Role of Matrix Degradation, Oxidative Stress, Inflammation & Trace Elements in COVID-19 Patients: A Multivariate Study from India.

The interrelationship between matrix degradation, oxidative stress, inflammation and trace elements can be speculated in COVID-19. The objective of the study was to evaluate the oxidative stress, inflammation and matrix degradation markers and trace elements in COVID-19 positive patients. A group of confirmed severe COVID-19 positive patients (n = 30) along with COVID-19 negative patients (n = 30) with similar symptoms were included. Both group of patients were assessed for oxidative stress markers, inflammatory cytokines, matrix metalloproteinase (MMP)s and their inhibitors along with trace elements in blood. All the data were subjected to univariate as well as multivariate analysis including PCA, PLS-DA, OPLS-DA. Diagnostic accuracy was tested by ROC curve analysis. Further relationship with Neutrophil/ lymphocyte (N/L) ratio was established if any. Increased oxidative stress, inflammation and matrix degradation is evidenced by significant rise in oxidative markers, inflammatory cytokines and MMP9/TIMP-1 ratio. Decreased Cu/Zn ratio is also observed in COVID-19 positive patients. Multivariate analysis identified SOD, Cu/Zn ratio, IL-6 and TOS, as effective discriminant among the two groups of patients. Further, accuracy was confirmed by ROC curves. Neutrophil/ lymphocyte (N/L) ratio, shows significant negative association with SOD (r= -0.75, p < 0.005) and Cu/Zn ratio (r = -0.88, p < 0.005). These data suggest the attributes of these biomarkers in disease severity. The potential use of these blood-based laboratory markers in disease prognosis seems promising and warrants further attention. Given by the symptoms and severity of the disease, it will be promising to monitor Cu/Zn ratio along with other prognostic indicators.

Study of angiogenesis in invasive breast carcinoma by morphometry and immunohistochemistry.

Background: Breast cancer is the leading cause of cancer-related deaths in Asia and is emerging as the commonest female malignancy. Angiogenesis or neovascularization is important for the growth and spread of malignant tumors, and quantitative assessment of angiogenesis may prove valuable in prognostication. This study was undertaken to quantify and explore angiogenesis with immunohistochemistry with CD 34, CD 105, and vascular endothelial growth factor (VEGF), as well as morphometric analysis and correlate with the grades of the invasive breast carcinoma. Methods: Angiogenesis was assessed by morphometry and immunohistochemistry. Seventy cases of invasive ductal carcinoma (IDC) and twenty-five benign cases as controls were included in the study. Morphometry was performed on the CD34 and CD105 (Endoglin) stained representative histologic sections with the use of a computerized digital photomicrograph system using image analyzing software. Morphometric analysis and evaluation of vascular parameters, i.e. microvessel density (MVD), microvessel caliber (VC), and total microvessel boundary density (TVBD), were calculated. Semiquantitative assessment of angiogenesis of VEGF-stained sections was done by scoring. Immunohistochemical staining was correlated with the histological grade of the tumors. MVD, mean VC, TVBD with their mean values, SD, and range were calculated using Statistical Package for The Social Sciences (Version 20). One-way analysis of variance (ANOVA) with Tukey HSD was performed to assess the difference of the parameters for the groups. Spearman rank correlation coefficients ρ were calculated. Results: The vascular parameters were significantly more in malignant lesions as compared to benign lesions and showed differences with increasing grade. Grades of breast carcinoma showed a mild positive correlation with VEGF (ρ = 0.467), MVD-CD34 (ρ = 0.422) and VC-CD34 (ρ = 0.482); and moderate positive correlation with TVBD-CD34 (ρ = 0.615), VC-CD105 (ρ = 0.527), and TVBD-CD105 (ρ = 0.354). When these parameters were compared with each other for all four groups, VEGF showed a mild positive correlation with MVD-CD34 (ρ = 0.295), TVBD-CD34 (ρ = 0.339), and TVBD-CD105 ((ρ = 0.277). MVD-CD105 showed a mild positive correlation with MVD-CD34 TVBD-CD105 also showed a strong positive correlation with MVD-CD34. VC-CD105 showed a moderate positive correlation with VC-CD34. CD 105 stained fewer but larger caliber vessels. Conclusions: In this study, vascular parameters showed significant differences in three grades of IDC with CD34. Differences were seen in vascular parameters stained with CD105 in three grades of IDC. Expression of VEGF also showed significant differences with positive correlations in the three grades of IDC. CD34 highlighted both old and newly formed microvessels. CD 105 stained fewer but larger caliber microvessels. VC-CD105 can be an extremely useful adjunct along with VEGF and CD34 to study angiogenesis of vessels in IDC.

An atypical presentation of toxic epidermal necrolysis without mucosal involvement.

Toxic epidermal necrolysis (TEN) is a severe adverse cutaneous drug reaction with ubiquitous involvement of mucosa. Drugs are identified as the main etiology in most cases. Cutaneous involvement in TEN occurs in the form of widespread painful erythematous macules, targetoid lesions, full-thickness or focal epidermal necrosis, whereas mucosal involvement involves oral, genital, and ocular mucous membranes along with preceding prodromal flu-like symptoms. Atypical presentations include involvement of only mucosa without involvement of skin. We report a rare case of TEN without any mucosal involvement.

Concordance of measures of left-ventricular hypertrophy in pediatric hypertension.

The American Academy of Pediatrics (AAP) recommends that any child diagnosed with hypertension have an echocardiogram to evaluate for the presence of left-ventricular (LV) hypertrophy (LVH) and advocates that LVH is an indication to initiate or intensify antihypertensive therapy. However, there is no consensus on the ideal method of defining LVH in the pediatric population. Many pediatric cardiologists rely on wall-thickness z-score of the LV posterior wall and/or interventricular septum to determine LVH. Yet, the AAP advocates using LV mass indexed to 2.7 (LVMI(2.7)) ≥ 51 g/m(2.7) to diagnose LVH. Recently, age-specific reference values for LVMI ≥ 95% were developed. The objective of the study was to determine the concordance between diagnosis of LVH by wall-thickness z-score and diagnosis by LVMI(2.7) criteria. A retrospective chart review was performed for subjects diagnosed with hypertension at a single tertiary care center (2009-2012). Echocardiogram reports were reviewed, and assessment of LVH was recorded. Diagnosis of LVH was assigned to each report reviewed according to three criteria: (1) LV wall-thickness z-score > 2.00; (2) age-specific reference values for LVMI(2.7) > 95th percentile; and (3) LVMI(2.7) > 51 g/m(2.7). Cohen's kappa statistic was used as a measurement of agreement between diagnosis by wall-thickness z-score and diagnosis using LVMI(2.7). A total of 159 echocardiograms in 109 subjects were reviewed. Subjects included 31 females and 77 males, age 13.2 ± 4.4 years, and 39 (42%) with a diagnosis of secondary hypertension. LVH was diagnosed in 31 cases (20%) based on increased wall-thickness z-score. Using LVMI(2.7) > 95%, LVH was found in 75 (47%) cases (mean LVMI(2.7)42.3 ± 17.2 g/m(2.7) [range 11.0-111 g/m(2.7)]). The wall-thickness z-score method agreed with LVMI(2.7) > 95% diagnosis 71% of the time (kappa 0.4). Using LVH criteria of LVMI(2.7) ≥ 51 g/m(2.7), 33 (21%) subjects were diagnosed with LVH. There was 79% agreement in the diagnosis of LVH between the wall-thickness z-score method and LVMI(2.7) > 51 g/m(2.7) (kappa 0.37). There is poor concordance between the diagnosis of LVH on echocardiogram reports using wall-thickness z-score and diagnosis of LVH using LVMI(2.7) criteria. It is important to establish a consensus method for diagnosing LVH because of the high frequency of cardiovascular complications in children with hypertension.

Comprehensive review on oyster mushroom species (Agaricomycetes): Morphology, nutrition, cultivation and future aspects.

Huge volumes of organic matter are produced on earth via photosynthesis and their disposal is a serious threat to the environment and public health all over the world. Nevertheless, these agricultural wastes possess a chemical composition conducive to mushroom cultivation. Lignocellulosic wastes, comprising cellulose, hemicellulose and lignin, offer vital nutrients for mushroom growth. Oyster mushrooms are well known for their unique ability to degrade lignocellulosic materials, making them valuable contributors to the process of organic waste decomposition and nutrient cycling in ecosystems. Employing agricultural by-products as a substrate for mushroom cultivation presents a sustainable approach to waste reduction and the production of nutritionally enriched food. Cultivating oyster mushrooms, presents an economically feasible and environment friendly method of transforming waste materials into highly nutritious food. These edible mushrooms are widely grown worldwide, comprising around 27 percent of the total global production. Oyster cultivation has rapidly increased in Asia due to its low production technology, easy availability of substrates, temperature tolerance and high yield capacity. Oyster mushrooms are sought after as a functional food due to their appealing taste, aroma, flavor, nutritional benefits and medicinal properties. They contain high levels of protein, fiber, vitamins B complex, C and D2, as well as minerals like potassium, phosphorus, selenium, zinc and essential amino acids. These mushrooms are versatile, as they thrive in both tropical and temperate regions without requiring complex controlled environmental conditions for growth. This review article provides insights into the cultivation aspects of important oyster species including a novel species called Hypsizygus ulmarius. Oyster mushroom cultivation is rapidly growing in developing countries, where it can contribute to food security for the world's growing population, which is expected to reach 9.7 billion by 2050.

Survival in epithelial ovarian cancer: a multivariate analysis incorporating BRCA mutation status and platinum sensitivity.

BACKGROUND: Patients with BRCA-associated ovarian cancer (OC) have a survival advantage over those with sporadic OC. To further explore this, we examined the impact of prognostic factors on disease-free survival (DFS) and overall survival (OS) in patients with known BRCA mutation status. PATIENTS AND METHODS: We reviewed stage III-IV OC patients treated at our institution between 1 December 1996 and 30 September 2006 and also tested on protocol for BRCA mutations. Impact on DFS and OS was determined by Kaplan-Meier analysis and a Cox proportional hazards model. RESULTS: Of the 110 patients, 36 had deleterious BRCA mutations [BRCA (+)] and 74 were BRCA wild type [BRCA(-)]. Thirty-one of 36 (86%) BRCA (+) and 60 of 74 (81%) BRCA (-) patients were platinum sensitive (P = 0.60). Median OS was longer for BRCA (+) patients (not reached versus 67.8 months; P = 0.02), but DFS was similar (26.9 versus 24.0, P = 0.3). On multivariate analysis, OS correlated with primary platinum sensitivity [HR = 0.15; 95% CI (confidence interval) 0.06-0.34] and BRCA (+) mutation status (HR = 0.33; 95% CI 0.12-0.86). CONCLUSIONS: BRCA mutation status predicted OS independent of primary platinum sensitivity, suggesting that underlying tumor biology contributes to disease outcome and may be worthy of consideration in future clinical trial design.

Telmisartan, a dual ARB/partial PPAR-γ agonist, protects myocardium from ischaemic reperfusion injury in experimental diabetes.

AIM: Apart from its angiotensin receptor blocker (ARB) activity, telmisartan is also a partial agonist of peroxisome proliferator-activated receptor gamma (PPAR-γ). Therefore, we assessed whether telmisartan treatment attenuates myocardial ischaemia/reperfusion (I/R) injury in diabetic rats through PPAR-γ pathway. METHODS: Diabetic rats were randomized to receive vehicle (sham and I/R), telmisartan (10 mg/kg/day, orally), PPAR-γ antagonist GW9662 (1 mg/kg/day, intraperitoneally) or both for 14 days. On 15th day, excluding sham group, left anterior descending coronary artery occlusion was performed for 45 min followed by 1 h of reperfusion. Haemodynamic, biochemical, histopathological, ultrastructural, immunohistochemical (Bax and Bcl-2 protein), TUNEL positivity, infarct size and western blot studies were performed. RESULTS: Telmisartan treatment significantly improved cardiac function by normalizing mean arterial pressure, left ventricular pressure (±LVdP/dt(max) , a marker of myocardial contraction and relaxation), by decreasing left ventricular end-diastolic pressure (a marker of preload, 3.7 ± 0.41 vs. 7.3 ± 0.89, p < 0.001) and percent infarct area (37.52 ± 5.83 vs. 46.27 ± 3.20, p < 0.01) as compared to diabetic I/R group. Interestingly, GW9662 worsens the I/R injury (percent infarct area, 54.38 ± 6.48 vs. 46.27 ± 3.20, p < 0.01), whereas telmisartan with GW9662 (percent infarct area, 41.16 ± 8.23 vs. 46.27 ± 3.20, p < 0.05) showed lesser significant results as compared to telmisartan alone. Additionally, telmisartan significantly ameliorates activities of endogenous antioxidants, creatine kinase-MB isoenzyme, lactate dehydrogenase and prevented the increase of tumour necrosis factor-alpha and malondialdehyde in myocardium. Furthermore, telmisartan also decreased Bax expression (4.45 ± 1.24% vs. 10.25 ± 0.96%, p < 0.01), number of TUNEL-positive cells (6.2 ± 0.98% vs. 13.0 ± 1.6, p < 0.01), inflammation, myonecrosis and increased Bcl-2 expression (5.45 ± 0.15% vs. 1.24 ± 0.3%, p < 0.01). On the other hand, GW9662 treatment alone increased the Bax expression, TUNEL positivity and decreased Bcl-2 expression. Telmisartan protective effects were partially attenuated by a co-administration with GW9662. Western blot analysis showed that telmisartan treatment enhanced PPAR-γ expression, whereas GW9662 decreased it in myocardium. CONCLUSIONS: In addition to the class effect of ARBs, telmisartan has a beneficial effect in I/R injury in diabetic rats in part because of activation of PPAR-γ.

Pattern of Fatal Injuries in Counter Terrorist Operations: An Innovative Analysis through Embalming Services.

BACKGROUND: Mortal remains of the soldiers killed in counter-terrorist operations in Kashmir valley are sent to their home after undergoing mandatory embalming. METHODS: Injuries on the mortal remains of the soldiers killed in counter terrorist operations between Jan 1999 to Dec 2006 were analysed with respect to the agent, mode of injury, age, rank structure, body parts involved, seasonal variations and changing trends. Fatalities consequent to enemy action across line of control and fatalities of Kargil war were also analysed for comparison. Statistical analysis was done using chi square test for difference in proportions. RESULT: Over the study period, terrorist induced injuries accounted for 8.16 deaths per thousand troops deployed whereas enemy action from across the line of control accounted for 0.63 deaths per thousand. Terrorist induced fatalities peaked in 2001 and thereafter revealed a declining trend ('p' < 0.001). Fatalities due to enemy action across line of control declined to zero since 25 Nov 2003 consequent to effective ceasefire. Of the total fatalities, 89.5% were killed in action (KIA) while 10.5% died of their wounds after reaching the hospital. Fatality to total injured ratio peaked to 29% in 2001 and than stabilized to about 23%. Mean KIA to total casualty ratio was 21%. The rank structure of the fatalities was officers 8.6%, JCOs 7.3%, and Other Ranks 84.1%. Most of the soldiers died young, 51% being below 25 years of age. Out of the terrorist induced fatalities, 78.2% died of gunshot wounds and 21.5% by splinters and improvised explosive devices (IED). The ratio was reversed in enemy induced fatalities and in Kargil war. Fatalities peaked during June to November and declined in winters. Body region wise, 23.4% of all deaths were due to head injury, 8.4% due to neck and maxillofacial injury, 18.4% due to injury to lungs and 11% due to heart injury. Most frequent target of the fatal bullet was brain (25.4%), closely followed by lungs (22.5%) and heart (12.3%). When soldier died of splinters / IED, multiple body parts were injured in 57.5%, brain in 17.3%, face & neck in 3.5%, heart in 6.6%, lungs in 5.3%, abdomen in 3.5% and limbs in 5.8%. Fatality due to head and heart injury peaked in 2001, while multiple injuries peaked in 2000, declined in 2001 and peaked again in 2004 and 2005 ('p' < 0.001). In fatalities of Kargil war, chest injuries were less but multiple injuries were more. CONCLUSION: Most of the fatalities were due to gunshot wounds selectively aimed at head, face, neck and thorax. Therefore, a lightweight flexible and effective bulletproof protection for this area will conserve manpower.

Feeding strategies in the ELBW infant.

Necrotizing enterocolitis (NEC) remains a common emergency that carries high morbidity and mortality for extremely low birth weight infants. To date there have been no multicenter randomized controlled trials to evaluate different feeding strategies and NEC. Clinicians must weigh their experience against small amounts of data in deciding the best way to feed their patients. Currently published feeding protocols and evidence for the same will be reviewed. Also reviewed is the evidence for use of human milk, initiation and advancement of feedings, and the use of probiotics.

Analysis of Non Enemy Action Deaths in Counter Insurgency Operations through Mortuary Services.

BACKGROUND: Regional Trauma Centre in the northern India receives the mortal remains of all fallen soldiers for embalming. Non enemy action deaths during counter insurgency operations (CI Ops) were analysed for planning preventive measures. METHODS: Mortal remains received for embalming from Jan 1999 to Dec 2006 were analysed with respect to mode of injury, causation, body parts involved, fatality, seasonal variation and changing trends. RESULT: Accidents accounted for 3.02 deaths per thousand troops and environmental factors were responsible for 1.14 deaths per thousand troops deployed. Accidental deaths peaked in 2000, declined in 2001 and then remained more or less static. Of the accidental deaths, 88% were brought in dead and 12% died after reaching hospital. Road traffic accidents were the major killers accounting for 48.2%, followed by accidental discharge of weapon 35.5%. The latter is showing a rising trend from 8% of total accidents in 2001 to 65% in 2005 and 51% in 2006 (p<.01). Most (49.7%) of the deaths were below 25 years of age. Proportion of persons below 25 years was more in fatalities due to accidental gunshot wound. Amongst the road traffic accidents, 40% died of head injury and 51.2% due to multiple injuries. When deaths occurred due to accidental discharge of own weapon, 36.4% had brain injury and 22% heart injury. Of the environmental fatalities all but one were brought in dead. Majority were due to avalanches and landslides (51.2%), followed by earthquake (22.4%), lightning (12.8%), high altitude pulmonary oedema (10.4%) and hypothermia (3.2%). Most of the deaths due to avalanches occurred in February while all deaths due to earthquake were in October 2005. Of the deaths due to lightning, 75% occurred in April and May. CONCLUSION: Prevention of death caused by road traffic accidents, accidental discharge of weapon, avalanches and lightning will conserve manpower and add to operational preparedness.

Splenic Syndrome due to Sickle Cell Trait amongst Indian Soldiers Serving in Kashmir.

BACKGROUND: Heterozygous transmission of gene for Haemoglobin S leads to sickle cell trait. Mostly the trait remains silent with no additional morbidity or mortality. When these persons migrate to higher altitudes, in times of high oxygen demand, some of them develop splenic infarction. This is a rare phenomenon and only 47 such cases had been reported till 2005. METHODS: This study was conducted at an Indian military hospital serving the troops deployed in Kashmir valley at altitudes ranging from 5500 ft to 13000 ft. When two consecutive splenectomies for splenic abscesses, turned out to be sickling induced infarction histopathologically, we reviewed splenectomy specimens received in last six years for evidence of sickling. RESULT: Out of 33 splenectomies performed during the period of study, 22 were due to trauma (gun shot injury 11; splinter injury one and blunt injury 10). Of the rest eleven, who presented without any history of trauma, seven had evidence of vascular occlusion with aggregates of sickled red blood cells. In none, Gram stain or Periodic Acid Schiff stain revealed any bacterial or fungal colonies. One patient of splenic syndrome was found to have unrecognised sickle cell trait and he was managed conservatively. CONCLUSION: Sickle cell trait should be excluded before considering splenectomy in ethnically vulnerable patients presenting with splenic syndrome. An uncomplicated splenic infarction can be managed conservatively.

From laboratory to Phase I/II cancer trials with recombinant biotherapeutics.

Many promising recombinant cancer medicines are generated by academic research and increasing the number of these products that are translated into the clinic will increase the pipeline of new therapies. Recombinant proteins for use in Phase I/II cancer trials must be produced to standards of Good Manufacturing Practice (GMP) in compliance with EU law. This can be a major obstacle for translating experimental products to clinical reality especially when there is no established process or prior experience with GMP. Here, we illustrate the principals of GMP with a step-by-step guide and we show that GMP can be achieved on a relatively small scale in the researchers own institution. The process is exemplified with an antibody-based therapeutic expressed in the yeast Pichia pastoris. The purified product has been used safely in patients and the principles are applicable to any recombinant protein required for Phase I/II cancer trials.

Aggressive Use of Diagnostic Services is Counterproductive.

BACKGROUND: Medical fraternity requisitions diagnostic tests for multiple reasons. More often than not, the tests lead to more tests either to exclude or to confirm doubts raised by the test results. These tests have an inherent morbidity, discomfort and cost. Growing expenditure on diagnostic tests without matching improvement in the health status warrants an internal audit of the laboratory utilization. METHODS: A retrospective utility audit was done for certain routinely advised laboratory tests at a hospital. Blood urea estimation in annual / periodic medical examination (AME/ PME), bleeding and clotting time in pre-anaesthetic check-up and aspartate aminotransferase (AST) and antibodies to hepatitis C virus (anti-HCV) in diagnostic work-up of acute onset jaundice were included in the audit. RESULTS: During the study period, 793 individuals underwent AME / PME and urea estimation did not provide any additional information in these cases which was not inferred by serum creatinine. Similarly, in diagnostic workup of acute onset jaundice, 6049 aspartate aminotransferase (AST) estimations in 1024 patients did not contribute anything more than what was inferred by alanine aminotransferase (ALT). Prevalence of anti HCV antibodies in acute onset jaundice in serving soldiers (11 out of 1225; 0.89%) though more than that in the blood donors from the same population (17 out of 4105; 0.41%) was less than anticipated false positives (18 out of 1225; 1.5%) as per the claimed specificity (98.5%) of the test kit. None of the 2766 bleeding and clotting time tests detected a bleeding or coagulation disorder. CONCLUSION: The study reveals significant overuse of the laboratory that may not be good for the patient and the organization in terms of direct and indirect costs due to false positive results. This laboratory overload adversely affects the quality and availability of laboratory results. Therefore, a test should only be advised, if positive or negative result would dictate a change in patient management.

Reemergence of Chloramphenicol Sensitivity in Enteric Fever.

BACKGROUND: Enteric fever is a global health problem and rapidly developing resistance to various drugs makes the situation more alarming. Drug sensitivity in Salmonella enterica serovar typhi and Salmonella enterica serovar paratyphi A isolated from 45 blood culture positive cases of enteric fever was tested to determine in-vitro susceptibility pattern of prevalent strains in northern India. METHODS: Strains isolated from 45 blood culture positive cases of typhoid and paratyphoid fever over a period of three years were studied and their sensitivity patterns to chloramphenicol, ampicillin, ciprofloxacin, ceftriaxone, nalidixic acid, amikacin and ofloxacin were analysed. RESULTS: Our results show a high sensitivity of both Salmonella enterica serovar typhi (96%) and Salmonella enterica serovar paratyphi A (100%) to chloramphenicol. Sensitivity to ciprofloxacin and amikacin was 88% and 84% respectively. All the isolates were sensitive to ofloxacin, nalidixic acid and ceftriaxone. Sensitivity of Salmonella enterica serovar paratyphi A was 100% to chloramphenicol, ciprofloxacin, ofloxacin, nalidixic acid and ceftriaxone, 95% to amikacin and 30% to ampicillin. Overall 44 out of 45 isolates of Salmonellae were sensitive to chloramphenicol. CONCLUSION: These findings suggest changing pattern of antibiotic resistance in enteric fever with reemergence of chloramphenicol sensitivity in northern India.

Fluid and electrolyte management in the very low birth weight neonate.

Fluid and electrolyte management in the very low birth weight infant is critical to survival. The amount of fluid present in the plasma, interstitial fluid, and cellular fluid changes throughout the fetal and neonatal period, presenting a challenging situation. One of the many factors influencing fluid requirements is the insensible water loss by mechanisms such as evaporation. Low birth weight infants are especially susceptible to this due to their large body surface area and immature skin, often resulting in hypernatremia and the complications associated with it. However, some infants may experience hyperkalemia, hyperglycemia, and/or hyponatremia, resulting in various other complications. Careful monitoring is essential in deciding how to manage these infants. This article aims to discuss the management of fluid and electrolytes in very low birth weight infants and address ways to decrease the morbidity and mortality associated with the imbalances in fluid and electrolytes seen in this population.

Palliative care in the fetus and newborn.

The changing environment in neonatology and perinatology has led to the examination of issues surrounding palliative care. Newborn palliative care should be considered in three general areas: (1) Neonates at the limits of viability. As advances in technology and outcomes become available, it is the responsibility of the health-care community and society to reach a consensus regarding the limits of viability. (2) Neonates with lethal congenital anomalies. When appropriate, and diagnosis and prognosis are certain, why should a family be deprived the opportunity to choose palliative care for the unborn child? (3) Neonates not responsive to aggressive medical management where continuing therapy may prolong suffering and postpone death. The question 'Are you doing for the neonate or to the neonate?' should be asked. These complex issues, along with best interest issues, site, mode and timing of delivery, and the development of palliative care are the subject of this manuscript.

Sodium-dependent high-affinity binding of carnitine to human placental brush border membranes.

The interaction of carnitine with human placental brush-border membrane vesicles was investigated. Carnitine was found to associate with the membrane vesicles in a Na(+)-dependent manner. The time course of this association did not exhibit an overshoot, which is typical of a Na+ gradient-driven transport process. The absolute requirement for Na+ was noticeable whether the association of carnitine with the vesicles was measured with a short time incubation or under equilibrium conditions, indicating Na(+)-dependent binding of carnitine to the human placental brush-border membranes. The binding was saturable and was of a high-affinity type with a dissociation constant of 1.37 +/- 0.03 microM. Anions had little or no influence on the binding process. The binding process was specific for carnitine and its acyl derivatives. Betaine also competed for the binding process, but other structurally related compounds did not. Kinetic analyses revealed that Na+ increased the affinity of the binding process for carnitine and the Na+/carnitine coupling ratio for the binding process was 1. The dissociation constant for the interaction of Na+ with the binding of carnitine was 24 +/- 4 mM. This constitutes the first report on the identification of Na(+)-dependent high-affinity carnitine binding in the plasma membrane of a mammalian cell. Studies with purified rat renal brush-border membrane vesicles demonstrated the presence of Na+ gradient-driven carnitine transport but no Na(+)-dependent carnitine binding in these membrane vesicles. In contrast, purified intestinal brush-border membrane vesicles posses neither Na+ gradient-driven carnitine transport nor Na(+)-dependent carnitine binding.

Preferential adhesion of prostate cancer cells to bone is mediated by binding to bone marrow endothelial cells as compared to extracellular matrix components in vitro.

We have demonstrated previously that the preferential adhesion of prostate cancer cells to human bone marrow endothelial (HBME) cells may contribute to their preferential metastasis to bone. Although a subject of debate, it has been postulated that the endothelial cells of the bone marrow are fenestrated. It is unknown therefore whether prostate cancer cells adhere preferentially to the extracellular matrix (ECM) or the endothelial cells. It has also been demonstrated in other organ systems that the types of cell adhesion molecules on the surface of endothelial cells lining the organ microvasculature are determined, in part, by the ECM of the organ. We investigated how prostate cancer cell adhesion to HBME cells is affected by growing HBME cells on selected organ-derived ECM proteins in vitro. Growth of HBME cells and immortalized human aortic endothelial cells on bone, kidney, and placenta ECM proteins significantly increased their ability to bind PC-3 cells. This increased adhesion was not dose dependent and was not demonstrated with human dermal microvascular endothelial cells. Scanning electron microscopic analysis demonstrated that prostate cancer cells adhered directly to the endothelial cells and not to the underlying substrata. These results suggest that unidentified cell adhesion molecules are expressed or up-regulated on the apical surfaces of human aortic endothelial cells and HBME cells grown on bone, kidney, and placenta ECMs. These results also strongly demonstrate that the adhesion of prostate cancer cells to bone may be initiated by direct binding to endothelial cells rather than direct binding to exposed ECM components.