Biomedical subjects

A S Grumach

Publications and source records attributed to A S Grumach.

At least 19 recordsLinked to original sources

PubMed2006

Recurrent infections in partial complement factor I deficiency: evaluation of three generations of a Brazilian family.

We report here on the evaluation of a factor I-deficient Brazilian family (three generations, 39 members) with strong consanguinity. The complete factor I-deficient patients (n = 3) presented recurrent respiratory infections, skin infections and meningitis; one of them died after sepsis. They presented an impaired total haemolytic activity (CH50), low C3, low factor H and undetectable C3dg/C3d. Partial factor I deficiency was detected in 16 family members (normal low cut-off value was 25 microg/ml). Respiratory infections were the most common clinical occurrence among partial factor I-deficient relatives. Two of them were submitted to nephrectomy following recurrent urinary tract infections. An additional two heterozygous relatives presented with arthritis and rheumatic fever. Apparently, patients with partial factor I deficiency are also at higher risk for recurrent infections. Vaccination against capsulated bacteria and the eventual use of prophylactic antibiotics should be considered individually in this patient group.

Adolescent↗

PubMed2004

The use of reverse transcription-PCR for the diagnosis of X-linked chronic granulomatous disease.

Chronic granulomatous disease (CGD) is an inherited disorder of the innate immune system characterized by a defective oxidative burst of phagocytes and subsequent impairment of their microbicidal activity. Mutations in one of the NADPH-oxidase components affect gene expression or function of this system, leading to the phenotype of CGD. Defects in gp91-phox lead to X-linked CGD, responsible for approximately 70% of CGD cases. Investigation of the highly heterogeneous genotype of CGD patients includes mutation analysis, Northern blot or Western blot assays according to the particular case. The aim of the present study was to use reverse transcription (RT)-PCR for the analysis of molecular defects responsible for X-linked CGD in eight Brazilian patients and to assess its potential for broader application to molecular screening in CGD. Total RNA was prepared from Epstein B virus-transformed B-lymphocytes and reverse transcribed using random hexamers. The resulting cDNA was PCR-amplified by specific and overlapping pairs of primers designed to amplify three regions of the gp91-phox gene: exons 1-5, 3-9, and 7-13. This strategy detected defective gp91-phox expression in seven patients. The RT-PCR results matched clinical history, biochemical data (nitroblue tetrazolium or superoxide release assay) and available mutation analysis in four cases. In three additional cases, RT-PCR results matched clinical history and biochemical data. In another case, RT-PCR was normal despite a clinical history compatible with CGD and defective respiratory burst. We conclude that this new application of RT-PCR analysis--a simple, economical and rapid method--was appropriate for screening molecular defects in 7 of 8 X-linked CGD patients.

Child↗

PubMed2004

Immunological analysis in paediatric HIV patients at different stages of the disease.

There are only few clinical studies on complement in well-defined (or characterized) paediatric HIV patients. Aim of this study was to evaluate the complement system and immunoglobulins in HIV-infected children and to correlate data to stage of disease. Blood samples of 127 HIV-infected children (11-134 months; 62 male : 65 female) were collected in order to evaluate humoral immunity. The patients were classified according to CDC clinical (N-asymptomatic; A-mild symptoms such as common recurrent infections; B-moderate symptoms such as Candidiasis and herpes infections, meningitis, sepsis and anaemia; C-severe symptoms such as opportunistic infections and neoplasia) and with respect to immunological criteria (T CD4(+) cell count). Analysis of complement system included the classical (CH50), alternative (APH50) pathway activities and plasma concentrations of mannan-binding lectin (MBL), of the C4 allotypic variants C4A and C4B. (ELISA), and of the C3 split product C3d (rocket immunoeletrophoresis). Immunodiagnosis also included CD4(+) and CD8(+) lymphocyte count and immunoglobulin concentrations. Complement activation and consumption was observed in all patients correlating with disease activity. Activated classical and alternative pathways and elevated C3d were significantly correlated with immunologic category 3. C3d levels were also significantly correlated with immunologic category 1. Undetectable CH50 and APH50 were found in two (group C) and 10 patients (n = 2, A = 2, B = 2, C = 4), respectively. Low MBL values were found in 13/127 but without correlation to disease severity. Undetectable C4B levels were observed in three patients, favouring the diagnosis of a complete deficiency. Although not related to clinical symptomatology, a strong ongoing complement activation can be observed in all stages of HIV infection. In contrast to earlier reports MBL could not be considered as a risk factor for HIV.

Antibodies↗

PubMed1999

Molecular analysis of chronic granulomatous disease caused by defects in gp91-phox.

Chronic granulomatous disease (CGD) is an uncommon inherited disorder of phagocytic cells in which a defective respiratory burst leads to severe recurrent bacterial and fungal infections. The disease is a consequence of mutations in one of the four molecules that constitute the NADPH oxidase system of electron transport, whose most critical component is an unusual flavocytochrome b localized in the plasma and specific granule membranes. Mutations in the CYBB gene (localized in the short arm of the X chromosome) encoding the beta-subunit of this flavocytochrome (gp91-phox), which is are responsible for 60-65% of all cases of CGD. In this paper, we report the molecular characterization of seven unrelated kindreds native from Colombia and Brazil with CGD caused by gp91-phox deficiency. The exons with the possible mutation were identified by single-strand conformational polymorphism (SSCP) of genomic DNA and then confirmed by DNA sequencing. In one patient we found a substitution of A to G in the penultimate nucleotide of intron 12 (IVS12-2A-->G). In four other cases, four different nonsense mutations were detected: R91X, W106X, R157X, and R290X and the other two patients showed missense substitutions: E225V and C244Y. In six of these kindreds, all mothers were carriers but one that did not present any change in the gp91-phox gene, which indicates a de novo mutation in this kindred. Then, these family-specific mutations in gp91-phox produce different structural defects that alter the expression or function of an essential component of phagocyte oxidase.

Adolescent↗

PubMed1997

Complement factor I deficiency in a family with recurrent infections.

Factor I deficiency causes a permanent, uncontrolled activation of the alternative pathway resulting in an increased turnover of C3 and consumption of factor B, factor H and properdin. Factor I deficiency is clinically associated with recurrent bacterial infections already in early infancy, mainly affecting the upper and lower respiratory tract, or presenting as meningitis or septicemia. We here report on a Brazilian family (n = 9) with known consanguinity, where in 3/7 children, suffering from chronic otitis, meningitis, and respiratory infections, a complete factor I deficiency was recognized. One of the patients died after fulminant sepsis. Hemolytic activity of the alternative pathway was not detectable in the patients' sera due to decreased plasma concentrations of C3, factor B and properdin. As a consequence of factor I deficiency, C3b could not be metabolized with the result that no C3-derived split products (C3dg/C3d) were detectable in the patients' sera. In vitro reconstitution with purified factor I restored the regulatory function in the patients' sera with the subsequent cleavage of C3b to C3c and C3dg. Factor H levels were decreased in all patients' sera and found to be tightly complexed with C3b resulting in a modified electrophoretic mobility. Upon factor I reconstitution, factor H was released from C3b regaining its beta 1 electrophoretic mobility. Complement-mediated biological functions like opsonization of bacteria, chemotactic activity and phagocytosis in these patients were impaired. The parents (cousins, 2nd degree) and 3/4 siblings had significantly reduced factor I plasma levels without further alteration in their complement profile. 3 of these obviously heterozygously deficient family members suffered from recurrent bacterial infections of different frequency and severity.

Adolescent↗

PubMed1997

Brazilian report on primary immunodeficiencies in children: 166 cases studied over a follow-up time of 15 years.

One hundred sixty-six cases of primary immunodeficiency diseases (PID) (95 males, 71 females), diagnosed according to WHO criteria, have been registered at the Children's Hospital, University of São Paulo, Brazil. The following frequencies were found: predominantly humoral defects, 60.8% (n = 101); T cell defects, 4.9% (n = 8); combined ID, 9.6% (n = 16); phagocyte disorders, 18.7% (n = 31); and complement deficiencies, 6% (n = 10). IgA deficiency was the most frequent disorder (n = 60), followed by transient hypogammaglobulinemia (n = 14), chronic granulomatous disease (n = 14), and X-linked agammaglobulinemia (n = 9). In comparison to other (national) reports, we observed higher relative frequencies of phagocyte and complement deficiencies. Recurrent infections were the cause of death in 12.7%. Allergic symptoms were observed in 41%, mainly in IgA-deficient, hypogammaglobulinemic, or hyper-IgE patients, and autoimmune disorders in 5%, predominantly in IgA and complement deficiencies. Five patients suffered from BCG dissemination; two of them died. This is the first Brazilian report on PID over an observation time of 15 years.

Adult↗

PubMed1996

Effect of therapy with recombinant human interferon-gamma on the release of nitric oxide by neutrophils and mononuclear cells from patients with chronic granulomatous disease.

The aim of this study was to investigate the effect of recombinant human interferon-gamma (rHuIFN-gamma) therapy on the release of nitric oxide (NO) by neutrophils (NEU) and mononuclear cells (MON) from patients with chronic granulomatous disease (CGD). Five patients with this rare disease received rHuIFN-gamma (50 micrograms/m2 of body surface, given by subcutaneous injection three times a week) for 6 months. Clinical and laboratory evaluations were performed before and after 1 and 6 months of rHuIFN-gamma therapy. Nitric oxide release by NEU and MON was assessed by the ability of these cells to inhibit thrombin-induced washed platelet aggregation. The nitrite (NO2-) and nitrate (NO3-) levels in the supernatant of cultured NEU and MON, as well as in plasma and urine (24 h diuresis), were quantified by high-performance liquid chromatography (HPLC). Conventional immunologic tests for assessing phagocyte and lymphocyte functions and humoral immunity were also performed. Therapy with rHuIFN-gamma for 6 months did not enhance NO synthesis by NEU or MON from the patients with CGD. The urinary but not plasma levels of NO2- and NO3- were elevated after rHuIFN-gamma therapy. Phagocyte and lymphocyte functions as well as humoral immunity were not affected by rHuIFN-gamma therapy. Although few patients were available for the study, we conclude that therapy with rHuIFN-gamma for 6 months did not enhance the synthesis of NO by NEU and MON in CGD patients. Whether the increased excretion of NO2- and NO3- in the urine of CGD patients after rHUIFN-gamma therapy reflects an induction of NO-synthase in cells other than leukocytes remains to be investigated.

Adolescent↗

PubMed1995

Complement and IgG subclasses in agammaglobulinemic patients.

Complement and IgG subclass analysis was performed in six agammaglobulinemic patients suffering from recurrent bacterial infections. Complement analysis included functional activity of the classical (CH50) and the alternative pathway (APH50) of complement, the complement proteins C3, C4, factor B, the regulators C1-inhibitor, factors H and I as well as the C3-derived split product C3dg/C3d. Plasma concentrations of total IgG ranged between < 0.01 and 297 mg/dl. IgM and IgA were almost undetectable. Levels of all IgG subclasses, which in most cases were measurable only by sensitive ELISA, were markedly reduced in all patients. A pronounced activation of the complement system was observed in all patients as revealed by reduced titers of hemolytic function and elevated levels of C3dg/C3d. Despite the fact that complement activation by itself was the major reason for decreased C4 levels, from depressed plasma concentrations of the common allotypic variants C4A and C4B, a heterozygous C4 deficiency could not be excluded in 2/6 patients. Our data clearly show that, by application of sensitive analytical methods, agammaglobulinemic patients vary considerably in their ability to synthesize immunoglobulins. This may, at least in part, explain the heterogeneous pattern of clinical symptoms. The increased susceptibility of agammaglobulinemic patients to bacterial infections is reflected by a highly activated, and thereby depleted complement system.

Adolescent↗

PubMed1993

Neutrophils and mononuclear cells from patients with chronic granulomatous disease release nitric oxide.

1. Chronic granulomatous disease (CGD) is a group of genetic disorders characterised by recurrent severe suppurative infections due to impaired microbial killing. The principal biochemical defect is an impairment in the production of reactive oxygen intermediates by phagocytes. 2. Nitric oxide (NO) is synthesised from the guanidino nitrogen atom(s) of L-arginine and has recently been proposed to be involved in defence mechanisms. The aim of this study was to investigate the involvement of the oxidative burst in the biosynthesis of NO by neutrophils and mononuclear cells from patients with CGD. 3. NO synthesis was assayed by the ability of neutrophils and mononuclear cells to inhibit thrombin-induced washed platelet aggregation while superoxide anion (O2-) production was measured spectrophotometrically by the superoxide dismutase inhibitable reduction of cytochrome c. 4. Neutrophils and mononuclear cells from patients with CGD released NO. This release was inhibited by nitro-L-arginine methyl ester but could be reversed by L-arginine. Zymosan- and PMA-induced O2- production was less than 10% as compared with healthy controls. 5. These results indicate that O2- production is not essential for NO synthesis in human leucocytes.

Arginine↗

PubMed1993

Immunological factors in milk from Brazilian mothers delivering small-for-date term neonates.

Breast milk samples from three groups of Brazilian women were evaluated: G1, mothers delivering term babies of low birth weight (n = 16); G2, mothers delivering preterm babies of appropriate birth weight (n = 20); G3, mothers delivering term babies of appropriate birth weight (n = 30). Milk samples were obtained at 48 h and on the 7th, 15th, 30th and 60th days after delivery and they were analyzed for lysozyme and total IgA levels and for the presence of specific antibodies against Poliovirus types I, II, III, Rotavirus, Herpes simplex virus, Varicella zoster and Cytomegalovirus. The groups were not statistically different in relation to mother's age, parity, type of delivery or socio-economic levels. IgA levels were higher in both low-birth-weight groups (G1 & G2) compared to the control group (G3) throughout the study period. Lysozyme levels decreased up to the 15th day, increasing thereafter up to the 60th day in all groups. Specific antibodies were detected throughout the study period, with no differences among groups. We conclude that breast milk composition of mothers delivering low-birth-weight babies (G1 & G2) was similar despite the different gestational ages.

Adolescent↗

PubMed1993

Nutritional factors in milk from Brazilian mothers delivering small for gestational age neonates.

The composition of breast milk from brazilian mothers delivering low birthweight infants and its adequacy as a source of nutrients for this group has not yet been fully elucidated. A total of 209 milk samples from 66 women were analysed. The mothers were divided into three groups: G1, mothers delivering term babies of low birthweight (TSGA, n = 16); G2, mothers delivering preterm babies of appropriate birthweight (PTAGA, n = 20); G3, mothers delivering term babies of appropriate birthweight (TAGA, n = 30). The following factors were analysed: osmolarity, total proteins and protein fractions, creamatocrit, sodium, potassium, calcium and magnesium. Milk samples were collected 48 h and 7, 15, 30 and 60 days after delivery. The groups did not differ significantly in terms of osmolarity, total proteins and fractions, creamatocrit, calcium, magnesium or potassium throughout the study period. Sodium levels were higher in all samples from mothers of TSGA infants and in samples from mothers of PTAGA infants on the 7th, 15th and 30th days than in milk from the TAGA group. The authors consider the needs of the low birthweight and TAGA infants and that these high sodium levels may be necessary for growth of low birthweight infants.

Brazil↗

PubMed1991

Laboratory screening for the diagnosis of children with primary immunodeficiencies.

To select simple and low-cost laboratory tests that contribute to the diagnosis of primary immunodeficiencies (PID) in children, the medical records of 98 patients with PID were analyzed. White blood cell counts, serum IgG, IgM and IgA determinations, Shick test, isohemagglutinin titers, delayed cutaneous hypersensitivity tests, nitro blue tetrazolium test and hemolytic complement (CH50) determination associated with clinical data led to the diagnosis in 95% of cases. Applying this laboratory screening to patient series studies in other countries, the diagnosis was suggested in at least 92% of the cases. This screening proposal may be used as a guideline in the standardization of complementary tests for the diagnosis of immunodeficiencies in pediatric centers of developing countries and also at the primary medical care level in developed countries.

Clinical Laboratory Techniques↗

PubMed1991

An alternative method for in vitro production of HIV-1-specific antibodies.

We describe the detection of HIV-1-specific antibody secretion by a 24-h culture of peripheral blood mononuclear cells (PBMC) stimulated with disrupted inactivated whole virus adsorbed onto commercial ELISA microwell plates. The method showed high sensitivity and specificity and was capable of accurately detecting HIV-1 infection early after birth (9 of the 19 patients were less than 3 years old) because maternal antibodies do not interfere by giving false-positive results. No false-positive results were obtained with PBMC from 24 HIV-1-seronegative asymptomatic individuals and no false-negative results were obtained for 10 seropositive adult patients (16-46 years of age). This rapid, relatively low cost, simple, highly sensitive and specific assay can be extremely useful for the early diagnosis of pediatric AIDS cases.

Adolescent↗

PubMed1988

Inherited C3 deficiency of the complement system.

1. We report a patient homozygous for C3 deficiency and several heterozygotes from the same family. Upon follow-up, the homozygote was found to suffer several severe bacterial infections, whereas all the heterozygotes were clinically healthy. 2. C3 was undetectable in the homozygous patient, CH50 was very low and factor I was present. Serum capacity to generate chemoattractant stimuli for peripheral leucocytes was similar to that of normal adults as was also observed for one of the heterozygotes. Serum capacity to opsonize yeast was reduced in the presence of autologous and homologous (normal adult) cells. The CH50 levels of heterozygous patients were within the lower range of normality. 3. The parental consanguinity and the homozygosis state observed here are classical signs of recessive autosomal inheritance. However, the lower or below normal C3 levels detected in parents and relatives point to a co-dominant inheritance of gene S with respect to the "null" gene. 4. C3 polymorphism presented a predominantly "slow" pattern in most family members, which, together with the low C3 levels, indicates the expression of S-allotypes.

Antibodies, Anti-Idiotypic↗

PubMed1988

Primary immunodeficiency diseases: a presentation of 6 cases.

The peripheral blood leukocytes of 6 children with clinical data suggestive of primary cellular immunodeficiencies were studied in an attempt to define the cellular basis of these disorders. The phenotype and function of T and B cells were investigated. According to the clinical and laboratory features, the patients were classified as one case of severe combined immunodeficiency (SCID), two of ataxia-telangiectasia (AT), one of Wiskott-Aldrich syndrome (WAS), one of DiGeorge syndrome (DSG), and one of cellular immunodeficiency (CID). The laboratory investigations together with the clinical manifestations permitted a diagnosis of primary immunodeficiency diseases.

B-Lymphocytes↗

PubMedDate not supplied

Detection of influenza, parainfluenza, adenovirus and respiratory syncytial virus during asthma attacks in children older than 2 years old.

BACKGROUND: Viral upper respiratory tract infections (URTI) have been correlated with the onset of asthma attacks in children and viral identification was reported in 14-49 % of nasal samples. The aim of the present study was to detect influenza, parainfluenza, adenovirus and respiratory syncytial virus (RSV) in older children during acute asthma attacks. METHODS: A total of 104 children (2-14 years) were included in four groups: group I: asthmatics with acute attack and URTI; group II: asthmatics without URTI (group I children, 30 days later); group III: non-asthmatics with URTI; group IV: non-asthmatic, asymptomatic children. A diagnosis of URTI was considered when (3 symptoms (cough and/or sneeze, nasal obstruction, hypertrophy of turbinates, pain and/or retropharynx hyperemia, headache and fever) in asthmatics and at least 2 symptoms in non-asthmatics were present, starting within 7 days. Samples of nasal mucosa cells (n = 123) were collected, and culture and indirect immunofluorescence were carried out to identify respiratory syncytial virus, adenovirus, influenza A and B, parainfluenza 1,2 and 3 and rhinovirus. RESULTS: Viral identification rates were higher in the asthmatic groups: 13.9 % in group I, 11.1 % in group II; 2.8 % in group III and 0 in group IV. The following viruses were identified: RSV 2/36, rhinovirus 1/36, adenovirus 1/36 and parainfluenzae 1/36 in group I; adenovirus 2/18 in group II; RSV 1/36 in group III. CONCLUSIONS: The rate of viral identification was higher in asthmatic children, whether symptomatic or not, suggesting a possible susceptibility to viral infections. Virus could also be a triggering factor in attacks, although it is not the most preponderant in older children.

Adenovirus Infections, Human↗

PubMedDate not supplied

[IgA deficiency: clinical and laboratory evaluation of 60 patients from the "Instituto da Criança"].

OBJECTIVES: To evaluate the main clinical findings that suggest DIgA and describe the complications observed. PATIENTS AND METHOD: Sixty IgA deficient patients (IgA < 5mg/dl) were included, retrospectively. They were submitted to clinical evaluation and specific laboratorial tests. Their evolution was followed up to 15 years. RESULTS: The main complaints were: recurrent infections (50%), allergies (34%) or autoimmune diseases (10%). The respiratory system was the most affected by infections, and asthma and rhinitis were the most frequent allergic symptoms. No immunological impairment was detected, except for IgA deficiency. IgG and IgM levels were elevated in 50% of the patients. There was improvement of clinical symptoms and some of the patients became asymptomatic during the follow-up. CONCLUSION: IgA deficiency has a large clinical spectrum and early diagnosis would indicate prophylaxis for infections and allergy.

Adolescent↗

PubMedDate not supplied

Chédiak-Higashi syndrome: presentation of seven cases.

CONTEXT: Chédiak-Higashi Syndrome (CHS) is a rare autosomal recessive disease characterized by recurrent infections, giant cytoplasmic granules, and oculocutaneous albinism. OBJECTIVE: To describe clinical and laboratory findings from CHS patients. DESIGN: Case report. SETTING: The patients were admitted into the Allergy and Immunology Unit of the Instituto da Criança, a tertiary public care institution. CASES REPORT: Seven patients had oculocutaneous albinism, recurrent infections and giant cytoplasmic granules in the leukocytes. One patient had low IgG levels and three showed impaired bactericidal activity of neutrophils. Six patients died of infectious complications during the accelerated phase. Therapy included ascorbic acid and antibiotics. Chemotherapy was used for the accelerated phase in two patients. Bone marrow transplantation (BMT) was proposed for one patient. DISCUSSION: The authors emphasize the need for early diagnosis and therapy of CHS. BMT should be indicated before the accelerated phase of the disease has developed.

Chediak-Higashi Syndrome↗