Toxoplasma gondii in lactating animals: potential risk to milk consuming population in Khyber Pakhtunkhwa.

Toxoplasma gondii is an intracellular zoonotic protozoan parasite usually infects human and animal worldwide. This study aimed to analyze the sero-prevalence of T. gondii in blood of lactating animals and human living in close proximity and also to detect Toxoplasma DNA in unpasteurized milk of the studied animals. A total of 233 blood and milk samples were collected from lactating animals, and 735 blood samples were taken from humans in District Upper Dir, Khyber Pakhtunkhwa, Pakistan. The blood samples were analyzed through ELISA while the milk samples were analyzed by PCR for the presence of T. gondii DNA. A standard questionnaire was introduced to collect the data from the participants. In animals, the reported sero-prevalence was 32.18% for IgM, 17.16% for IgG, and 6.4% for both IgM and IgG. The reported positivity for T. gondii DNA in milk was 14.44%, 34.8%, 20%, and 26% in sheep, goats, cows, and buffaloes, respectively. In the human blood samples, 9.8% were found positive for IgM and 11.2% for IgG while none of the samples was found positive for both IgM and IgG. Overall sero-prevalence reported in females was significantly higher than the male (p<0.05) poor hygiene condition (p < 0.0001) were the significant risk factors associated with T. gondii infections in animals. In conclusion, T. gondii infection is prevalent in lactating animals and humans using their raw milk in the study area. It is suggested that raw milk should be considered as a vehicle for the transmission of T. gondii to humans. Proper pasteurization of milk is very useful in limiting the transmission of infection. Awareness and control programs should be implemented to prevent the infection.

Human-black bear conflict: crop raiding by Asiatic black bear (Ursus thibetanus) in Azad Jammu and Kashmir, Pakistan.

Asiatic black bear has long been in conflict with human beings crop raiding is a major cause of this conflict frequently noted in South Asia. Crops raided by black bears affected by temporal, spatial and anthropogenic attributes. Insight in this conflict and its mitigation is vital for the conservation of this threatened species. Present study aimed to evaluate crop raiding by black bears in the mountainous region of Azad Jammu and Kashmir. Field surveys were carried out to observe spatial and temporal crop raiding features between 2015-2020 and data gathered using designed questionnaires randomly tailored in villages nearby the forests. Results revealed that maize was the sole crop raided by black bears. A total of 28-acre area was raided by black bear in the fall season (Aug-November) resulting in a damage of 51 metric tons, whole raiding was carried out at night. Each respondent received crop damage on 0.09 acre with a loss of 0.17 metric ton yield. Crop quantity and area were significantly correlated to each other. District Neelum shared 49% of the total crop loss, while 47% of the maize was raided at the altitudinal range of 2100-2500 m. crop raiding was highly significantly ( χ 2 = 1174.64 ; d f = 308 ;   p < 0.01 ) dependent upon distance to the forest. Linear regression revealed that maize quantity was determined by area, time and the total field area. Farmers faced 3.8 million PKRs loss due to crop damage by black bears. Despite the huge loss, the majority (23%) of the respondents did not respond to the query on mitigation measures indicating a poor adaptation of preventive measures. Preferred strategy to avoid crop damage was making noise (27.8%) when bears attacked their crops. A start of compensation scheme to the farmers is recommended that will have turned their negative attitude into a positive one toward the wildlife and black bear particularly. Study provides a new insight in human-bear conflict, particularly in spatial and temporal context of crop raiding in AJ&K.

Rydberg energies and transition probabilities of Li I for np-ms (m ≤ 5) transitions.

Background: Mathematical modeling provides grounds for understanding scientific systems theoretically. It serves as a guide for experimentalists in determining directions of investigation. Recently, the Covid-19 pandemic has caused disturbances in almost every walk of life. Scientists have played their role and have continued research on the effects of the pandemic. Various mathematical models have been used in different branches of science (Djilali et al. in Phys Scr 96 12 124016, 2021; Math Biosci Eng 18(6):8245-8256, 2021; Zeb et al. in Alex Eng J 61(7):5649-5665). Well-established mathematical models give results close to those obtained by experiments. The Weakest Bound Electron Potential Model is one such model, which explains hydrogen-like atoms and ions. This model has been used extensively for hydrogen-like atoms and ions to calculate energies of Rydberg levels and ionization energies. This model has been used extensively for hydrogen-like atoms and ions to calculate energies of Rydberg levels and ionization energies. Results: This paper presents the energies of the Rydberg series, 2s2ns, and 2s2np of Li I, calculated using WBEPM. The energies are used to calculate transition probabilities from np to 2s, 3s, 4s, and 5s levels. The transition probabilities are compared with corresponding values in published data where available. The agreement with known values is good; most of the transition probabilities calculated in this work are new. A computer program was developed to find the value of the dipole matrix element. The calculations were further verified by calculating the lifetimes of some low-lying levels. Conclusions: Four series of Li I have been studied, and energies of the Rydberg levels in the series were calculated. The energies then are used to calculate transition probabilities from np to ms transitions, where m = 2, 3, 4, & 5 and n = 1-15. The results are compared where available. An excellent agreement with previously published data shows the reliability of calculations. Most of the transition probabilities are new.

Food habits of indian crested porcupine (Hystrix indica) (Kerr 1792), in district Bagh, Azad Jammu and Kashmir / Hábitos alimentares do porco-espinho de crista indiano (Hystrix indica) (Kerr 1972) nos distritos de Bagh, Azad Jammu e Caxemira

The Indian Crested Porcupine (Hystrix indica) is classified as an agricultural pest species. It feeds on plants and crops; hence, it is responsible for massive financial losses worldwide. The current study was conducted to assess the diet composition of Indian Crested Porcupine in District Bagh, Azad Jammu and Kashmir (AJ&K). Thus, fecal samples were collected and examined from different sampling sites. Reference slides of the material collected from the study area were prepared for identification of dietary components in fecal pellets. A total of 80 fecal samples were collected and processed. Percent relative frequencies (P.R.F.) were calculated for each plant species recovered from pellets. Data revealed that Indian Crested Porcupine consumed 31 plant species in its diet, among them Zea mays (34.31±7.76) was the most frequently selected species followed by Rumex obtusifolius (15.32±2.57) and Melia azedarach (12.83±4.79). The study revealed that the greatest diversity of (n=20) plant species were consumed in summer season while minimum (n=13) species were used during winter. Among the parts of plants, stem was highly consumed in spring (57.2%) as compared to seed in fall (36.7%) while spikes and leaf were the least recovered parts from the fecal matter. The Berger-Parker diversity index showed highly diversified food (10.92) in the summer time of the year as compared to the autumn season (2.95). This study provides a baseline for the diet preference of this pest in the study area. Based on current findings, a detailed investigation on damage assessment, exploration, habitat use and management of Indian Crested Porcupine in AJ&K has been recommended.

O porco-espinho de crista indiano (Hystrix indica) é classificado como uma espécie de praga agrícola. Alimenta-se de plantas e colheitas; portanto, é responsável por enormes perdas financeiras em todo o mundo. O estudo atual foi realizado para avaliar a composição da dieta do porco-espinho de crista indiano nos distritos de Bagh, Azad Jammu e Caxemira (AJ&K). Assim, amostras fecais foram coletadas e examinadas em diferentes locais de amostragem. Lâminas de referência do material coletado na área de estudo foram preparadas para identificação dos componentes da dieta em pellets fecais. Um total de 80 amostras fecais foi coletado e processado. As frequências relativas percentuais (P.R.F.) foram calculadas para cada espécie de planta recuperada de pelotas. Os dados revelaram que o porco-espinho indiano consumiu 31 espécies de plantas em sua dieta, entre elas Zea mays (34,31 ± 7,76) foi a espécie mais selecionada, seguida por Rumex obtusifolius (15,32 ± 2,57) e Melia azedarach (12,83 ± 4,79). O estudo revelou que a maior diversidade de (n = 20) espécies de plantas foi consumida no verão, enquanto o mínimo (n = 13) espécies foi utilizado durante o inverno. Entre as partes das plantas, o caule foi muito consumido na primavera (57,2%) em relação à semente no outono (36,7%), enquanto a espiga e a folha foram as partes menos recuperadas da matéria fecal., O índice de diversidade de Berger-Parker mostrou alimentos altamente diversificados (10,92) no verão do ano em comparação com o outono (2,95). Este estudo fornece uma linha de base para a preferência alimentar dessa praga na área de estudo. Com base nas descobertas atuais, uma investigação detalhada sobre avaliação de danos, exploração, uso de habitat e gerenciamento de porco-espinho de crista indiano em AJ&K foi recomendada.

Food habits of indian crested porcupine (Hystrix indica) (Kerr 1792), in district Bagh, Azad Jammu and Kashmir

Abstract The Indian Crested Porcupine (Hystrix indica) is classified as an agricultural pest species. It feeds on plants and crops; hence, it is responsible for massive financial losses worldwide. The current study was conducted to assess the diet composition of Indian Crested Porcupine in District Bagh, Azad Jammu and Kashmir (AJ&K). Thus, fecal samples were collected and examined from different sampling sites. Reference slides of the material collected from the study area were prepared for identification of dietary components in fecal pellets. A total of 80 fecal samples were collected and processed. Percent relative frequencies (P.R.F.) were calculated for each plant species recovered from pellets. Data revealed that Indian Crested Porcupine consumed 31 plant species in its diet, among them Zea mays (34.31±7.76) was the most frequently selected species followed by Rumex obtusifolius (15.32±2.57) and Melia azedarach (12.83±4.79). The study revealed that the greatest diversity of (n=20) plant species were consumed in summer season while minimum (n=13) species were used during winter. Among the parts of plants, stem was highly consumed in spring (57.2%) as compared to seed in fall (36.7%) while spikes and leaf were the least recovered parts from the fecal matter. The Berger-Parker diversity index showed highly diversified food (10.92) in the summer time of the year as compared to the autumn season (2.95). This study provides a baseline for the diet preference of this pest in the study area. Based on current findings, a detailed investigation on damage assessment, exploration, habitat use and management of Indian Crested Porcupine in AJ&K has been recommended.

Resumo O porco-espinho de crista indiano (Hystrix indica) é classificado como uma espécie de praga agrícola. Alimenta-se de plantas e colheitas; portanto, é responsável por enormes perdas financeiras em todo o mundo. O estudo atual foi realizado para avaliar a composição da dieta do porco-espinho de crista indiano nos distritos de Bagh, Azad Jammu e Caxemira (AJ&K). Assim, amostras fecais foram coletadas e examinadas em diferentes locais de amostragem. Lâminas de referência do material coletado na área de estudo foram preparadas para identificação dos componentes da dieta em pellets fecais. Um total de 80 amostras fecais foi coletado e processado. As frequências relativas percentuais (P.R.F.) foram calculadas para cada espécie de planta recuperada de pelotas. Os dados revelaram que o porco-espinho indiano consumiu 31 espécies de plantas em sua dieta, entre elas Zea mays (34,31 ± 7,76) foi a espécie mais selecionada, seguida por Rumex obtusifolius (15,32 ± 2,57) e Melia azedarach (12,83 ± 4,79). O estudo revelou que a maior diversidade de (n = 20) espécies de plantas foi consumida no verão, enquanto o mínimo (n = 13) espécies foi utilizado durante o inverno. Entre as partes das plantas, o caule foi muito consumido na primavera (57,2%) em relação à semente no outono (36,7%), enquanto a espiga e a folha foram as partes menos recuperadas da matéria fecal., O índice de diversidade de Berger-Parker mostrou alimentos altamente diversificados (10,92) no verão do ano em comparação com o outono (2,95). Este estudo fornece uma linha de base para a preferência alimentar dessa praga na área de estudo. Com base nas descobertas atuais, uma investigação detalhada sobre avaliação de danos, exploração, uso de habitat e gerenciamento de porco-espinho de crista indiano em AJ&K foi recomendada.

Food habits of indian crested porcupine (Hystrix indica) (Kerr 1792), in district Bagh, Azad Jammu and Kashmir / Hábitos alimentares do porco-espinho de crista indiano (Hystrix indica) (Kerr 1972) nos distritos de Bagh, Azad Jammu e Caxemira

The Indian Crested Porcupine (Hystrix indica) is classified as an agricultural pest species. It feeds on plants and crops; hence, it is responsible for massive financial losses worldwide. The current study was conducted to assess the diet composition of Indian Crested Porcupine in District Bagh, Azad Jammu and Kashmir (AJ&K). Thus, fecal samples were collected and examined from different sampling sites. Reference slides of the material collected from the study area were prepared for identification of dietary components in fecal pellets. A total of 80 fecal samples were collected and processed. Percent relative frequencies (P.R.F.) were calculated for each plant species recovered from pellets. Data revealed that Indian Crested Porcupine consumed 31 plant species in its diet, among them Zea mays (34.31±7.76) was the most frequently selected species followed by Rumex obtusifolius (15.32±2.57) and Melia azedarach (12.83±4.79). The study revealed that the greatest diversity of (n=20) plant species were consumed in summer season while minimum (n=13) species were used during winter. Among the parts of plants, stem was highly consumed in spring (57.2%) as compared to seed in fall (36.7%) while spikes and leaf were the least recovered parts from the fecal matter. The Berger-Parker diversity index showed highly diversified food (10.92) in the summer time of the year as compared to the autumn season (2.95). This study provides a baseline for the diet preference of this pest in the study area. Based on current findings, a detailed investigation on damage assessment, exploration, habitat use and management of Indian Crested Porcupine in AJ&K has been recommended.

O porco-espinho de crista indiano (Hystrix indica) é classificado como uma espécie de praga agrícola. Alimenta-se de plantas e colheitas; portanto, é responsável por enormes perdas financeiras em todo o mundo. O estudo atual foi realizado para avaliar a composição da dieta do porco-espinho de crista indiano nos distritos de Bagh, Azad Jammu e Caxemira (AJ&K). Assim, amostras fecais foram coletadas e examinadas em diferentes locais de amostragem. Lâminas de referência do material coletado na área de estudo foram preparadas para identificação dos componentes da dieta em pellets fecais. Um total de 80 amostras fecais foi coletado e processado. As frequências relativas percentuais (P.R.F.) foram calculadas para cada espécie de planta recuperada de pelotas. Os dados revelaram que o porco-espinho indiano consumiu 31 espécies de plantas em sua dieta, entre elas Zea mays (34,31 ± 7,76) foi a espécie mais selecionada, seguida por Rumex obtusifolius (15,32 ± 2,57) e Melia azedarach (12,83 ± 4,79). O estudo revelou que a maior diversidade de (n = 20) espécies de plantas foi consumida no verão, enquanto o mínimo (n = 13) espécies foi utilizado durante o inverno. Entre as partes das plantas, o caule foi muito consumido na primavera (57,2%) em relação à semente no outono (36,7%), enquanto a espiga e a folha foram as partes menos recuperadas da matéria fecal., O índice de diversidade de Berger-Parker mostrou alimentos altamente diversificados (10,92) no verão do ano em comparação com o outono (2,95). Este estudo fornece uma linha de base para a preferência alimentar dessa praga na área de estudo. Com base nas descobertas atuais, uma investigação detalhada sobre avaliação de danos, exploração, uso de habitat e gerenciamento de porco-espinho de crista indiano em AJ&K foi recomendada.

Food habits of indian crested porcupine (Hystrix indica) (Kerr 1792), in district Bagh, Azad Jammu and Kashmir.

The Indian Crested Porcupine (Hystrix indica) is classified as an agricultural pest species. It feeds on plants and crops; hence, it is responsible for massive financial losses worldwide. The current study was conducted to assess the diet composition of Indian Crested Porcupine in District Bagh, Azad Jammu and Kashmir (AJ&K). Thus, fecal samples were collected and examined from different sampling sites. Reference slides of the material collected from the study area were prepared for identification of dietary components in fecal pellets. A total of 80 fecal samples were collected and processed. Percent relative frequencies (P.R.F.) were calculated for each plant species recovered from pellets. Data revealed that Indian Crested Porcupine consumed 31 plant species in its diet, among them Zea mays (34.31±7.76) was the most frequently selected species followed by Rumex obtusifolius (15.32±2.57) and Melia azedarach (12.83±4.79). The study revealed that the greatest diversity of (n=20) plant species were consumed in summer season while minimum (n=13) species were used during winter. Among the parts of plants, stem was highly consumed in spring (57.2%) as compared to seed in fall (36.7%) while spikes and leaf were the least recovered parts from the fecal matter. The Berger-Parker diversity index showed highly diversified food (10.92) in the summer time of the year as compared to the autumn season (2.95). This study provides a baseline for the diet preference of this pest in the study area. Based on current findings, a detailed investigation on damage assessment, exploration, habitat use and management of Indian Crested Porcupine in AJ&K has been recommended.

Cannabidiol Ameliorates Cognitive Function via Regulation of IL-33 and TREM2 Upregulation in a Murine Model of Alzheimer's Disease.

There is a dire need for due innovative therapeutic modalities to improve outcomes of AD patients. In this study, we tested whether cannabidiol (CBD) improves outcomes in a translational model of familial AD and to investigate if CBD regulates interleukin (IL)-33 and triggering receptor expressed on myeloid cells 2 (TREM2), which are associated with improved cognitive function. CBD was administered to 5xFAD mice, which recapitulate early onset, familial AD. Behavioral tests and immunoassays were used to evaluate cognitive and motor outcomes. Our findings suggest that CBD treatment enhanced IL-33 and TREM2 expression, ameliorated the symptoms of AD, and retarded cognitive decline.

Towards athermal organic-inorganic guided mode resonance filters.

We demonstrate guided-mode resonance filters featuring an amorphous TiO(2) layer fabricated by atomic layer deposition on a polymeric substrate. The thermal properties of such filters are studied in detail by taking into account both thermal expansion of the structure and thermo-optic coefficients of the materials. We show both theoretically and experimentally that these two effects partially compensate for each other, leading to nearly athermal devices. The wavelength shift of the resonance reflectance peak (< 1 nm) is a small fraction of the peak width (~11 nm) up to temperatures exceeding the room temperature by tens of degrees centigrade.

Sprouty 2 binds ESCRT-II factor Eap20 and facilitates HIV-1 gag release.

The four ESCRT (endocytic sorting complexes required for transport) complexes (ESCRT-0, -I, -II, and -III) normally operate sequentially in the trafficking of cellular cargo. HIV-1 Gag trafficking and release as virus-like particles (VLPs) require the participation of ESCRTs; however, its use of ESCRTs is selective and nonsequential. Specifically, Gag trafficking to release sites on the plasma membrane does not require ESCRT-0 or -II. It is known that a bypass of ESCRT-0 is achieved by the direct linkage of the ESCRT-I component, Tsg101, to the primary L domain motif (PTAP) in Gag and that bypass of ESCRT-II is achieved by the linkage of Gag to ESCRT-III through the adaptor protein Alix. However, the mechanism by which Gag suppresses the interaction of bound ESCRT-I with ESCRT-II is unknown. Here we show (i) that VLP release requires the steady-state level of Sprouty 2 (Spry2) in COS-1 cells, (ii) that Spry2 binds the ESCRT-II component Eap20, (iii) that binding Eap20 permits Spry2 to disrupt ESCRT-I interaction with ESCRT-II, and (iv) that coexpression of Gag with a Spry2 fragment that binds Eap20 increases VLP release. Spry2 also facilitated release of P7L-Gag (i.e., release in the absence of Tsg101 binding). In this case, rescue required the secondary L domain (YPX(n)L) in HIV-1 Gag that binds Alix and the region in Spry2 that binds Eap20. The results identify Spry2 as a novel cellular factor that facilitates release driven by the primary and secondary HIV-1 Gag L domains.

Mapping of a new SGBS locus to chromosome Xp22 in a family with a severe form of Simpson-Golabi-Behmel syndrome.

Simpson-Golabi-Behmel syndrome (SGBS) is an X-linked overgrowth syndrome with associated visceral and skeletal abnormalities. Alterations in the glypican-3 gene (GPC3), which is located on Xq26, have been implicated in the etiology of relatively milder cases of this disorder. Not all individuals with SGBS have demonstrated disruptions of the GPC3 locus, which raises the possibility that other loci on the X chromosome could be responsible for some cases of this syndrome. We have previously described a large family with a severe form of SGBS that is characterized by multiple anomalies, hydrops fetalis, and death within the first 8 wk of life. Using 25 simple tandem-repeat polymorphism markers spanning the X chromosome, we have localized the gene for this disorder to an approximately 6-Mb region of Xp22, with a maximum LOD score of 3.31 and with LOD scores <-2.0 for all of Xq. These results demonstrate that neither the GPC3 gene nor other genes on Xq26 are responsible for all cases of SGBS and that a second SGBS locus resides on Xp22.

Purification and characterization of hemolysin from Porphyromonas gingivalis A7436.

Porphyromonas gingivalis, a periodontal pathogen, has the ability to lyse erythrocytes. The hemolytic activity of P. gingivalis A7436 was purified as a 45-kDa protein from the culture supernatant of a 3-days old culture using nickel-nitrilotriacetic acid chromatography. Erythrocytes treated with purified P. gingivalis hemolysin showed the presence of pores and extracellular debris by scanning electron microscopy. Active immunization of mice with 15 micrograms hemolysin induced neutralizing antibodies to hemolysin. Heating at 60 degrees C and treatment with trypsin and dithiothreitol abolished hemolytic activity, while incubation with the protease inhibitor Na-p-tosyl-L-lysine chloromethyl ketone caused no effect. We report here for the first time purification of a hemolysin from P. gingivalis A7436. The amino acid sequence of an internal peptide of hemolysin showed sequence similarity with fimbrillin from P. gingivalis HG564. However, the amino acid composition of purified hemolysin was different from that of P. gingivalis fimbrillin. Also, the ability to lyse but not agglutinate erythrocytes and to bind to nickel-nitrilotriacetic acid differentiates P. gingivalis hemolysin from fimbrillin.

Invasion of aortic and heart endothelial cells by Porphyromonas gingivalis.

Invasion of host cells is believed to be an important strategy utilized by a number of pathogens, which affords them protection from the host immune system. The connective tissues of the periodontium are extremely well vascularized, which allows invading microorganisms, such as the periodontal pathogen Porphyromonas gingivalis, to readily enter the bloodstream. However, the ability of P. gingivalis to actively invade endothelial cells has not been previously examined. In this study, we demonstrate that P. gingivalis can invade bovine and human endothelial cells as assessed by an antibiotic protection assay and by transmission and scanning electron microscopy. P. gingivalis A7436 was demonstrated to adhere to and to invade fetal bovine heart endothelial cells (FBHEC), bovine aortic endothelial cells (BAEC), and human umbilical vein endothelial cells (HUVEC). Invasion efficiencies of 0.1, 0.2, and 0. 3% were obtained with BAEC, HUVEC, and FBHEC, respectively. Invasion of FBHEC and BAEC by P. gingivalis A7436 assessed by electron microscopy revealed the formation of microvillus-like extensions around adherent bacteria followed by the engulfment of the pathogen within vacuoles. Invasion of BAEC by P. gingivalis A7436 was inhibited by cytochalasin D, nocodazole, staurosporine, protease inhibitors, and sodium azide, indicating that cytoskeletal rearrangements, protein phosphorylation, energy metabolism, and P. gingivalis proteases are essential for invasion. In contrast, addition of rifampin, nalidixic acid, and chloramphenicol had little effect on invasion, indicating that bacterial RNA, DNA, and de novo protein synthesis are not required for P. gingivalis invasion of endothelial cells. Likewise de novo protein synthesis by endothelial cells was not required for invasion by P. gingivalis. P. gingivalis 381 was demonstrated to adhere to and to invade BAEC (0.11 and 0.1% efficiency, respectively). However, adherence and invasion of the corresponding fimA mutant DPG3, which lacks the major fimbriae, was not detected. These results indicate that P. gingivalis can actively invade endothelial cells and that fimbriae are required for this process. P. gingivalis invasion of endothelial cells may represent another strategy utilized by this pathogen to thwart the host immune response.

Haemolysin from Mycobacterium avium complex isolates from AIDS patients.

Cell-bound haemolytic activity was observed in isolates of Mycobacterium avium complex (MAC) from AIDS patients. M. avium type strains showed negligible activity. None of the culture supernates exhibited any haemolytic activity. Zwitterionic detergent 3-[(3-cholamidopropyl)-dimethyl-ammonio]-1-propanesulphonate (CHAPS) was used to extract haemolysin from ethanol-treated M. avium complex strain 101 (MAC101) cells. Haemolysin was isolated from CHAPS extract (CE) by metal affinity chromatography and identified as a 32-kDa protein by polyclonal antibodies raised against M. tuberculosis haemolysin. Treatment of CE with trypsin resulted in reduction of haemolytic activity, whereas heating at 100 degrees C for 10 min did not affect its activity. A similar 32-kDa haemolysin was extracted from cells of M. avium K128 which was isolated from a monkey infected with simian immunodeficiency virus (SIV). The haemolysin produced by M. avium strains isolated from AIDS patients may be associated with the pathogenesis of M. avium infection.

Isolation of a contact-dependent haemolysin from Mycobacterium tuberculosis.

Contact-dependent haemolytic activity was observed with cells of Mycobacterium tuberculosis H(37)Rv and M. tuberculosis H(37)Ra, but not with those of M. bovis, M. bovis BCG and M. africanum. Culture filtrates of all these strains did not exhibit any haemolytic activity. M. tuberculosis H(37)Rv was subsequently used for the isolation of haemolysin. Haemolytic activity was retained in the cell debris even after sonication of the cells and treatment with Tween 80 and lysozyme. Solubilisation of haemolysin was possible only after the cell debris was washed with ethanol 70% and then treated with Tween 80 0.1%. The haemolysin thus obtained showed a micellar M(r) of >200000 by gel-filtration on Sephadex G-200 and a subunit M(r) of 66000 by SDS-PAGE. It was sensitive to trypsin but stable when heated at 60 degrees C for 10 min. Polyclonal serum raised in rabbits against the haemolysin neutralised the haemolytic activity. The N-terminal amino-acid sequence of the 66-kDa subunit of haemolysin showed identity with TB66, the 66-kDa secretory protein of M. tuberculosis, and 30% homology with the haemolysin A precursor of Vibrio cholerae. Phosphatidylglycerol inhibited lysis of sheep erythrocytes by the haemolysin and is probably the receptor for the haemolysin. Haemolysin not only lysed erythrocytes, but was also cytotoxic to human lung cells. It appears that, among the members of the M. tuberculosis complex, the cell-bound contact-dependent haemolysin/cytolysin is restricted to M. tuberculosis and it may be associated with the pathogenesis of M. tuberculosis.

Isolation of a 33-kDa protein antigen from delipidified Mycobacterium tuberculosis H37Rv.

A 33-kDa protein (TB33) was isolated from a delipidated cell sonicate (CS) of Mycobacterium tuberculosis H37Rv (grown in Middlebrook 7H9 broth supplemented with glucose) using immobilized metal affinity chromatography (IMAC) on a nickel-nitrilotriacetic acid (Ni-NTA) column. TB33 could not be isolated from the culture filtrate (CF) of M. tuberculosis H37Rv using Ni-NTA. TB33 was recognized by monoclonal antibodies (mAb) known to react with proteins of M. tuberculosis with a molecular mass of 33/34 kDa; namely, mAb F126-5, F67-1 and F126-2. The N-terminal amino acid sequence of TB33 was found to be Xaa-Xaa-Thr-Pro-Ala-Asp-Val-Ser/Cys-Asn-Val-Ala-Ile and thus, shows identity with the N-terminal of antigen 84 of M. tuberculosis except for two mismatches. Antibodies to TB33 could be raised in mice by administering four injections of TB33 (40 micrograms total protein). Sera from tuberculosis patients reacted with TB33, while those from normal healthy individuals did not.

Immunoaffinity chromatographic isolation of a high molecular weight seroreactive protein from Mycobacterium leprae cell sonicate.

The purpose of this study was to isolate Mycobacterium leprae antigen(s) by immunoaffinity chromatography using immunoglobulins from leprosy patients and from rabbit anti-M. leprae hyperimmune serum coupled to CNBr-Sepharose 4B. A high molecular weight (M(r)) M. leprae protein (MLP) with a subunit M(r) of 22,000 was isolated. MLP was recognized by monoclonal antibody MMPII1G4 which is known to react with MMPII, a 22 kDa protein of M. leprae. The N-terminal sequence of the 22 kDa subunit (Met-gln-gly-asp-pro-asp-val-leu-arg-leu-leu-asn-glu-gln-leu-thr) was identical to MMPII and to antigen D (bacterioferritin) of M. paratuberculosis. It showed 44% homology with N-terminal end of E. coli bacterioferritin. In ELISA, MLP showed 100% and 60% positivity with leprosy and TB sera respectively as compared to normal healthy sera. The role of bacterioferritin in M. leprae and the importance of MLP as an immunogen has been discussed.

Perinatal mortality in the Cape Province, 1989-1991.

OBJECTIVE: To determine the number of deliveries, the low-birth-weight rate and the perinatal mortality rate at provincial and province-aided hospitals and clinics in each planning region of the Cape Province. DESIGN: A record of the number of deliveries, low-birth-weight infants, stillbirths and early neonatal deaths in provincial and province-aided hospitals and clinics in each planning region of the Cape Province between 1 January 1989 and 31 December 1991. SETTING: All provincial and province-aided hospitals and clinics in the Cape Province. PARTICIPANTS: Hospital and clinic staff of all provincial and province-aided hospitals and clinics in the Cape Province. MAIN OUTCOMES MEASURED: Number of deliveries, low-birth-weight infants, stillbirths and early neonatal deaths in each planning region of the Cape Province. RESULTS: A total of 373,768 births were recorded during the 3-year period with a low-birth-weight rate of 14.7%, a stillbirth rate of 17.9 per 1,000 and an early neonatal mortality rate of 9.1 per 1,000. All rates differed widely between regions. The regions with the highest perinatal mortality rates were in the northern and eastern Cape. CONCLUSION: The perinatal demographics of most regions in the Cape Province are typical of a developing country. Regions which have been identified as having the highest low-birth-weight, stillbirth and early neonatal mortality rates are in greatest need of improved perinatal health care.

Isolation of a 43 kDa protein from Mycobacterium tuberculosis H37Rv and its identification as a pyridine nucleotide transhydrogenase.

A 43 kDa protein (TB43) was isolated from the cell sonicate (CS) of Mycobacterium tuberculosis H37Rv with immobilized metal affinity chromatography (IMAC) on a Ni-nitrilotriacetic acid column. Two-dimensional electrophoresis of the IMAC fraction showed a major spot with an M(r) of 43,000 and a pI of approximately 6.0. The N-terminal amino acid sequence of TB43 was met-arg-val-gly-ile-pro-asn-glu-thr-lys-asn-asn-glu-phe-arg-val-ala- ile-thr-pro-ala. It showed 86% homology with the N-terminal end of the alanine dehydrogenase of Myco. tuberculosis and 65% homology with the N-terminal end of the alpha-subunit of the Escherichia coli pyridine nucleotide transhydrogenase (Tsh). TB43 did not show any alanine dehydrogenase activity and did not react with monoclonal antibody (MAb) HBT10, which is known to recognize the 40 kDa alanine dehydrogenase of Myco. tuberculosis. It was also not recognized by MAb F29-29 which is known to react with a 43 kDa protein of Myco. tuberculosis complex. This protein exhibited strong Tsh activity. A similar 43 kDa protein showing Tsh activity was also isolated by IMAC from Myco. bovis CS. However, the pI of the protein was approximately 7.0. A similar protein could not be isolated from the CS or culture filtrate of Myco. bovis BCG and Myco. tuberculosis H37Ra. TB43 is a cell-associated pyridine nucleotide transhydrogenase and is distinct from the 40/44 kDa secreted alanine dehydrogenase of Myco. tuberculosis.

Skin reactivity and fibronectin-binding property of TB66 (66-kDa protein of Mycobacterium tuberculosis).

A 66-kDa protein (TB66) was purified from culture filtrate (CF) and cell sonicate (CS) of Mycobacterium tuberculosis H37Rv by immobilised metal affinity chromatography (IMAC) on a Ni-nitrilotriacetic acid (NTA) column. TB66 was found to be a fibronectin-binding protein as determined by ELISA and could be purified by affinity chromatography with fibronectin-Sepharose. A similar 66-kDa protein could be isolated also from M. bovis, M. bovis BCG, M. africanum and M. tuberculosis H37Ra by IMAC, but not from any other mycobacteria. The NH2-terminal amino-acid sequence of TB66 from H37Rv and M. bovis was identical and showed 85% homology with the N-terminal sequence of bovine serum albumin (BSA). A monoclonal antibody (MAb) OD4AG3 recognised a heat-stable and trypsin-sensitive epitope near the C-terminal end of TB66. This MAb also recognised the 66-kDa protein isolated from the other members of the M. tuberculosis complex. In tests of immunogenicity, TB66 elicited a delayed type hypersensitivity reaction in guinea-pigs immunised with either TB66 or with M. tuberculosis H37Rv. TB66 also elicited an antibody response in immunised guinea-pigs and stimulated murine macrophages to produce tumour necrosis factor.