On the mechanism of solid-state phase transitions in molecular crystals - the role of cooperative motion in (quasi)racemic linear amino acids.

During single-crystal-to-single-crystal (SCSC) phase transitions, a polymorph of a compound can transform to a more stable form while remaining in the solid state. By understanding the mechanism of these transitions, strategies can be developed to control this phenomenon. This is particularly important in the pharmaceutical industry, but also relevant for other industries such as the food and agrochemical industries. Although extensive literature exists on SCSC phase transitions in inorganic crystals, it is unclear whether their classications and mechanisms translate to molecular crystals, with weaker interactions and more steric hindrance. A comparitive study of SCSC phase transitions in aliphatic linear-chain amino acid crystals, both racemates and quasi-racemates, is presented. A total of 34 transitions are considered and most are classified according to their structural change during the transition. Transitions without torsional changes show very different characteristics, such as transition temperature, enthalpy and free energy, compared with transitions that involve torsional changes. These differences can be rationalized using classical nucleation theory and in terms of a difference in mechanism; torsional changes occur in a molecule-by-molecule fashion, whereas transitions without torsional changes involve cooperative motion with multiple molecules at the same time.

Carbon dating cancer: defining the chronology of metastatic progression in colorectal cancer.

Background: Patients often ask oncologists how long a cancer has been present before causing symptoms or spreading to other organs. The evolutionary trajectory of cancers can be defined using phylogenetic approaches but lack of chronological references makes dating the exact onset of tumours very challenging. Patients and methods: Here, we describe the case of a colorectal cancer (CRC) patient presenting with synchronous lung metastasis and metachronous thyroid, chest wall and urinary tract metastases over the course of 5 years. The chest wall metastasis was caused by needle tract seeding, implying a known time of onset. Using whole genome sequencing data from primary and metastatic sites we inferred the complete chronology of the cancer by exploiting the time of needle tract seeding as an in vivo 'stopwatch'. This approach allowed us to follow the progression of the disease back in time, dating each ancestral node of the phylogenetic tree in the past history of the tumour. We used a Bayesian phylogenomic approach, which accounts for possible dynamic changes in mutational rate, to reconstruct the phylogenetic tree and effectively 'carbon date' the malignant progression. Results: The primary colon cancer emerged between 5 and 8 years before the clinical diagnosis. The primary tumour metastasized to the lung and the thyroid within a year from its onset. The thyroid lesion presented as a tumour-to-tumour deposit within a benign Hurthle adenoma. Despite rapid metastatic progression from the primary tumour, the patient showed an indolent disease course. Primary cancer and metastases were microsatellite stable and displayed low chromosomal instability. Neo-antigen analysis suggested minimal immunogenicity. Conclusion: Our data provide the first in vivo experimental evidence documenting the timing of metastatic progression in CRC and suggest that genomic instability might be more important than the metastatic potential of the primary cancer in dictating CRC fate.

Melanoma.

The incidence of malignant melanoma in the UK is still rising despite public health warnings about the risks of excessive sun exposure. This aggressive tumour can metastasize to virtually any organ, even years after resection of the primary lesion and cause a variety of radiological appearances. This review provides examples of both typical and non-specific imaging features of melanoma metastases, as well as examples of primary choroidal melanoma.

Altered cardiac collagen and associated changes in diastolic function of infarcted rat hearts.

OBJECTIVE: Anti-inflammatory drugs have been shown to modulate collagen deposition during myocardial infarction (MI) induced remodeling. Chronic effects of methylprednisolone (5 mg/kg/day) and low-dose aspirin (25 mg/kg/day) on cardiac collagen and left ventricular diastolic function were studied in rat hearts, 21 days after MI. METHODS: Left ventricular function was assessed at baseline and after beta-adrenergic stimulation with isoproterenol in isolated perfused hearts, using an intraventricular balloon. After diastolic arrest, left ventricular pressure-volume curves were obtained. Left ventricular dilation was defined as the corresponding left ventricular volume at 20 mmHg left ventricular diastolic pressure. In histological sections, perivascular and interstitial collagen content were quantified morphometrically as the Sirius Red positive area in the non-infarcted interventricular septum. RESULTS: Impaired baseline left ventricular function of MI-hearts was improved by methylprednisolone but not by low-dose aspirin. Isoprotenerol significantly enhanced systolic function in all hearts, whereas it augmented the decrease in left ventricular diastolic pressure only in methylprednisolone-treated MI-hearts. The rightward shift of the pressure-volume curve after MI was aggravated by methylprednisolone but not with low-dose aspirin treatment. Low-dose aspirin reduced perivascular but not interstitial collagen whereas methylprednisolone decreased both perivascular and interstitial collagen. CONCLUSIONS: Our findings indicate that MI-induced collagen deposition in the spared myocardium can be affected by chronic therapy with low-dose aspirin or methylprednisolone. The effects on interstitial collagen seemed reflected in an altered left ventricular diastolic function.

Early captopril prevents myocardial infarction-induced hypertrophy but not angiogenesis.

Delayed captopril, started after the healing phase of myocardial infarction, improves perfusion by reducing tissue weight without affecting the vascular capacity of the heart. Early captopril, during the healing phase, prevents reactive hypertrophy, but the effects on angiogenesis are unknown. Therefore, the effects of early captopril (2 g/l drinking water, from 1 day until 3 weeks after myocardial infarction) on regional coronary flow related to tissue mass, were studied in isolated perfused hearts from rats, subjected to coronary artery ligation. Regional maximal vascular capacity was measured during nitroprusside-induced vasodilation, using radioactive microspheres. Maximal vascular capacity was not changed by captopril. Reactive hypertrophy in infarcted hearts only reached statistical significance in the left ventricular free wall. Since captopril prevented hypertrophy but did not affect regional capacity, peak tissue perfusion was improved. Indicating effects on metabolism, captopril restored the increased lactate/purine ratio in infarcted hearts. Thus, early captopril treatment prevented post-myocardial infarction hypertrophy but did not suppress angiogenesis, thus beneficially influencing the vascularization/tissue mass ratio, probably reflected by preservation of aerobic metabolism.

Low-dose aspirin improves in vivo hemodynamics in conscious, chronically infarcted rats.

OBJECTIVE: The equivalent of the clinically used low (= antiplatelet)-dose aspirin, inhibited collagen deposition in the non-infarcted myocardium in rats with myocardial infarction. In the present study, the in vivo hemodynamic consequences of this daily low-dose aspirin were investigated in conscious, chronically instrumented, infarcted rats. METHODS: Rats, treated with 25 mg/kg aspirin daily from 2 days before to 3 weeks after coronary artery ligation, were chronically instrumented with an electromagnetic flow-probe and arterial and venous catheters, to record cardiac output, and arterial and venous blood pressure, respectively, in the conscious freely moving animal. In parallel, isolated hearts were studied with regard to left ventricular stiffness (pressure/volume relationships), maximal cardiac perfusion (adenosine), and in vitro heart rate and beta-adrenergic responsiveness. Plasma catecholamine levels were measured. RESULTS: Aspirin normalized the increased heart rate after infarction, at a preserved cardiac output. This was accompanied by a (non-significant) increase in stroke volume, at unchanged cardiac loading conditions. The lower heart rate after aspirin was due to reduced intrinsic heart rate rather than to lower sympathetic activation of the heart, since similar effects were observed in isolated perfused hearts, while circulating levels of catecholamines and beta-adrenergic responsiveness were not influenced. The improved stroke volume was not explained by reduced left ventricular stiffness or increased maximal perfusion after aspirin. CONCLUSION: In addition to the antithrombotic action, effects of low-dose aspirin on cardiac remodeling could be associated with favorable hemodynamic effects, as reflected by a lower heart rate for the same cardiac output. Although the underlying mechanisms are still unknown, it suggests a clinically relevant beneficial effect which deserves further investigation.

Regionally different vascular response to vasoactive substances in the remodelled infarcted rat heart; aberrant vasculature in the infarct scar.

Remodelling after myocardial infarction (MI) is associated with vascular adaption, increasing vascular capacity of non-infarcted myocardium, and angiogenesis in the infarcted part during wound healing and scarring. We investigated regional vascular reactivity in the infarcted rat heart. Transmural infarction of the left ventricular free wall was induced by coronary artery ligation. After 3 weeks, regional flow during maximal vasodilation (nitroprusside, NPR) and submaximal vasoconstriction (arginine-vasopressin, AVP) were studied in buffer-perfused hearts. The main findings were: (1) a reduced vasodilator response (NPR) in the viable part of the left ventricular free wall, where hypertrophy was most pronounced, resulting in reduced maximal tissue perfusion of the myocardium bordering the scar (19.7 + 0.6 v 25.7 + 1.2 ml/min.g), whereas perfusion of other non-infarcted regions was preserved. (2) A 54% lower vasodilator response (NPR) and a 25% stronger vasoconstriction (AVP) in scar tissue compared to viable parts of MI hearts. Microscopy showed thicker walls of resistance arteries in scar tissue than in viable parts of MI hearts or in sham hearts, morphometrically substantiated by two- to three-fold greater wall/lumen ratios. These data indicate a deviant response of scar vessels of MI hearts, and in the non-infarcted part, a reduced coronary reserve in the most hypertrophied region. Whereas the former may be caused by different vessel structure, the reduced vasodilator reserve of the spared part of the left ventricular free wall may indicate vasodilation at rest due to insufficient vascular growth. Thus, the most hypertrophied region would be at the highest risk of further ischemic damage.

Determinants of coronary reserve in rats subjected to coronary artery ligation or aortic banding.

OBJECTIVE: We investigated whether decreased coronary reserve in hearts after coronary artery ligation or in hearts from rats after aortic banding can be related to remodeling of resistance arteries. METHODS: Maximal coronary flow (absolute flow) and cardiac perfusion (flow corrected for heart weight) were determined in isolated, perfused rat hearts after adenosine or nitroprusside, at 3 and 8 weeks after coronary artery ligation or 4-5 weeks after aortic banding. Perivascular collagen and medial thickness of resistance arteries were determined by morphometry. RESULTS: maximal coronary flow of infarcted hearts had been restored to sham values at 3 weeks. Growth of cardiac muscle mass from 3 to 8 weeks exceeded the increase in maximal coronary flow, leading to a decreased perfusion at 8 weeks. A slight, transient increase in perivascular collagen, but no medial hypertrophy, was found after infarction. After aortic banding perivascular fibrosis and medial hypertrophy led to a decreased maximal coronary flow in both the hypertrophied left and the non-hypertrophied right ventricle. Consequently, perfusion of the left ventricle was most severely reduced. CONCLUSIONS: Reduced maximal perfusion after aortic banding is determined by both cardiac hypertrophy and vascular remodeling. In contrast, during infarction-induced remodeling, reduction of perfusion is not determined by vascular remodeling, but mainly by disproportional cardiac hypertrophy relative to vascular growth.

Sensitivity to ischemia of chronically infarcted rat hearts; effects of long-term captopril treatment.

Myocardial infarction induced hypertrophy of non-infarcted myocardium, in parallel with interstitial and perivascular fibrosis and a decreased capillary density, could increase sensitivity to ischemia. The structural cardiac changes can be reversed by long-term captopril treatment. In the present study, ischemic sensitivity in relation to cardiac perfusion was studied in isolated, perfused hearts of untreated and captopril-treated infarcted rats. In chronically (8 weeks) infarcted hearts, maximal vasodilation in response to administered adenosine and nitroprusside, as well as to endogenously released vasodilators during reperfusion, was decreased, suggesting impaired cardiac perfusion. Ischemic release of purines and lactate was reduced in infarcted hearts, indicating decreased sensitivity to ischemia of the remodeled myocardium. Captopril treatment (3-8 weeks post myocardial infarction), which reversed hypertrophy without affecting the flow capacity of the coronary vascular bed, restored maximal cardiac perfusion. Ischemic ATP breakdown was not affected by captopril, whereas lactate release was even further reduced, suggesting alterations towards a more aerobic ATP production. These data indicate that despite the reduced maximal cardiac perfusion, the remodeled myocardium of infarcted hearts is less sensitive to ischemia. Reversal of hypertrophy by chronic captopril restored maximal cardiac perfusion and led to a better preservation of aerobic ATP production during ischemia.

'Quality of life' after therapy in rats with myocardial infarction: dissociation between hemodynamic and behavioral improvement.

Rats with myocardial infarction provide a clinically relevant model for hemodynamic and survival studies. Moreover, behavioral changes in this model, i.e. increased anxiety and reduced interest in environment and social interactions, mimic aspects of the reduced quality of life of patients. In the present study, we investigated whether pharmacological treatment that is known to improve hemodynamics and prognosis could also affect the behavioral changes associated with quality of life. Rats with 3-week-old infarcts were treated with intermittent dobutamine (1 mg/kg i.p., twice daily) or captopril (2 milligrams in drinking water). After 2 weeks of treatment, when from previous studies hemodynamics are expected to be restored, behavioral tests were performed. In the free exploration test, which primarily evaluates exploratory behavior, dobutamine normalized the reduced interest in the environment. In the standard open field and social interaction tests, which also include an anxiety component, the beneficial effects of dobutamine were not observed. In contrast, captopril normalized all behavioral changes that indicated increased anxiety. In conclusion, the expected similar hemodynamic improvement with dobutamine and captopril treatment resulted in improvement of different aspects of the changed behavior of rats with myocardial infarction, indicating that there is no direct relationship between hemodynamics and quality of life. The behavioral tests used, in combination with our previously described functional hemodynamic measurements, could provide a new basis for evaluating the effects of therapy on hemodynamic function as well as the quality of life.

Chronic aspirin treatment affects collagen deposition in non-infarcted myocardium during remodeling after coronary artery ligation in the rat.

Low-dose aspirin (acetylsalicylic acid; ASA), inhibiting platelet thromboxane production in favor of endothelium formation of prostaglandins, is successfully used as primary or secondary prophylaxis against myocardial infarction. Although prognosis may be improved, effects of long-term ASA treatment on wound healing and cardiac remodeling are not well understood. The aim of the present study was to mimic the clinical situation by inducing myocardial infarction in low-dose ASA (25 mg/kg/day, i.p.) pretreated rats, and to determine effects on plasma eicosanoid levels, cardiac hypertrophy and collagen deposition, and left ventricular function during continued ASA treatment. The effects of this dose were verified to selectively inhibit platelet thromboxane production, and lower plasma levels of thromboxane, but did not affect plasma levels of prostacyclin and prostaglandin E2 during the acute inflammatory stage following myocardial infarction. As measured by heart dry weight/body weight, cardiac hypertrophy was not affected by ASA treatment. However, interstitial fibrosis in the spared myocardium as well as perivascular fibrosis, associated with infarction-induced cardiac remodeling, were affected by ASA treatment. Replacement fibrosis in the infarct itself, considered as representing wound healing, was not significantly influenced by ASA treatment. Wall thinning following infarction was not aggravated, nor did treatment influence left ventricular cavity diameter in a relaxed state. Results from in vitro left ventricular function measurements showed no effects on left ventricular peak velocity of contraction or relaxation after ASA treatment. In conclusion, although low-dose ASA may not be expected to have anti-inflammatory action, it did influence post-infarct cardiac remodeling by affecting interstitial and perivascular fibrosis. ASA treatment did not have effects on in vitro left ventricular dysfunction.

Stenting of the ureterovesical anastomosis in pediatric renal transplantation.

From January 1985 to July 1989, 36 children received a renal transplant at our hospital. Their ages ranged from 2 to 18 years. All patients had a standard neoureterocystostomy according to Lich-Grégoir. In the first 18 patients, no transanastomotic stent was placed. In the latter 18, a transanastomotic stent with or without suprapubic bladder drainage was performed. In the nonstented group, six severe urological complications occurred, two of which eventually resulted in loss of the transplant. In the stented group, only one severe urological complication occurred. No kidneys in this group were lost due to urological complications. The number of urinary tract infections in the nonstented group was the same as in the stented group.