Haematological and Immunological Parameters in Patients with Chronic Hepatitis B Infection in Zaria, Nigeria.

Hepatitis B virus (HBV) is one of the major causes of morbidity and mortality worldwide. The aim of this study was to determine the haematological and immunological parameters in patients with chronic HBV infection in Zaria, Nigeria. Twenty individuals with confirmed chronic HBV (CHB) infection constituted the subjects while 20 non-HBV-infected individuals were monitored as controls. The subjects were enrolled purposively from the Gastroenterology Clinic of the Ahmadu Bello University Teaching Hospital Shika, Zaria Nigeria. Four millilitres of blood samples were collected from each study participants. Full blood count was conducted using the Swelab Alfa Haematology Analyzer, while CD4+ T-Cell enumeration was performed using the Sysmex Partec CyFlow® Counter IVD flow cytometer according to the manufacturers' instruction. The mean (and standard deviation) age of the 20 participants with CHB was 32.7 (±10.1) years while that of the 20 HBV negative control participants was 30.0 (±7.8) years. Mann-Whitney test showed no significant difference between the two groups in their total WBC (p=0.6634) and granulocytes (p=0.2386). There was a significant increase in the monocytes count (p=0.0151) and a significant decrease in the lymphocytes count (p=0.0006) of patients with CHB compared to the healthy control. There was no significant difference in the mean CD4+ T-lymphocytes count between subjects and controls (p=0.0633). Unpaired Student t-test showed no significant difference between the two groups in the other haematological parameters. This study showed a significant increase in monocytes and decrease in lymphocytes, a phenomenon that characterize the sustenance of infection by immune evasion mechanism.

Effects of elevated CO2 levels on subcellular distribution of trace metals (Cd and Cu) in marine bivalves.

Hypercapnia (elevated CO2 levels) and pollution with trace metals such as Cu and Cd are common stressors in estuarine habitats that can negatively affect physiology and health of marine organisms. Hypercapnia can modulate toxicity of trace metals including Cu and Cd; however, the physiological and cellular mechanisms of the metal-CO2 interactions are not well understood. We investigated the effects of elevated PCO2 (∼800 and 2000µatm) and metal exposure (50µgl-1 of Cu or Cd) on subcellular distribution of metals in two common species of marine bivalves, Eastern oysters Crassostrea virginica and hard shell clams Mercenaria mercenaria. Oysters accumulated higher burdens of Cu and Cd in the gill tissues compared to clams. In both studied species, Cu was predominantly associated with the metabolically active cell compartments (mitochondria, lysosomes, microsomes and cytosolic enzymes), with a modest fraction sequestered by metallothioneins (∼30%) and the insoluble metal-containing granules (MCG) (∼15-20%). Unlike Cu, Cd was largely sequestered by metallothioneins (∼60-70%), with a relatively small fraction associated with the organelles and the cytosolic enzymes. Mitochondria were the main intracellular target for trace metals accumulating higher concentrations of Cd (and in the case of oysters - of Cu) than other organelles or cytosolic enzymes. Cu accumulation in the metabolically active cellular compartments was independent of the CO2 levels, while Cd content of the organelles and cytosolic enzymes increased at elevated PCO2 in both studied species indicating that hypercapnia may enhance cellular toxicity of Cd in bivalves. Hypercapnia suppressed the sequestration capacity of metallothioneins for Cu and Cd in oysters but increased Cu and Cd load in clam metallothioneins. Thus, metal-induced metabolic injury in oysters may be exaggerated by hypercapnia which enhances metal accumulation in the potentially sensitive intracellular fractions and suppresses the metal detoxification capacity. In contrast, clams appear to be more resistant to the combined effects of hypercapnia and metal exposure reflecting more efficient and robust detoxification mechanisms of this species.

Challenging conventional f-element separation chemistry--reversing uranyl(VI)/lanthanide(III) solvent extraction selectivity.

The water soluble tetradentate Schiff base, N,N'-bis(5-sulfonatosalicylidene)-diaminoethane (H2salen-SO3), will readily coordinate to the uranyl(VI) cation, but not to the same extent to trivalent lanthanide cations. This allows for the reversal of conventional solvent extraction properties and opens the possibility for novel separation processes.

Utilisation of sFLC assays - how well do we comply with guidelines?

INTRODUCTION: There is a paucity of data on the utilisation of the serum-free light chain (sFLC) and compliance with published guidelines. METHODS: Serum-free light chain assays requested at our institute for the diagnosis, prognosis and monitoring of plasma cell dyscrasias from July 2008 to March 2010 were compared with the International Myeloma Working Group (IMWG) consensus guidelines. RESULTS: In total, 1150 assays were performed (4.3 assays per patient, range 1-20). Eight hundred and forty-four (73%) of these were performed for multiple myeloma (MM), 188 (16%) for monoclonal gammopathy of undetermined significance, 72 (6%) for AL amyloidosis and 46 (4%) for smouldering myeloma. Of these, 49.6%, 22.9%, 1.4% and 69.6% of the monitoring assays were outside of IMWG recommendations, respectively. Of the 419 assays performed outside of guidelines for MM patients, 404 (96.4%) were due to monitoring of patients with a measurable M protein, while 24 (5.7%) were due to too frequent requesting (≤14 days) with 15 assays (3.6%) being noncompliant on both grounds. CONCLUSION: Utilisation of the sFLC assay shows reasonable adherence to guidelines within our centre. We propose to further optimise usage of the test with the help of administrative processes and education of clinicians.

Acute multifocal placoid pigment epitheliopathy associated with erythema nodosum and a flu-like illness.

Acute multifocal placoid pigment epitheliopathy (AMPPE) is a rare inflammatory vasculitis of the choroid. Despite primarily being a disorder of the eye, a number of extra-ophthalmological features have been described in AMPPE and may accompany the visual disturbances. Such patients may be admitted under the care of physicians for evaluation of a systemic illness. We report AMPPE occurring in a 37-year-old man in his thirties admitted with a flu-like illness, erythema nodosum, visual disturbances and raised inflammatory markers. Conditions such as bacterial endocarditis and vasculitis, were considered before a diagnosis of AMPPE was made. The case illustrates how AMPPE can be a diagnostic dilemma, particularly when associated with extra-ophthalmological features. The epidemiology, clinical features, diagnosis and therapy of AMPPE are also discussed.

Profound lymphopenia and bacteraemia.

Bacteraemia often carries a poor prognosis despite prompt antibiotic therapy and is associated with late morbidity and mortality that is difficult to explain. Here, we describe perisistent B- and T- cell lymphopenia in a cohort of patients with Gram-positive and Gram-negative bacteraemia. This suggests previously unrecognized mechanisms of subversion of immunity by pathogens and might explain the comorbidity of blood stream infection with bacteria.

Structural analysis of the binding modes of minor groove ligands comprised of disubstituted benzenes.

Two-dimensional homonuclear NMR was used to characterize synthetic DNA minor groove-binding ligands in complexes with oligonucleotides containing three different A-T binding sites. The three ligands studied have a C(2) axis of symmetry and have the same general structural motif of a central para-substituted benzene ring flanked by two meta-substituted rings, giving the molecules a crescent shape. As with other ligands of this shape, specificity seems to arise from a tight fit in the narrow minor groove of the preferred A-T-rich sequences. We found that these ligands slide between binding subsites, behavior attributed to the fact that all of the amide protons in the ligand backbone cannot hydrogen bond to the minor groove simultaneously.

Nitrate anaphylaxis.

BACKGROUND: Nitrate and nitrite salts are commonly used to preserve and sustain color in a number of processed meats. To date there have been no described cases of anaphylaxis to either nitrates or nitrites in the literature. OBJECTIVE: We report a patient with anaphylaxis to nitrates and nitrites documented by double-blind, placebo-controlled capsule challenge. METHODS: A 22-year-old previously well male, presented to a tertiary referral center with a 4-year history of recurrent anaphylaxis after eating take-out food. No further episodes occurred while adhering to a strict elimination diet. We performed a double-blind, placebo-controlled capsule challenge both with food substances and starch placebo. These occurred in a hospital setting with full resuscitative procedures available. RESULTS: An acute anaphylactic reaction occurred following a challenge to nitrates and nitrites. Generalized allergic reactions were observed on separate occasions following administration of artificial colorings and metabisulfite. There was a mild urticarial reaction following salicylates. He was placed on a diet free of sulfites, nitrates, nitrites, and low in salicylates and he has had no further reactions. A computer based search of the Medline, Current Contents and EMBASE databases found no previously reported cases of urticaria, angioedema, or anaphylaxis to either nitrates or nitrites. CONCLUSION: The ingestion of processed meats containing nitrate or nitrite salts may be associated with angioedema and anaphylaxis and should be considered when investigating patients with suspected food allergy.

Immunomodulation therapy in colorectal carcinoma.

There has been much progress in the understanding of the relationship between the immune system and colorectal cancer. This has led to the use of immunomodulatory therapy in the adjuvant and palliative treatment of the condition. Although attempts at the use of non-specific immunomodulation with agents such as levamisole, cimetidine, alpha interferon and Bacillus Calmette-Guerin (BCG) have not produced significant clinical benefits when tested in randomized trials in both the adjuvant setting and for metastatic disease, promising results are being obtained with more specific therapy. Edrecolomab [corrected], a murine monoclonal antibody targeting the 17-1A antigen on malignant colorectal cells has produced a reduction in relapse and mortality rates when used as adjuvant treatment following surgery for Dukes' C colon cancer. Active specific therapy with autologous tumour vaccine administered with BCG has produced similar benefits in Dukes' B cancer. Both 3H1 anti-idiotypic antibody against carcinoembryonic antigen and 105AD7 antibody to gp72 glycoprotein have demonstrated in-vitro and in-vivo immune activation against tumour. Non-randomized studies postulate prolongation of survival using these antibodies in advanced disease. These agents are all currently being tested in randomized studies powered to detect meaningful survival differences and clinical benefit. Immune therapy offers the potential of low toxicity therapy in colorectal cancer and may have a role as an adjunct to conventional chemotherapy.

ACL injuries in the skeletally immature patient.

Anterior cruciate ligament injuries are becoming increasingly prevalent in a younger, more athletic population. These injuries require aggressive management given the poor natural history with regard to progressive meniscal damage and advanced degenerative changes. In general, strict nonoperative management has been associated with failure in the individual committed to return to an active lifestyle. As a result a general algorithm can be expressed as follows: 1. Activity limitations and bracing are initially used for the asymptomatic skeletally immature patient with a complete ACL tear. 2. If the patient remains asymptomatic, formal reconstruction can be delayed until skeletal maturity. 3. For the symptomatic patient a precise delineation of their physical development or skeletal age is important. 4. In patients who have reached Tanner IV secondary sexual development or a skeletal age of 13-14 (F/M), a traditional transphyseal reconstruction with hamstring autografts is appropriate. 5. In the skeletally immature patient who has significant skeletal development ahead of them, the judicious use of physeal sparing procedures is an option for the surgeon comfortable with these techniques. In this case the goal is temporization until formal transphyseal reconstruction can be performed after the peak of skeletal growth.

Normalized and proportionalized cemented femoral stem survivorship at 15 years.

We reviewed the clinical and radiographic results of a series of 215 consecutive hip arthroplasties in which a normalized, proportionalized, cemented femoral component was implanted. This component design may encourage more efficient force transmission from prosthesis to cement to bone and lessen hoop stresses and resultant interface failure. A total of 127 hips in 103 patients with a follow-up of 13 to 17 years (average, 14.8 years) were available for evaluation, which included survivorship analysis. An aseptic failure loosening rate of 3.9% at 15 years for this stem favorably compares with other reported series of first-generation and second-generation stems at similar follow-up. Excluding the 8 stems placed in varus, of which 3 required revision (failure rate, 37.5%), the series has a failure rate of 1.6%.

Matrix metalloproteinase inhibitors: applications in oncology.

Matrix metalloproteinases (MMP) are a group of zinc dependent enzymes which include the interstitial collagenases, stromelysins, gelatinases and membrane-type metalloproteinases. They are involved in the remodelling and turnover of the extracellular matrix proteins. They play a role in wound healing and the pathogenesis of arthritis. In malignancies they play a role in tumor invasion, metastasis and angiogenesis. A number of synthetic matrix metalloproteinase inhibitors (MMPIs) have been developed for clinical use. In preclinical tumor models they have shown promising activity in achieving inhibition of MMPs and reducing tumor growth and metastatic spread. Some have also shown additive or synergistic effects with cytotoxic agents. Phase I and II studies in human subjects have defined the main side effects of these agents as being musculoskeletal pains or arthralgias. As they are cytostatic agents rather than cytotoxic in activity conventional measurements of radiological response for assessment are not applicable in trials. Biological activity has been demonstrated in certain cancers by the effects on levels of tumor markers as surrogate markers of tumor response and also by a fibrotic stromal reaction seen in tumor tissue. Newer agents have been developed with selective inhibition of certain MMPs in an attempt to reduce the side effects. A number of phase III human clinical trials evaluating MMPs are being carried out at present but only one has been formally reported so far. This study suggested that marimastat had no survival advantage when compared to chemotherapy with gemcitabine in advanced pancreatic carcinoma. Current trials are assessing efficacy of MMPIs in maintenance of remission after other modalities of therapy or in combination with cytotoxic agents. MMPs have also been demonstrated to play an important role in the articular cartilage destruction seen in both rheumatoid arthritis and osteoarthritis. The use of MMPIs in both ex vivo and in vivo models have shown promising results and trials are in process to assess their potential role in the control of articular destruction. The true therapeutic role of MMPIs await the results of these randomized studies.

Disruption of family rituals as a mediator of the relationship between parental drinking and adult adjustment in offspring.

This study investigated whether disruption of family-of-origin rituals and routines would mediate the relationship between family history of parental problem drinking (FH+, FH-) and two outcomes in adult offspring: adult children of alcoholics (ACOA) personality attributes and drinking problems. The self-reports of 143 students and 129 outpatients indicated that disruption of family routines and rituals mediated the relationship between family history status and both ACOA traits and problem drinking. These results suggest that the mediating effect of family-of-origin rituals and daily routines may explain some of the heterogeneity in adult outcomes of offspring from FH+ families, in both student and clinical samples.

Comparison of the behavioural effects of infusion of carbachol and acetylcholinesterase into the rat substantia nigra.

It has been postulated for many years that acetylcholinesterase (AChE) may play a nonclassical role in the substantia nigra, unrelated to its ability to hydrolyse acetylcholine. In this study the behavioural effects of unilateral infusion of AChE and a cholinergic agonist, carbachol, were compared. Carbachol induced ipsiversive circling over a very short time scale (minutes), whereas AChE induced contraversive circling, but over a longer time course-10 days. Both agents showed selectivity of response within the substantia nigra: acetylcholinesterase was only effective when infused into the most rostral region of the substantia nigra and its effects were limited to the pars compacta. In contrast, carbachol had effects in both the pars compacta and reticulata, with a graded sensitivity to carbachol in the rostral/caudal plane; infusions into rostral regions induced high rates of circling compared to more caudal areas, suggesting that the cholinergic input to the substantia nigra is not homogenous, but greater in rostral regions. This disparity between the effects of carbachol and AChE would, therefore, suggest that AChE is not exerting its long-term behavioural actions via a cholinergic mechanism, both in terms of time course of the response and the areas within the substantia nigra sensitive to these agents.

Disease outcome in patients with low stage renal cell carcinoma treated with nephron sparing or radical surgery.

PURPOSE: We investigated the outcome of nephron sparing surgery in patients with low grade and low stage (Robson stage II or less) renal cell carcinoma. MATERIALS AND METHODS: We retrospectively reviewed the records of 185 patients treated with nephron sparing surgery and 209 matched for patient age and sex, and tumor stage and grade who were treated with radical nephrectomy. Kaplan-Meier survival curves were constructed for progression and survival end points. Multivariate analysis was performed to determine the tumor characteristics independently correlated with progression and cancer death. RESULTS: No significant difference was observed with respect to progression-free, crude or cancer specific survival between the nephron sparing surgery and radical nephrectomy groups. Less than 5% of the patients treated with conservative nephron sparing surgery had local recurrence. Tumor size was a strong independent predictor of outcome, whereas Robson stage was not. Patients treated with radical nephrectomy had a significant cancer specific and progression-free survival advantage when controlling for tumor diameter and grade. However, no difference was observed in patients with primary tumor diameters of 4 cm. or less. CONCLUSIONS: Robson staging is inaccurate in predicting tumor behavior. Patients with tumors larger than 4 cm. and a normal contralateral kidney may be best served by radical nephrectomy rather than elective nephron sparing surgery. However, nephron sparing surgery may result in an outcome similar to that of radical nephrectomy for low grade, low stage renal cell carcinomas of 4 cm. or smaller.

Influence of DNA ploidy and adjuvant treatment on progression and survival in patients with pathologic stage T3 (PT3) prostate cancer after radical retropubic prostatectomy.

OBJECTIVES: To determine whether adjuvant treatment (AT: hormonal or radiation) affects outcome in pathologic Stage T3 (pT3) prostate cancer when analyzed according to DNA ploidy. METHODS: The predictive value of nuclear DNA ploidy and AT on clinical and prostate-specific antigen (PSA) progression and on overall and cause-specific survival after radical retropubic prostatectomy was assessed in 894 patients with pT3 prostate cancer. RESULTS: Mean follow-up was 6.7 years (range, 0.3 to 20). Mean age was 66 years (range, 39 to 79). Six hundred sixty patients (74%) had no immediate AT, 131 (15%) had early adjuvant radiotherapy (ART), and 103 (12%) had early adjuvant orchiectomy (AHT). DNA diploid tumors were found in 445 patients (52%), tetraploid tumors in 346 (41%), and aneuploid tumors in 59 (7%). DNA ploidy was a significant (P < 0.05) prognostic indicator for clinical systemic progression-free survival. With PSA progression (more than 0.2 ng/mL) as an endpoint, ploidy was an even more powerful predictor for outcome (P = 0.004). Use of early AHT or ART was associated with decreased overall clinical progression for diploid and nondiploid tumors (P < 0.001 and P < 0.001, respectively). With respect to PSA progression, ART and AHT were equally effective and superior to no AT only in patients with diploid tumors. However, in patients with nondiploid tumors, only AHT appeared to have improved PSA progression-free survival (P < 0.001) over ART or no AT, which are similar in outcome. CONCLUSIONS: In the present nonrandomized study, AHT was as effective as ART for all endpoints except for PSA more than 0.2 ng/mL progression, for which it appeared to be superior to ART for patients with nondiploid tumors.

Recombinant acetylcholinesterase has behavioural effects in the rat substantia nigra not attributable to its enzymatic activity.

An unexplained action of acetylcholinesterase (AChE) has previously been demonstrated on motor behaviour in the substantia nigra: a single infusion of the protein induced long-term circling behaviour, reflecting sustained increased activity of the nigro-striatal pathway. In this study, a highly purified form of AChE was infused and the long-term behavioural effects were still observed. In addition, recombinant human AChE produced a similar response, suggesting that AChE itself, and not a contaminant, was responsible for these behavioural actions. Butyrylcholinesterase, which also hydrolyses acetylcholine, was without effect. Hence AChE is not exerting these actions by potentiating the neurotransmitter acetylcholine, but via some cholinergic-independent mechanism.

Effect of experimental limbic epilepsy on the estrus cycle and reproductive success in rats.

Epilepsy was induced in female rats by the injection of tetanus toxin (5 mouse LD50) unilaterally into the ventral hippocampus under anesthesia. During the 2-4 weeks that the rats exhibited intermittent spontaneous convulsions, daily vaginal smear tests showed that their estrous cycle was interrupted. In control rats such interruption only occurred for just a few days after the operation. Investigation of mating behavior, on the first night of proestrus, which occurred after 7 weeks from the operation, showed that there were fewer mounts, intromissions or ejaculations from the males, which were caged with previously epileptic animals. These females produced slightly smaller litters than their controls and there was a marked failure of their young to thrive in comparison with those of the control females. This failure appeared to be related to relatively high "stress" levels in the general laboratory environment. The impairment of reproductive success only lasted about 3 months after the original induction of epilepsy since subsequent litters to the same animals thrived normally.