PD-1 deficiency is not sufficient to induce myeloid mobilization to the brain or alter the inflammatory profile during chronic neurodegeneration.

Innate immune activation is a major driver of neurodegenerative disease and immune regulatory pathways could be potential targets for therapeutic intervention. Recently, Programmed cell death-1 (PD-1) immune checkpoint inhibition has been proposed to mount an IFN-γ-dependent systemic immune response, leading to the recruitment of peripheral myeloid cells to the brain and neuropathological and functional improvements in mice with Alzheimer's disease-like ß-amyloid pathology. Here we investigate the impact of PD-1 deficiency on murine prion disease (ME7 strain), a model of chronic neurodegeneration. Although PD-1 was found to be increased in the brain of prion mice, the absence of PD-1 did not cause myeloid cell infiltration into the brain or major changes in the inflammatory profile. However, we observed a slight exacerbation of the behavioural phenotype of ME7 mice upon PD-1 deficiency. These results do not support the possibility of using immune checkpoint blockade as a therapeutic strategy in neurodegenerative disease.

Effects of natalizumab on oligoclonal bands in the cerebrospinal fluid of multiple sclerosis patients: a longitudinal study.

Retrospective studies show that natalizumab modifies oligoclonal immunoglobulin (IgG) bands (OCBs) in the cerebrospinal fluid (CSF) of multiple sclerosis (MS) patients. In this study, we prospectively analyzed both serum and CSF samples from 24 MS patients, before and after 2 years of natalizumab-based therapy. Our results showed complete (55%) or partial (27%) disappearance of the OCBs in CSF samples that were taken after 2 years of therapy. Intrathecal IgG production, represented by the IgG index and IgGLoc, was also quantitatively reduced. Our data showed that natalizumab substantially modulates both intrathecal polyclonal and oligoclonal IgG production: This effect was much more potent than was previously reported.

Glycomolecule modifications in the seminiferous epithelial cells and in the acrosome of post-testicular spermatozoa in the alpaca.

A lectin histochemical investigation of the seminiferous epithelium and acrosomes of spermatozoa present in the efferent ductules and epididymal regions was carried out in the alpaca. The histochemical characterization was performed using a battery of different lectins: Con-A, UEA-I, LTA, WGA, GSA-IB4, SBA, PNA, ECA, DBA, MAL-II and SNA. Sialidase digestion and deglycosilation pre-treatments were also employed. The cytoplasm of the Sertoli cells contained N-linked oligosaccharides with α-D-Man/α-D-Glc and GlcNAc and O-linked glycans with α-L-Fuc, ß-GalNAc, ß-D-Gal-(1-4)-D-GlcNAc, α-Gal and Neu5Acα2,6α-GalNAc moieties whereas ß-D-Gal-(1-3)-D-GalNAc residues were included in both O- and N-glycoproteins. Spermatogonia expressed α-D-Man/α-D-Glc residues included in N-glycoproteins and α-Fuc in O-glycoproteins. Spermatocytes contained the N-glycoproteins residues α-D-Man/α-D-Glc and GlcNAc and the O-glycoproteins residues α-L-Fuc, ß-D-Gal-(1-4)-D-GlcNAc, α-Gal, ß-GalNAc, Neu5Acα2,6α-GalNAc and Neu5Acα2,6ß-D-Gal-(1-3)-D-GalNAc. The results of the present study show differences in the presence and distribution of lectin reactive sites throughout the acrosomal development in the alpaca. In particular, Fuc moieties were found only during the Golgi-phase of spermatids, α-Gal were found in the acrosome of Golgi- and cap-phase spermatids, sialic-acid/α-GalNAc sequence was revealed during the cap-phase and elongated spermatids, and α-D-Man/α-D-Glc and GlcNAc were detected only in the acrosomes of elongated spermatids. Finally, ß-GalNAc, ß-D-Gal-(1-3)-D-GalNAc and ß-D-Gal-(1-4)-D-GlcNAc were added to acrosomal glycoproteins in the early stages of spermatogenesis and remained unchanged during the later phases. Differences in the carbohydrate expression were also demonstrated on the sperm acrosomes during passage through the post-testicular ducts.

Artemisinin-type drugs for the treatment of hematological malignancies.

Qinghaosu, known as artemisinin (ARS), has been for over two millennia, one of the most common herbs prescribed in traditional Chinese medicine (TCM). ARS was developed as an antimalarial drug and currently belongs to the established standard treatments of malaria as a combination therapy worldwide. In addition to the antimalarial bioactivity of ARS, anticancer activities have been shown both in vitro and in vivo. Like other natural products, ARS acts in a multi-specific manner also against hematological malignancies. The chemical structure of ARS is a sesquiterpene lactone, which contains an endoperoxide bridge essential for activity. The main mechanism of action of ARS and its derivatives (artesunate, dihydroartemisinin, artemether) toward leukemia, multiple myeloma, and lymphoma cells comprises oxidative stress response, inhibition of proliferation, induction of various types of cell death as apoptosis, autophagy, ferroptosis, inhibition of angiogenesis, and signal transducers, as NF-κB, MYC, amongst others. Therefore, new pharmaceutically active compounds, dimers, trimers, and hybrid molecules, could enhance the existing therapeutic alternatives in combating hematologic malignancies. Owing to the high potency and good tolerance without side effects of ARS-type drugs, combination therapies with standard chemotherapies could be applied in the future after further clinical trials in hematological malignancies.

Expression of prostate glycoconjugates in the stallion and castrated horse.

This work was undertaken to determine the glycoconjugates secreted by the epithelium of the prostate in the intact stallion and castrated horse using lectin histochemical procedures in conjunction with enzymatic digestion and deglycosylation treatments. Additionally, anti-5 and 13-16-cytokeratin antibodies were used to localize epithelial basal cells. In the stallion, lectin histochemistry showed the following sugar residues in the Golgi zone of the glandular cells: α-Glu/Man, α-Fuc and ß-Gal included in both O- and N-linked oligosaccharides as well as ß-GalNAc, GlcNAc and α-Gal, which belonged to O-glycoproteins. ß-Gal and ß-GalNAc moieties were also noted subterminal to sialyl residues. Sialic acid specific lectins identified Neu-5Ac(α2,3-6)-ß-Gal or Neu5Ac(α2,6)-ß-GalNAc sequences in both N- and O-bound glycoproteins. The prostatic glandular cells of the castrated horse expressed some of the same sugar moieties found in the stallions, such as α-Glu/Man, α-Gal and GlcNAc, but significant differences were also noted. In particular, ß-D-GalNAc was only detected subterminal to sialic acid, ß-D-Gal-(1-3)-D-GalNAc was found in N-linked glycans, whereas ß-D-Gal-(1-4)-D-GlcNAc and Neu5Acα2,6Gal/GalNAc were noted only in O-glycoproteins. These results indicate that the lectin binding patterns in glandular cells may be modified by sex hormones. No specific lectin labelling of basal cells was found in either the stallion or the castrated horse even though they were immunostained with specific anti-cytokeratin antibodies. These cells stained more strongly in the castrated horse than in the intact stallion suggesting that they are androgen responsive. The glycomolecules detected in the equine prostate secretions may contribute to the remodelling of the sperm surface, which occurs during sperm transit through the male genital tract and also after ejaculation in the seminal plasma. These changes may be important in the understanding of the stallion fertility.

Day-restricted feeding during pregnancy and lactation programs glucose intolerance and impaired insulin secretion in male rat offspring.

AIM: The maternal environment during pregnancy and lactation plays a determining role in programming energy metabolism in offspring. Among a myriad of maternal factors, disruptions in the light/dark cycle during pregnancy can program glucose intolerance in offspring. Out-of-phase feeding has recently been reported to influence metabolism in adult humans and rodents; however, it is not known whether this environmental factor impacts offspring metabolism when applied during pregnancy and lactation. This study aims to determine whether maternal day-restricted feeding (DF) influences energy metabolism in offspring. METHODS: Pregnant and lactating Wistar rats were subjected to ad libitum (AL) or DF during pregnancy and lactation. The offspring born to the AL and DF dams were intra- and interfostered, which resulted in 4 group types. RESULTS: The male offspring born to and breastfed by the DF dams (DF/DF off) were glucose intolerant, but without parallel insulin resistance as adults. Experiments with isolated pancreatic islets demonstrated that the male DF/DF off rats had reduced insulin secretion with no parallel disruption in calcium handling. However, this reduction in insulin secretion was accompanied by increased miRNA-29a and miRNA34a expression and decreased syntaxin 1a protein levels. CONCLUSION: We conclude that out-of-phase feeding during pregnancy and lactation can lead to glucose intolerance in male offspring, which is caused by a disruption in insulin secretion capacity. This metabolic programming is possibly caused by mechanisms dependent on miRNA modulation of syntaxin 1a.

Human T-cell lymphotropic virus tax and Epstein-Barr virus DNA in peripheral blood of multiple sclerosis patients during acute attack.

OBJECTIVES: A study was performed to determine whether persistent or latent viruses are reactivated during the acute attack in relapsing remitting multiple sclerosis (MS). MATERIAL AND METHODS: DNA of herpes simplex virus type 1 and 2 (HSV-1 and -2), human cytomegalovirus (HCMV), Epstein-Barr virus (EBV), JC virus (JCV) and HTLV-I was searched, using nested polymerase chain reaction (PCR), in peripheral blood mononuclear cells (PBMCs) collected from 14 MS patients on the first day and, twice a week, during an acute attack of the disease. RESULTS: Viral DNA was detected, in at least one PBMC sample, in all the patients. Interestingly, EBV DNA was found in 42.8% of the patients on the first day, while a sharp increase of the HTLV tax-rex DNA frequency (35.7%) was observed on the tenth day. CONCLUSIONS: In MS relapse EBV DNA detection is an early, frequent event, while the finding of tax-rex, but not of other HTLV-I genomic regions, is a secondary phenomenon, suggesting that these two factors could interact in the pathogenesis of MS relapses.

Synthesis and antiviral activity of certain 9-beta-D-ribofuranosylpurine-6-carboxamides.

To examine the structural parameters necessary for antiviral efficacy of certain purine nucleosides, several 9-beta-D-ribofuranosylpurine-6-carboxamides have been synthesized. Glycosylation of the Me3Si derivative of purine--6-carboxamide with protected ribofuranose in the presence of a Lewis acid gave the blocked nucleoside which on deprotection furnished 9-beta-D-ribofuranosyl-6-iodopurine with cyanide ion. Certain 2-amino- and 2-methyl-9-beta-D-ribofuranosylpurine-6-carboxamides have also been prepared. 8-Carbamoylguanosine (16) has been prepared by homolytic acylation of the parent nucleoside. These compounds were tested against several RNA and DNA viruses in cell culture. 9-beta-D-Ribofuranosylpurine-6-carboxamide (6a), the corresponding 6-thiocarboxamide (7b), and 4-amino-8-(beta-D-ribofuranosylamino)pyrimido[5,4-d]pyrimidine (8) showed significant in vitro antiviral activity at nontoxic dosage levels. 6a employed in the treatment of Rift Valley fever virus infected mice at 50 (mg/kg)/day gave a 55% survival rate on day 21 compared to a 30% survival in the controls.

HTLV tax-rex DNA and antibodies in idiopathic amyotrophic lateral sclerosis.

In an attempt to verify the possible role of retrovirus in idiopathic amyotrophic lateral sclerosis (ALS), the sera of 21 ALS patients admitted to the Neurological Unit of the Don Gnocchi Foundation in Milan, Italy, and of 9 ALS patients from Ulm University in Germany have been evaluated for the presence of antibodies to the human T-lymphotropic viruses (HTLV-I and HTLV-II). The sera of 30 healthy individuals and 20 HIV-infected but HTLV-negative subjects have been also studied as control. Moreover, the HTLV tax-rex and pol DNA sequence have been searched in the peripheral blood mononuclear cells (PBMCs) of 15 ALS patients and 15 HIV-positive HTLV-negative subjects using a nested PCR currently employed in our laboratory for the study of HTLV infections. Antibodies to one or more HTLV proteins have been found by using a Western blot (WB) kit in the sera of 10 Italian and 7 German ALS cases, while all the healthy controls were negative and only one HIV-positive subject had antibodies to HTLV gp21. HTLV tax-rex sequences have been found in the PBMCs of 6 ALS patients while all the controls were negative. All 15 ALS cases and controls were negative for HTLV pol DNA indicating that only the most conserved region of the HTLV genome could be detected. On the whole our data indicate that some ALS patients have antibodies to HTLV proteins and that the tax-rex region of the HTLV genome can be found in their PBMCs.

Transmission line analysis of noncylindrical birdcage resonators.

Transmission line theory, validated for the standard cylindrical birdcage coil, has been employed for the analysis of a rectangular birdcage resonator which is useful for MR imaging of the hand. Due to lack of cylindrical symmetry in the rectangular coil, RF field uniformity was evaluated and found to be critically dependent upon the choice of column from which the coil was linearly driven. Effective L1 and L2 inductance elements were determined using known formulas for self and mutual inductance contributions, and compensation of the different inductance elements of the hand coil was performed to produce cylindrically symmetric birdcage current patterns. RF field mapping using the Biot-Savart law demonstrated a rectangular coil sensitivity 22% greater than a comparable cylindrical version. MR hand and wrist images were acquired using the rectangular birdcage coil. In addition, transmission line analysis was extended to other noncylindrical birdcage geometries.

Use of transmission line analysis for multi-tuning of birdcage resonators.

A general analysis for double- and higher order tuning of birdcage resonators is presented based on a lumped element transmission line model. Expressions were developed for the determination of the resonant frequencies of bandstop and bandpass birdcage coils and, with specific restrictions, for capacitor values required to obtain any two desired mode one resonant frequencies. Experimental measurements on three variants each of an eight-column bandstop and an eight-column bandpass were in excellent agreement with theory; the average absolute frequency difference and percent deviation were 1.51 +/- 1.57 MHz and 2.61 +/- 2.36%, respectively. In addition, 31P and 1H phantom images were obtained at 2.0 T using a representative bandstop coil.

Laryngomalacia. The search for the second lesion.

OBJECTIVES: To determine the necessity of rigid endoscopy in the diagnosis and management of laryngomalacia and its associated synchronous airway lesions (SALs), to analyze the incidence of SALs associated with laryngomalacia and their significance, and to determine the need for epiglottoplasty in management of laryngomalacia. DESIGN: Retrospective medical chart review. SETTING: Tertiary care children's hospital. PATIENTS: Two hundred thirty-three patients with a primary diagnosis of laryngomalacia on flexible fiberoptic laryngoscopy treated at the Children's National Medical Center, Washington, DC, from January 1, 1984, to June 30, 1994. INTERVENTION: Evaluation and treatment of laryngomalacia and associated SAL by flexible fiberoptic laryngoscopy, radiographic studies, rigid endoscopy, and other surgical procedures. MAIN OUTCOME MEASURES: Resolution of airway symptoms from laryngomalacia and associated SAL. RESULTS: Ninety patients (38.6%) underwent rigid endoscopy, and 12 patients (5.2%) required epiglottoplasty. Synchronous airway lesions were discovered in 44 patients (18.9%). Eleven patients (4.7%) had SALs that wre considered clinically significant; nine (3.9%) of these required surgical intervention. CONCLUSIONS: Rigid endoscopy in evaluation of an infant with laryngomalacia is rarely necessary. Clinically significant SALs requiring surgical intervention are uncommon. Surgical intervention for laryngomalacia also is rarely necessary.

Henoch-Schönlein syndrome associated with human Parvovirus B19 primary infection.

We report a case of Henoch-Schönlein syndrome associated with human Parvovirus B19 primary infection. The patient, a 27-year-old Filipino woman, presented with an erythemato-papular-purpuric eruption localized to the lower limbs. General symptoms and signs included fever, hypotension, nausea, vomiting, abdominal pain, inguinal lymphadenopathy and polyarthralgia. Laboratory examinations showed leukocytosis, increase in total serum IgA, proteinuria and haematuria. Circulating IgA immune complexes were also present. The ELISA test for anti-human Parvovirus B19 IgM was positive. Histopathological examination revealed a leukocytoclastic vasculitis. This case confirms that also in adult patients, Henoch-Schönlein syndrome may be associated with human Parvovirus B19 infection.

Stridor in neonates.

Stridor in neonates and infants is a symptom that indicates partial obstruction of the large diameter airways. Its presence should prompt a thorough examination and workup. Steps in evaluating stridor include a careful history and physical examination and rapid assessment of the severity of the clinical situation. Infants with respiratory distress and severe stridor should be safely and urgently transported to a tertiary care center, and colleagues from the departments of otolaryngology and anesthesia-critical care should be alerted. An essential component of the physical examination is auscultation. The phase of respiration in which the stridor is heard best provides important clues to help localize its cause. Radiographs, including plain films, dynamic fluoroscopic airway films, contrast esophagography, CT, and MR imaging are useful in specific clinical situations, based on the likely differential diagnosis. The anatomic causes for stridor in infants and neonates are vast. Successful management depends on expert consultation, proper equipment, and a staff that is experienced in the management of pediatric airway problems. The trend over the past decade has been to significantly decrease morbidity and mortality and also to decrease the number of tracheotomies necessary to stabilize pediatric airways. The best treatment outcomes result when there is good cooperation and communication among pediatricians, otolaryngologists, pulmonologists, and anesthesiologists.

Molecular analysis of HTLV-I and HTLV-II isolates from Italian blood donors, intravenous drug users and prisoners.

Using molecular methods three or five major variants of HTLV-I have been identified; moreover two subtypes of HTLV-II defined as HTLV-IIa and HTLV-IIb with six variants within each of these groups have been described. In the present study we analysed proviral DNA obtained from the peripheral blood mononuclear cells (PBMCs) of a significant group of Italian intravenous drug users (IVDUs), prison inmates and blood donors (BDs) who were HTLV antibody positive. Restriction fragment length polymorphism (RFLP) of amplified LTR region with ApaI, NdeI, DraI, SacI and MaeIII endonucleases was used to define the HTLV-I subtypes, while the different variants of HTLV-IIa and -IIb were defined by RFLP of the LTR region with the AvaII, BglI, SauI, XhoI and BanII endonucleases. The four HTLV-I isolated from BDs were characterized as C type. All the 11 HTLV-II detected in the IVDUs were HTLV-IIb4, while among the prisoners one HTLV-IIb5 and five HTLV-IIb4 were found. Interestingly, in the BDs group two HTLV-IIa0 and one HTLV-IIb4 were detected. It should also be noted that 82% of the IVDUs and 50% of the prisoners were coinfected with HIV, while all the BDs were HIV negative. These data indicate that HTLV-IIb4 is the predominant genotype in Italian IVDUs and prisoners, while the significant variability observed in the BD HTLV-II isolates could be due to the different source of infection among this group.

Immunological patterns during regular intensive training in athletes: quantification and evaluation of a preventive pharmacological approach.

The effects of regular intensive exercise training on immune system homeostasis and the potential value of treatment with an immunostimulating agent were assessed in this randomized, double-blind, placebo-controlled, parallel-group study. A total of 60 athletes were studied over a 3-month period of regular intensive physical activity. After 1 and 3 months there were significant decreases in the immunoglobulin levels in the whole athlete population compared with baseline values. Specifically there were significant decreases in immunoglobulin M and immunoglobulin G (G1 and G2 subclasses). There was also a significant decrease in natural killer cells and a slight but significant increase in B and T lymphocytes. In the thymomodulin-treated group, unlike the placebo group, there was no significant decrease in the immunoglobulin G2 subclass and there was a significant increase in the T-helper cell subpopulation. The clinical relevance of these immunological findings should be evaluated in larger clinical and epidemiological studies.

Estimating the true costs of disposable and reusable instruments.

The real issue in the debate about reusable versus disposable surgical instruments is the true cost and efficacy of specific instruments. Each device needs to be scrutinized on its own merits and analytically compared to other instruments. The goal of this case study was not to promote one type of instrument over the other but to focus on some of the data elements and complex issues involved in determining true costs, cost-effectiveness, and efficacy of the reusable and disposable instruments.

Entomofauna of a buried body: study of the exhumation of a human cadaver in Buenos Aires, Argentina.

This study focuses on insects and other arthropods sampled on the exhumation of an infant skeleton belonging to 'Prof. Dr. Rómulo Lambre' skeletal collection. The body was buried in soil inside a wooden coffin in a grave 40cm deep, in autumn, and stored in the cemetery deposit after exhumation. Death records were obtained from the cemetery archive. Samples of faunal remains were recovered from wrappings, clothes, bones and soil samples, and were identified at different taxonomic levels depending on the stage of conservation. The dominant taxon was the muscid fly Ophyra aenescens (Wiedemann). The relationships among the identified taxa and the moving of the corpse, from the burial context to the cemetery deposit, are discussed and used to create a hypothetical colonization sequence after death. The application of entomological data to anthropological research can provide valuable information for the interpretation of taphonomic processes and burial contexts.

Bromide ion exchange with a Keggin polyoxometalate on functionalized polymeric membranes: a theoretical and experimental study.

Noncovalent interactions between the polyoxometalate [PMo12O40](3-) and acryloyloxyundecyltrimethyl ammonium bromide surfactant, used during membrane preparation, were evaluated in the frame of density functional theory. The electronic solvation energy of [PMo12O40](3) and bromide anions was also evaluated, at the same level of theory, in order to predict a probable exchange on the polymeric surface between these anions at the water/polymer interface. Energy balances were theoretically assessed, showing that the bromide cannot be exchanged with this nanosized polyanion in large extent. In order to validate this theoretical conclusion, ad hoc and accurate measurements were carried out by using homemade polymeric membranes and by dipping them in an ca. 0.4 mM solution of Na3[PMo12O40] for 4 days. The Br(-) concentration, released in a polyoxometalate solution, was followed at different times during the test period by gravimetrical analysis. The agreement between the theoretical prediction and experimental data was remarkable, as the quantum calculations correctly accounted for the short-range intermolecular interactions involved in this phenomenon. Bearing in mind that the achieved conclusion is based on an ab initio quantum approach, the findings of this study can be considered rather general and then exploitable for other similar systems.

Continuous exposure to Kaposi sarcoma-associated herpesvirus (KSHV) in healthcare workers does not result in KSHV infection.

Kaposi sarcoma (KS)-associated herpesvirus (KSHV or HHV-8) infection routes and risk of occupational exposure are still ill-defined. We analysed the risk for occupational acquisition of KSHV infection in healthcare workers (HCWs) with prolonged professional exposure to patients with classic KS, comparing the results to those obtained in healthy relatives of KS patients. Serum and/or saliva KSHV-specific antibodies and DNA were detected in five out of 35 healthy relatives of KS patients but in none of the eight HCWs, suggesting that, outside strict family contacts, horizontal transmission of KSHV is highly inefficient even for HCWs with prolonged contact with KS patients.