four patients with varicocele and 26 normal men were analysed according to the seminal ejaculate and testicular biopsy. Patients were divided into two groups based on the criteria of sperm normality and abnormality, respectively. Hypospermatogenesis with both normal and abnormal seminal fluid occurred in the varicocele patients, but no statistical differences could be established when compared with each other. We concluded that testicular biopsy is not useful in the evaluation of patients with varicocele.

Mice heterozygous for the oxytocin receptor gene (Oxtr(+/-)) show impaired social behaviour but not increased aggression or cognitive inflexibility: evidence of a selective haploinsufficiency gene effect.

We characterised the behavioural phenotype of mice heterozygous (Oxtr(+/-)) for the oxytocin receptor gene (Oxtr) and compared it with that of Oxtr null mice (Oxtr(-/-)), which display autistic-like behaviours, including impaired sociability and preference for social novelty, impaired cognitive flexibility, and increased aggression. Similar to Oxtr(-/-) mice, the Oxtr(+/-) showed impaired sociability and preference for social novelty but, unlike the null genotype, their cognitive flexibility and aggression were normal. By autoradiography, Oxtr(+/-) mice were found to have approximately 50% fewer oxytocin receptors (OXTRs) in all of the examined brain regions. Thus, because a partial reduction in Oxtr gene expression is sufficient to compromise social behaviour, the Oxtr acts as a haploinsufficient gene. Furthermore, the inactivation of the Oxtr gene affects specific behaviours in a dose-dependent manner: social behaviour is sensitive to even a partial reduction in Oxtr gene expression, whereas defects in aggression and cognitive flexibility require the complete inactivation of the Oxtr gene to emerge. We then investigated the rescue of the Oxtr(+/-) social deficits by oxytocin (OT) and Thr(4)Gly(7)OT (TGOT) administered i.c.v. at different doses. TGOT was more potent than OT in rescuing sociability and social novelty in both genotypes. Furthermore, the TGOT doses that reverted impaired sociability and preference for social novelty in Oxtr(+/-) were lower than those required in Oxtr(-/-), thus suggesting that the rescue effect is mediated by OXTR in Oxtr(+/-) and by other receptors (presumably vasopressin V1a receptors) in Oxtr(-/-). In line with this, a low dose of the selective oxytocin antagonist desGlyDTyrOVT blocks the rescue effect of TGOT only in the Oxtr(+/-) genotype, whereas the less selective antagonist SR49059 blocks rescue in both genotypes. In conclusion, the Oxtr(+/-) mouse is a unique animal model for investigating how partial loss of the Oxtr gene impair social interactions, and for designing pharmacological rescue strategies.

A computer evaluation of the ratio of the diastolic pressure-time index to the time-tension index from three arterial sites in dogs.

The ratio of the area under the diastolic portion of the arterial pulse pressure trace (diastolic pressure-time index; DPTI) to the area under the systolic component of the arterial pulse pressure trace (time-tension index; TTI) has been used to predict the relationship of myocardial blood supply to oxygen demand. Since introduction of the DPTI-to-TTI ratio as a measurement of this relationship, the accepted critical DPTI:TTI value below which subendocardial ischemia may occur has decreased by almost 50%. This lower critical value has come about as more clinical experience has been gained, particularly in patients with an arterial catheter in the arm. To investigate a potential cause for this decrease, we studied a canine model by pulse transduction from the central aorta (the site traditionally used for these ratio determinations), as well as the femoral artery and the median forepaw artery. Following inotropic and ventricular loading interventions, the changes in the DPTI:TTI, calculated by a special-purpose computer, were exaggerated by approximately 25% in the peripheral arterial measurements. The peripheral arterial sites had consistently higher systolic waveforms and consistently lower, broader diastolic waveforms than those for the central aortic site. This type of configurational change is probably a major cause of the differences among ratios from the three sites in our study, and it helps to explain why the value of the critical ratio has been a subject of controversy.

Determinants and genesis of canine pneumocardiogram.

The pneumocardiogram measures the to-and-fro movement of gas from the lung with each heartbeat during apnea. Six mongrel dogs were prepared with chronic ascending aortic or pulmonary artery flow probes, corresponding occlusion cuffs and intraventricular pressure transducers. Under halothane anesthesia, the flow pneumocardiogram (PnCG) and its time derivative (acceleration pneumocardiogram or dPn/dt) were transduced during apnea by a small high-gain pneumotachograph. All variables were recorded on paper. The effects on the pneumocardiogram of great vessel occlusion, changes in cardiac loading, small airway patency, and chest wall integrity were investigated. Four of the animals were treated subsequently to produce a chemical cardiac denervation and restudied. The systolic component of the dPn/dt which correlates with myocardial contractility (IJ wave) was dependent primarily on left ventricular (LV) ejection, markedly affected by cardiac preload and modified by afterload changes to a lesser degree. Small airway closure obliterated the wave form, while opening the chest wall attenuated the dPn/dt IJ wave by 25%. This study shows the dPn/dt IJ wave is anatomically related to LV performance and responds to changes in LV function similar to other noninvasive cardiac measurements.

Determination of halothane ED-50 in Mongolian gerbils (Meriones unguiculatus).

The effective dose-50 (ED-50) for halothane anesthesia was studied in 45 male Mongolian gerbils at age 12 weeks. Close control of circadian variation and animal temperature was maintained. Regression analysis for the percent of animals moving in response to a standard stimulus revealed an ED-50 of 1.32 volumes %, a concentration that approximates the ED-50 of the rat.

Cardiovascular effects of atropine sulfate preparations in vagotomized dogs under halothane anesthesia.

The cardiovascular effects of 3 preparations of atropine sulfate were studied acutely in open-chest, vagotomized dogs under endotracheal halothane anesthesia. Indices of myocardial performance (LVdp/dt/CPIP and maximum ascending aortic blood acceleration) showed insignificant changes when varying doses of IV atropine (0.04 mg/kg and 0.04 mg/kg) were given. However, mean ascending aortic pressure fell by 20 percent following the larger doses of 2 commercial preparations containing antibacterial preservatives, and only 9 percent following a "pure" (USP) atropine preparation. Calculated changes in systemic vascular resistance closely followed actual pressure values. These results indicate that atropine, even in large doses, causes little or no depression of ventricular function independently of its chronotropic action. However, atropine does cause a fall in blood pressure, seemingly due to peripheral vasodilation, particularly in commercial preparations containing preservatives.